Journal articles: 'Indirect ELISA'

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Kohl, Thomas O., and Carl A. Ascoli. "Indirect Immunometric ELISA." Cold Spring Harbor Protocols 2017, no. 5 (May 2017): pdb.prot093708. http://dx.doi.org/10.1101/pdb.prot093708.

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Nasim, Faiz-ul-Hassan, Samina Ejaz, Muhammad Ashraf, and Gulzar Ahmad. "Indirect Back-Titration ELISA." Applied Immunohistochemistry & Molecular Morphology 24, no. 1 (January 2016): 64–70. http://dx.doi.org/10.1097/pai.0000000000000150.

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Morenkov, O. S., Nadja Fodor, and I. Fodor. "Indirect Elisas Based on Recombinant and Affinity-Purified Glycoprotein E of Aujeszky's Disease Virus to Differentiate Between Vaccinated and Infected Animals." Acta Veterinaria Hungarica 47, no. 1 (January 1, 1999): 137–50. http://dx.doi.org/10.1556/avet.47.1999.1.15.

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Two indirect ELISAs for the detection of antibodies against glycoprotein E (gE) of Aujeszky's disease virus (ADV) in sera have been developed. The rec-gE-ELISA is based on theE. coli-expressed recombinant protein containing the N-terminal sequences of gE (aa 1-125) fused with the glutathione S-transferase fromSchistosoma japonicum. The affi-gE-ELISA is based on native gE, which was purified from virions by affinity chromatography. The tests were optimised and compared with each other, as well as with the recently developed blocking gE-ELISA (Morenkov et al., 1997b), with respect to specificity and sensitivity. The rec-gE-ELISA was less sensitive in detecting ADV-infected animals than the affi-gE-ELISA (sensitivity 80% and 97%, respectively), which is probably due to the lack of conformation-dependent immunodominant epitopes on the recombinant protein expressed inE. coli. The specificity of the rec-gE-ELISA and affi-gE-ELISA was rather moderate (90% and 94%, respectively) because it was necessary to set such cut-off values in the tests that provided a maximum level of sensitivity, which obviously increased the incidence of false positive reactions. Though the indirect ELISAs detect antibodies against many epitopes of gE, the blocking gE-ELISA, which detects antibodies against only one immunodominant epitope of gE, showed a better test performance (specificity 99% and sensitivity 98%). This is most probably due to rather high dilutions of the sera used in the indirect gE-ELISAs (1:30) as compared to the serum dilution in the blocking gE-ELISA (1:2). We conclude that the indirect gE-ELISAs are sufficiently specific and sensitive to distinguish ADV-infected swine from those vaccinated with gE-negative vaccine and can be useful, in particularly affi-gE-ELISA, as additional tests for the detection of antibodies to gE.

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Wreghitt, T. G., and Margaret Sillis. "A μ-capture ELISA for detecting Mycoplasma pneumoniae IgM: comparison with indirect immunofluorescence and indirect ELISA." Journal of Hygiene 94, no. 2 (April 1985): 217–27. http://dx.doi.org/10.1017/s0022172400061428.

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SUMMARYA μ-capture ELISA was developed for detecting Mycoplasma pneumoniae specific IgM, and compared with an indirect immunofluorescent antibody (IFA) technique and an indirect ELISA. μ-capture ELISA and IFA compared well and were found to be the most sensitive assays. The IFA test can be completed in 2 h whilst the results of the μ-capture ELISA can be available in 24 h. Both tests are amenable to routine diagnostic use and have similar sensitivity. Indirect ELISA was found to be less sensitive and less specific, giving high assay values with several sera having undetectable M. pneumoniae CF antibody or CF antibody in low titre. Serum samples obtained from 11 patients at various times after M. pneumoniae infection showed maximum antibody levels within the first month by all assays, with a gradual fall in amount of IgM with time when assayed by μ-capture ELISA, a more gradual decline by IFA and hardly any decline with indirect ELISA. It was concluded that the indirect ELISA is unsuitable for the investigation of possible M. pneumoniae infection because the sustained high assay values with serum samples taken many months after infection, make interpretation of the test results very difficult.

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Galula, Jedhan Ucat, Gielenny M. Salem, Raul V. Destura, Roland Remenyi, and Day-Yu Chao. "Comparable Accuracies of Nonstructural Protein 1- and Envelope Protein-Based Enzyme-Linked Immunosorbent Assays in Detecting Anti-Dengue Immunoglobulin G Antibodies." Diagnostics 11, no. 5 (April 21, 2021): 741. http://dx.doi.org/10.3390/diagnostics11050741.

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Background: Dengue virus (DENV) infection remains a global public health concern. Enzyme-linked immunosorbent assays (ELISAs), which detect antibodies targeting the envelope (E) protein of DENV, serve as the front-line serological test for presumptive dengue diagnosis. Very few studies have determined the serostatus by detecting antibodies targeting the nonstructural protein 1 (NS1), which can function as diagnostic biomarkers to distinguish natural immunity from vaccine-induced immunity. Methods: We used community-acquired human serum specimens, with the serostatus confirmed by focus reduction microneutralization test (FRμNT), to evaluate the diagnostic performances of two NS1-based ELISA methods, namely, immunoglobulin G antibody-capture ELISA (NS1 GAC–ELISA) and indirect NS1 IgG ELISA, and compared the results with an E-based virus-like particle (VLP) GAC–ELISA. Results: NS1-based methods had comparable accuracies as VLP GAC–ELISA. Although the sensitivity in detecting anti-NS1 IgM was poor, indirect NS1 IgG ELISA showed similar limits of detection (~1–2 ng/mL) as NS1 GAC–ELISA in detecting anti-NS1 IgG. Combining the results from two or more tests as a composite reference standard can determine the DENV serostatus with a specificity reaching 100%. Conclusion: NS1-based ELISAs have comparable accuracies as VLP GAC–ELISA in determining dengue serostatus, which could effectively assist clinicians during assessments of vaccine eligibility.

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Kohl, Thomas O., and Carl A. Ascoli. "Indirect Competitive Enzyme-Linked Immunosorbent Assay (ELISA)." Cold Spring Harbor Protocols 2017, no. 7 (July 2017): pdb.prot093757. http://dx.doi.org/10.1101/pdb.prot093757.

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Veling, J., F. G. van Zijderveld, A. M. van Zijderveld-van Bemmel, H. W. Barkema, and Y. H. Schukken. "Evaluation of Three Newly Developed Enzyme-Linked Immunosorbent Assays and Two Agglutination Tests for Detecting Salmonella enterica subsp. enterica Serovar Dublin Infections in Dairy Cattle." Journal of Clinical Microbiology 38, no. 12 (2000): 4402–7. http://dx.doi.org/10.1128/jcm.38.12.4402-4407.2000.

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In this study test characteristics of three newly developed enzyme-linked immunosorbent assays (ELISAs) for Salmonella enterica subsp. enterica serovar Dublin were evaluated and compared with two agglutination tests. The ELISAs involved were an indirect ELISA with serovar Dublin lipopolysaccharide (LPS ELISA), an indirect ELISA with serovar Dublin flagellar antigen (GP ELISA), and a double-antibody sandwich blocking ELISA that uses monoclonal antibodies against S. enterica subsp.enterica serovar Enteritidis flagellin (GM-DAS ELISA). The agglutination tests involved were two routine serum agglutination tests with either somatic (O) or flagellar (H) antigen. Diagnostic specificity of the three ELISAs was determined using 840 serum samples from seven dairy herds without any history of serovar Dublin infection. Cutoff values at a titer of 100, 100, and 10, respectively, for the LPS ELISA, GP ELISA, and GM-DAS blocking ELISA resulted in a specificity of 99.3, 100, and 100%, respectively. Using these cutoff values the LPS ELISA, GP ELISA, and GM-DAS ELISA were able to detect, respectively, 30, 46, and 38% of 50 fecal culture-positive animals from 13 herds with a recent serovar Dublin infection. With the same cutoff values, active carriers (n = 18) were detected for 94.4% with the LPS ELISA and for 100% with the GP and GM-DAS ELISAs. Kappa values determined on the results of all tests from 8 of the 13 serovar Dublin-infected herds and the 7 control herds demonstrated a good correlation between the results of all ELISAs and the H-agglutination test. The results of the O-agglutination test failed to correlate with those of the other tests. Using a set of sera from 170 aborting cows (with 25 abortions due to serovar Dublin), test results of the ELISAs and the H-agglutination test were comparable. The H-agglutination test may be used successfully for single sample testing, especially to diagnose abortion due to serovar Dublin. It is concluded that the ELISAs are useful diagnostic tools in serovar Dublin control programs and that they are preferred to agglutination tests for reasons of automation and costs.

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Malan, Annette K., Erick Avelar, Sheldon E. Litwin, Harry R. Hill, and Christine M. Litwin. "Serological diagnosis of Trypanosoma cruzi: evaluation of three enzyme immunoassays and an indirect immunofluorescent assay." Journal of Medical Microbiology 55, no. 2 (February 1, 2006): 171–78. http://dx.doi.org/10.1099/jmm.0.46149-0.

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Chagas' disease is an important cause of heart failure in Latin America, but is rare in the United States. The immigration of persons from endemic countries increases the potential of encountering patients with the disease. Concerns have also been raised about the introduction of Trypanosoma cruzi, the parasite that causes the disease, into the blood supply and during organ transplantation. To compare Chagas' antibody tests that are available in the United States, we evaluated three IgG ELISAs, CeLLabs T. cruzi ELISA, Hemagen Chagas' kit and IVD Research Chagas' Serum Microwell ELISA, and MarDx indirect immunofluorescent assays. The CeLLabs and Hemagen IgG ELISAs had 100 % agreement, sensitivity and specificity. The IVD Research IgG ELISA had 94·6 % agreement, 100 % sensitivity and 93 % specificity.

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Van de gaer, Otto, Petra de Haes, and Xavier Bossuyt. "Detection of circulating anti-skin antibodies by indirect immunofluorescence and by ELISA: a comparative systematic review and meta-analysis." Clinical Chemistry and Laboratory Medicine (CCLM) 58, no. 10 (September 25, 2020): 1623–33. http://dx.doi.org/10.1515/cclm-2019-1031.

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AbstractBackgroundBoth enzyme-linked immunosorbent assays (ELISAs) and indirect immunofluorescence (IIF) are available for the diagnosis of autoimmune bullous diseases (AIBD). Many studies have reported on the performance of ELISAs and concluded that ELISAs could replace IIF. This study compares the diagnostic accuracy of ELISA and IIF for the detection of autoantibodies to desmoglein 1 (DSG1), desmoglein 3 (DSG3), bullous pemphigoid antigen 2 (BP180) and bullous pemphigoid antigen 1 (BP230) to support the diagnosis of pemphigus vulgaris (PV), pemphigus foliaceus (PF) and bullous pemphigoid (BP).MethodsA literature search was performed in the PubMed database. The meta-analysis was performed using summary values and a bivariate random effect model.ResultsThe five included studies on PV did not demonstrate significant differences between IIF and DSG3-ELISA (sensitivity 82.3% vs. 81.6%, p = 0.9284; specificity 95.6% vs. 93.9%, p = 0.5318; diagnostic odds ratio [DOR] 101.60 vs. 67.760, p = 0.6206). The three included studies on PF did not demonstrate significant differences between IIF and DSG1-ELISA (sensitivity 80.6% vs. 83.1%, p = 0.8501; specificity 97.5% vs. 93.9%, p = 0.3614; DOR 160.72 vs. 75.615, p = 0.5381). The eight included studies on BP showed that BP230-ELISA differed significantly from both IIF on monkey esophagus (MO) and BP180-ELISA with regard to DOR (11.384 vs. 68.349, p = 0.0008; 11.384 vs. 41.699, p = 0.0125, respectively)ConclusionsOur meta-analysis shows that ELISA performs as well as IIF for diagnosing PV, PF and BP.

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Rhodes, Marvin B., Carol A. Klucas, Merwin L. Frey, and Gary A. Anderson. "A Blocking ELISA for the Detection of Specific Antibodies to Bovine Respiratory Syncytial Virus." Journal of Veterinary Diagnostic Investigation 1, no. 4 (October 1989): 324–28. http://dx.doi.org/10.1177/104063878900100408.

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A blocking enzyme-linked immunosorbent assay (ELISA) has been adapted to detect specific antibodies in bovine sera to respiratory syncytial virus using a horseradish peroxidase-labeled monclonal antibody to the fusion protein of the virus. This assay plus an indirect blocking ELISA and indirect ELISA were used to detect antibodies to the bovine respiratory syncytial virus (BRSV) in 159 field-origin bovine sera. Results of these assays were compared with serum antibody titers measured by the serum neutralization (SN) test. Over a 56-day period, the mean neutralization titers and the mean delta absorbance values for the blocking ELISA, on the same sera, showed similar declines. However, the calculated correlation coefficients between mean SN titer and mean absorbance value for the blocking ELISA of the individual sera ranged from −0.2 to −0.5 depending on the source of sera. Similar values were obtained whether using crude or purified viral antigen in the assays. Corresponding calculated correlation coefficients were generally higher for the indirect blocking ELISA or indirect ELISA than for the blocking ELISA. The blocking ELISA was between 70 and 64% as sensitive as the serum neutralization test with a specificity of 100 or 90% using the crude and purified viral antigen, respectively. The indirect blocking ELISA and indirect ELISA had similar calculated sensitivities and specificities. The blocking ELISA was faster to run than either of the other ELISA's or the neutralization test. Further, nonspecific background absorbance was obviated because the blocking ELISA detects antibodies to 1 specific viral protein, the fusion protein. These studies suggest that the blocking ELISA should be useful as a serological test for BRSV antibodies.

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Petraitytė, Rasa, Li Jin, Rashpal Hunjan, Aušra Ražanskienė, Aurelija Žvirblienė, and Kęstutis Sasnauskas. "Use of Saccharomyces cerevisiae-Expressed Recombinant Nucleocapsid Protein To Detect Hantaan Virus-Specific Immunoglobulin G (IgG) and IgM in Oral Fluid." Clinical and Vaccine Immunology 14, no. 12 (October 3, 2007): 1603–8. http://dx.doi.org/10.1128/cvi.00188-07.

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ABSTRACT Hantaan virus is the causative agent of severe hemorrhagic fever with renal syndrome. Clinical surveillance for Hantaan virus infection is unreliable, and laboratory verification is essential. The detection of virus-specific immunoglobulin M (IgM) and IgG in serum is most commonly used for the diagnosis of hantavirus infection. Testing of oral fluid samples instead of serum offers many advantages for surveillance. However, commercial tests for hantavirus-specific antibodies are unavailable. For the detection of Hantaan virus in the oral fluid of humans, we have developed a monoclonal antibody-based capture enzyme-linked immunosorbent IgM assay (IgM capture ELISA) and indirect enzyme-linked immunosorbent IgG and IgM assays (indirect IgG and IgM ELISAs) for paired serum and oral fluid samples using the Saccharomyces cerevisiae yeast-expressed nucleocapsid protein of the Hantaan-Fojnica virus. The sensitivity and specificity of the oral fluid IgM capture ELISA in comparison with the results of the serum Hantaan virus IgM assay were 96.7% and of 94.9%, respectively. Thus, data on the overall performance of the oral fluid IgM capture ELISA are in close agreement with those of the serum IgM assay, and the method exhibits the potential to serve as an easily transferable tool for large-scale epidemiological studies. Data on the indirect IgM ELISA also showed close agreement with the serum IgM assay data; however, the indirect IgG ELISA displayed a lower sensitivity and a lower specificity. In conclusion, the IgM capture ELISA can be used with oral fluid instead of serum samples for the diagnosis of Hantaan virus infection.

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Wilson, Albert J., Howard Sant, Peter K. Van Duser, and Myron Wentz. "Enzyme-Based Methods for IgM Serology: Standard Indirect ELISA vs Antibody-Capture ELISA." Laboratory Medicine 23, no. 4 (April 1, 1992): 259–63. http://dx.doi.org/10.1093/labmed/23.4.259.

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Suh, Lindsey Y. K., Tayabaa Kartoon, Naiyana Gujral, Youngmee Yoon, Joo Won Suh, and Hoon Sunwoo. "The Use of Chicken Igy in a Double Antibody Sandwich Elisa for the Quantification of Melittin in Bee Venom and Bee Venom Melittin Content in Cosmetics." Journal of Apicultural Science 59, no. 1 (June 1, 2015): 97–107. http://dx.doi.org/10.1515/jas-2015-0011.

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Abstract Two enzyme-linked immunosorbent assay (ELISA) - based detection systems: indirect competitive ELISA and biotinylated double antibody sandwich ELISA (DAS-ELISA) were developed to determine the melittin concentration in honeybee (Apis mellifera) venom and the melittin concentration in cosmetics which contain bee venom. The indirect competitive ELISA employed chicken anti-melittin IgY. The biotinylated DAS-ELISA employed anti-melittin monoclonal antibody (MAb) and biotinylated anti-melittin IgY. To produce anti-melittin IgY; Sigma melittin was emulsified with Freund‘s incomplete adjuvant and immunised to Leghorn laying chickens intramuscularly at four different sites (50 μg/mL, 0.25 mL per site) of the breast muscles. After 5 to 8 weeks of the immunisation, anti-melittin IgY was extracted and analysed by ELISA. The anti-melittin IgY antibody produced was highly specific to melittin and did not cross-react with other bee venom proteins, as examined by ELISA and a western-blot assay. Indirect competitive ELISA demonstrated a higher range of melittin detection (2.5 to 80 μg/mL). Double antibody sandwich ELISA using MAb as the capture antibody and biotinylated polyclonal IgY as the detection antibody, provided a lower range of detection (2.5 - 40 ng/mL), which has a 1000 times higher sensitivity than that of indirect competitive ELISA. Therefore, indirect competitive ELISA is a useful tool to measure the concentration of melittin in bee venom as a raw material. Biotinylated DAS-ELISA, on the other hand, is more suitable for nanoscale quantification of melittin in commercial products.

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NUNES, Cáris Maroni, Regina Nardini TUNDISI, Marcos Bryan HEINEMANN, Saemi OGASSAWARA, and Leonardo José RICHTZENHAIN. "TOXOCARIASIS: SEROLOGICAL DIAGNOSIS BY INDIRECT ANTIBODY COMPETITION ELISA." Revista do Instituto de Medicina Tropical de São Paulo 41, no. 2 (March 1999): 95–100. http://dx.doi.org/10.1590/s0036-46651999000200006.

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Toxocariasis is caused by infection of man by Toxocara canis and Toxocara cati larvae, the common roundworm of dogs and cats. Because larvae are difficult to detect in tissues, diagnosis is mostly based on serology. Non specific reactions are observed mainly due to cross-reactivity with Ascaris sp antigens. This investigation aimed at developing and evaluating an indirect antibody competition ELISA (IACE) employing a specific rabbit IgG anti-Toxocara canis excretory-secretory antigens as the competition antibody, in order to improve indirect ELISA specificity performed for toxocariasis diagnosis. For that, the rabbit IgG was previously absorbed by Ascaris suum adult antigens. Sensitivity and specificity of IACE were first evaluated in 28 serum samples of mice experimentally infected with T. canis embryonated eggs. Adopting cut-off value established in this population before infection, sensitivity and specificity were 100% after 20 days post-inoculation. For human population IACE was evaluated using sera from 440 patients with clinical signs of toxocariasis and the cut-off value was established with 60 serum samples from apparently healthy individuals. Using as reference test the indirect ELISA performed by Adolfo Lutz Institute, sensitivity was 60.2%, specificity was 98% and concordance was 77.3%. Repeatability of IACE was evaluated by the inter-reactions variation coefficient (2.4%).

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LI, YONGNING, YUANYUAN WANG, and YANGHAO GUO. "AN INDIRECT COMPETITIVE ELISA FOR DETERMINATION OF CITRININ." Journal of Food Safety 31, no. 4 (October 24, 2011): 497–504. http://dx.doi.org/10.1111/j.1745-4565.2011.00326.x.

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HOCHEL, IGOR, and MAREK MUSIL. "Development of an Indirect Competitive ELISA of DDT." Food and Agricultural Immunology 14, no. 4 (December 2002): 285–300. http://dx.doi.org/10.1080/0954010021000096391.

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Afshar, A., G. C. Dulac, P. F. Wright, and D. Martin. "Application of Indirect ELISA for Detection of Bovine Antibodies against Vesicular Stomatitis Viruses." Journal of Veterinary Diagnostic Investigation 5, no. 1 (January 1993): 26–32. http://dx.doi.org/10.1177/104063879300500107.

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Two indirect enzyme-linked immunosorbent assays (I-ELISAs) are described for the detection of bovine serum antibody to the New Jersey (NJ) and Indiana (IN) vesicular stomatitis viruses (VSV). Serum samples at a dilution of 1:200 were incubated with binary ethylenimine-inactivated VSV-NJ and VSV-IN type-specific antigens preadsorbed to microtiter plates. Bound antibodies were detected by a murine monoclonal antibody to bovine IgG 1 conjugated with horseradish peroxidase. The performance of each I-ELISA in detecting homotypic and heterotypic antibodies to VSV-NJ and VSV-IN in sequential serum samples from calves experimentally infected with VSV-NJ or VSV-IN was evaluated. The I-ELISAs detected serotype-specific antibodies to either VSV-NJ or VSV-IN in calves infected with the homologous serotype. Homotypic but not heterotypic anti-VSV-NJ antibodies were first demonstrable by the VSV-NJ I-ELISA during the second week postinfection and remained at an elevated level for a period of 11 weeks, with a gradual decrease thereafter. Similar homotypic antibody profiles measured by the VSV-IN I-ELISA in calves inoculated with VSV-IN were observed. The performances of the I-ELISAs were compared using 1,495 microtiter serum neutralization (MTSN) test-negative bovine field sera collected from cattle in Canada (VS free) and 429 samples collected from cattle in the USA and Mexico (VS-epidemic and VS-endemic areas). The diagnostic specificities of the VSV-NJ and VSV-IN I-ELISAs for the Canadian samples relative to the MTSN test results were in the range of 99.8% and 99.7%, respectively. The levels of agreement between the VSV-NJ and VSV-IN I-ELISAs and the MTSN test for the 429 bovine field samples from VS-epidemic and VS-endemic areas were 97.7% and 98.8%, respectively. Relative to MTSN, the sensitivity and specificity of the 429 field sera for the VSV-NJ I-ELISA were 95.4% and 99.6%, respectively, and for the VSV-IN assay were 75.0% and 99.2%, respectively. The results suggest that in addition to the many technical advantages over the MTSN test these I-ELISAs have potential application as rapid and inexpensive tests for the serodiagnosis of VSV infection in cattle.

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M.Sasikala, K.R.Chithra, J.J.Solomon, and G.Rajeev. "DEVELOPMENT OF DAC INDIRECT ELISA FOR THE RAPID DETECTION OF COCONUT ROOT (WILT) DISEASE." CORD 17, no. 02 (June 1, 2001): 34. http://dx.doi.org/10.37833/cord.v17i02.350.

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Identification of root(wilt) disease-free palm is a basic requirement for evolving disease resistant/tolerant planting materials for the management of this phytoplasma induced malady. For this purpose, various types of ELISA were performed with different enzymes and their respective substrates to standardize the most suitable and sensitive one. Indirect ELISA, Protein A indirect ELISA and F(ab')2 indirect ELISA using Alkaline Phosphatase conjugate and DAC indirect ELISA using Horse raddish peroxidase conjugate were carried out. Of the various types of ELISA tried, DAC indirect ELISA has been found to be the best for the rapid detection of coconut root(wilt) disease. In this assay, crude leaf extracts and unfractionated polyclonal antiserum were employed as test antigen and specific antibody respectively. Higher specificity was observed with the addition of gelatin and ovalbumin in the extraction medium and overnight incubation of ELISA plate at 5°C after substrate addition. Antigen titre was found to be very high in spear leaves followed by the next outer leaf. Similarly, maximum antigen titre was observed during the early stage of the diseased palm. The test could be completed within 44h and in a single ELISA plate, 20 samples with three replications could be screened using microlitre quantities of the specific antibody.

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Attia, Marwa M., Elshaimaa Ismael, and Nagla M. K. Saleh. "A sensitive serodiagnostic tool for the detection of active infection of zoonotic visceral and nasopharyngeal linguatulosis." Veterinary World 12, no. 6 (June 2019): 883–89. http://dx.doi.org/10.14202/vetworld.2019.883-889.

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Aim: This study aimed to evaluate the different serological techniques for early diagnosis of acute concurrent infections with linguatulosis in the definitive host (dogs) and an intermediate host (goats). This evaluation compared between the gold standard (GS) test (GS; examination of nasal and fecal samples in dogs and examination of lymph nodes in goats), sandwich enzyme-linked immunosorbent assay (S-ELISA), and indirect ELISA. Materials and Methods: Fifty goats and fifty dogs were examined for the presence of Linguatula serrata nymphs and adults, respectively, besides the collection of blood samples from the examined animals for serologic testing. Results: In goats; GS, S-ELISA, and indirect ELISA showed positivity in 32 (64%), 28 (56%), and 39 (78%) samples, respectively. In dogs; GS, S-ELISA, and indirect ELISA showed positivity in 25 (50%), 25 (50%), and 30 (60%) samples, respectively. S-ELISA displayed significant higher agreement with the GS test (≥0.83) than indirect ELISA (≤0.67) in both hosts. Infection with linguatulosis showed significant relation with the age of goats and dogs and the sex of goats (p<0.05). Conclusion: S-ELISA displayed more sensitivity and specificity for the detection of concurrent infections with linguatulosis in both dogs and goats than indirect ELISA, which could detect the prior infections. Similarly, these assays could be used for diagnosis of concurrent infections with linguatulosis in human, especially the chronic ones.

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Martínez-Torrecuadrada, Jorge L., Beatriz Lázaro, José F. Rodriguez, and J. Ignacio Casal. "Antigenic Properties and Diagnostic Potential of Baculovirus-Expressed Infectious Bursal Disease Virus Proteins VPX and VP3." Clinical Diagnostic Laboratory Immunology 7, no. 4 (July 1, 2000): 645–51. http://dx.doi.org/10.1128/cdli.7.4.645-651.2000.

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ABSTRACT The routine technique for detecting antibodies specific to infectious bursal disease virus (IBDV) is a serological evaluation by enzyme-linked immunosorbent assay (ELISA) with preparations of whole virions as the antigens. To avoid using complete virus in the standard technique, we have developed two new antigens through the expression of the VPX and VP3 genes in insect cells. VPX and especially VP3 were expressed at high levels in insect cells and simple to purify. The immunogenicity of both proteins was similar to that of the native virus. VPX was able to elicit neutralizing antibodies but VP3 was not. Purified VPX and VP3 were tested in an indirect ELISA with more than 300 chicken sera. There was an excellent correlation between the results of the ELISA using VPX and those of the two commercial kits. VP3 did not perform as well as VPX, and the linear correlation was significantly lower. A comparison with the standard reference technique, seroneutralization, showed that the indirect ELISA was more sensitive. Therefore, VPX-based ELISA is a good alternative to conventional ELISAs that use whole virions.

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Xin, Ting, Hongjun Yang, Nan Wang, Fang Wang, Peng Zhao, Haiguang Wang, Kairong Mao, Hongfei Zhu, and Jiabo Ding. "Limitations of the BP26 Protein-Based Indirect Enzyme-Linked Immunosorbent Assay for Diagnosis of Brucellosis." Clinical and Vaccine Immunology 20, no. 9 (July 17, 2013): 1410–17. http://dx.doi.org/10.1128/cvi.00052-13.

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ABSTRACTBrucellosis is a serious zoonosis that occurs worldwide, and its diagnosis is typically based on the detection of antibodies againstBrucellalipopolysaccharide (LPS). However, the specificity of the LPS-based test is compromised by cross-reactivity withEscherichia coliO157:H7 andYersinia enterocoliticaO:9. Also, diagnosis based on the LPS test cannot differentiate between vaccinated and infected individuals. The detection of the 26-kDa cytosoluble protein (BP26) antibody is considered an alternative that circumvents these drawbacks because it is exclusively expressed by infectiousBrucella. A BP26-based enzyme-linked immunosorbent assay (ELISA) has been tried for the diagnosis ofBrucella-infected animals and humans, but a few results showed that BP26 couldn't react with allBrucella-positive sera. In order to explore whether different animals could produce antibodies against BP26 after being infected with variousBrucellaspecies, we infected sheep, goats, and beef cattle with common virulent referenceBrucellaspecies. All sera were collected from the experimental animals and tested using both LPS-based ELISAs and BP26-based ELISAs. The results showed that allBrucella-infected individuals could produce high levels of antibodies against LPS, but onlyB. melitensis16M- andB. melitensisM28-infected sheep andB. melitensis16M- andB. abortus2308-infected goats could produce antibodies against BP26. Therefore, we concluded that the BP26-based indirect ELISA (i-ELISA) showed bothBrucellaspecies and host specificity, which obviously limits its reliability as a substitute for the traditional LPS-based ELISA for the detection of brucellosis.

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Xie, Songhua, Qiuping Shen, Wei Zhang, Weikang Wang, Quan Xie, Tuofan Li, Zhimin Wan, Hongxia Shao, Aijian Qin, and Jianqiang Ye. "An efficient peptide-based ELISA for differentiating fowl adenovirus 4–infected chickens from vaccinated chickens." Journal of Veterinary Diagnostic Investigation 33, no. 4 (April 15, 2021): 762–66. http://dx.doi.org/10.1177/10406387211005749.

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Fowl adenovirus serotype 4 (FAdV4), the causative agent of hepatitis-hydropericardium syndrome (HPS), has caused major economic losses to the poultry industry worldwide. Although inactivated vaccines have been deployed widely against FAdV4, a DIVA (differentiating infected from vaccinated animals) test specific for FAdV4 has not been available. We synthesized an immunogenic peptide, corresponding to regions 66–88 aa of the 22K nonstructural protein of FAdV4, and used the peptide as coating antigen to develop an indirect ELISA for a DIVA test specific to FAdV4. Specificity analysis showed that the ELISA only reacted with sera against FAdV4, and not with sera against other pathogens tested. Moreover, the ELISA could effectively differentiate FAdV4–infected chickens from vaccinated chickens. In a test of sera from experimentally infected chickens, the ELISA had 95% and 85% concordance with an indirect immunofluorescence assay (indirect IFA) and a commercial ELISA, respectively, and the concordance was 80.5% between the ELISA and the indirect IFA in detecting clinical infection samples. Our peptide-based ELISA provides an efficient DIVA test for FAdV4 in clinical samples.

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Tenter, A. M., C. Vietmeyer, A. M. Johnson, K. Janitschke, M. Rommel, and W. Lehmacher. "ELISAs based on recombinant antigens for sero-epidemiological studies onToxoplasma gondiiinfections in cats." Parasitology 109, no. 1 (July 1994): 29–36. http://dx.doi.org/10.1017/s0031182000077738.

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SUMMARYTwo recombinantToxoplasma gondiipolypeptides, H4 and H11, were tested as diagnostic antigens in enzyme-linked immunosorbent assays (ELISAs). The results obtained by ELISAs based on single H4 (H4-ELISA), on single H11 (H11-ELISA) and on a mixture of H4 and H11 (H4/H11-ELISA) were compared with results obtained by an ELISA based on traditional ELISA antigen (TEA-ELISA), an indirect fluorescent antibody test (IFAT), the Sabin-Feldman dye test (SFDT) and a direct agglutination test (DAT). A total of 306 cats from a suburban cat population were tested of which about 45% showed serological evidence ofT. gondiiinfection. Infection rates varied from about 32% for cats kept indoors to about 55% for stray cats. Specificities > 99% were observed for all ELISAs based on the recombinant antigens (H4-ELISA, H11-ELISA and H4/H11-ELISA). The H4/H11-ELISA also reached a sensitivity of 95% which compared very favourably with those observed for the TEA-ELISA (98%) and for the IFAT (94%). Negative and positive predictive values for the H4/H11-ELISA were 96 and < 99%, respectively. Antibody titres measured by the H4/H11-ELISA also correlated well with those measured by the SFDT and the DAT. Hence, the H4/H11-ELISA appears to be a very suitable test for sero-epidemiological studies onT. gondiiinfections in cats.

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Carvalho, Fernando R., Deise A. O. Silva, Jair P. Cunha-Júnior, Maria A. Souza, Taísa C. Oliveira, Samantha R. Béla, Gabriele G. Faria, Carolina S. Lopes, and José R. Mineo. "Reverse Enzyme-Linked Immunosorbent Assay Using Monoclonal Antibodies against SAG1-Related Sequence, SAG2A, and p97 Antigens from Toxoplasma gondii To Detect Specific Immunoglobulin G (IgG), IgM, and IgA Antibodies in Human Sera." Clinical and Vaccine Immunology 15, no. 8 (June 18, 2008): 1265–71. http://dx.doi.org/10.1128/cvi.00069-08.

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ABSTRACT The present study aimed to evaluate the performance of three monoclonal antibodies (MAbs) in reverse enzyme-linked immunosorbent assays (ELISAs) for detecting immunoglobulin G (IgG), IgM, and IgA antibodies against Toxoplasma gondii in 175 serum samples from patients at different stages of T. gondii infection, as defined by both serological and clinical criteria, as follows: recent (n = 45), transient (n = 40), and chronic (n = 55) infection as well as seronegative subjects (n = 35). The results were compared with those obtained by indirect ELISA using soluble Toxoplasma total antigen (STAg). Our data demonstrated that MAb A3A4 recognizes a conformational epitope in SAG1-related-sequence (SRS) antigens, while A4D12 and 1B8 recognize linear epitopes defined as SAG2A surface antigen and p97 cytoplasmatic antigen, respectively. Reverse ELISA for IgG with A3A4 or A4D12 MAbs was highly correlated with indirect ELISA for anti-STAg IgG, whereas only A4D12 reverse ELISA showed high correlation with indirect ELISA for IgM and IgA isotypes. To our knowledge, this is the first report analyzing the performance of a reverse ELISA for simultaneous detection of IgG, IgM, and IgA isotypes active toward native SAG2A, SRS, and p97 molecules from STAg, using a panel of human sera from patients with recent and chronic toxoplasmosis. Thus, reverse ELISA based on the capture of native SAG2A and SRS antigens of STAg by MAbs could be an additional approach for strengthening the helpfulness of serological tests assessing the stage of infection, particularly in combination with highly sensitive and specific assays that are frequently used nowadays for diagnosis of toxoplasmosis during pregnancy or congenital infection in newborns.

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Ghurafa, Rinaldi, Denny Widaya Lukman, and Hadri Latif. "Indirect Enzyme Linked Immunosorbent Assay Sebagai Metode untuk Melacak Bruselosis pada Sapi Perah (INDIRECT ENZYME IMMUNOSORBENT ASSAY (I-ELISA) AS METHOD FOR DETECT BRUCELLOSIS IN DAIRY COW)." Jurnal Veteriner 20, no. 1 (May 24, 2019): 30. http://dx.doi.org/10.19087/jveteriner.2019.20.1.30.

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Brucellosis has become a zoonotic disease that received attention in efforts to prevent and eradicate strategic infectious animal diseases in Indonesia. Brucellosis can be detected early by the rose bengal test (RBT), followed by complement fixation test (CFT) and by enzyme linked immunosorbent assay (ELISA). The aims of this research was to study the indirect enzyme linked immunosorbent assay test (I-ELISA) as an alternative test for detecting brucellosis in dairy cattle. The method was used by conducting tests of RBT, CFT, I-ELISA and commercial I-ELISA to test brucellosis. The test results were calculated sensitivity and specificity, as well as analyzed by calculating the kappa value. The method was used by conducting tests of RBT, CFT, I-ELISA and commercial I-ELISA to test brucellosis. The test results were calculated for sensitivity and specificity, as well as analyzed by calculating the Kappa statistical value. The results of the sensitivity and specificity calculation showed that the indirect enzyme linked immunosorbent assay (I-ELISA) test developed a higher sensitivity (100%) compared to RBT test (93.75%) and commercial I-ELISA (93.75%). The developed I-ELISA specificity (74.68%) was still lower than RBT (89.87%), but higher than commercial I-ELISA (70.52%). The calculation of the statistical value of kappa RBT with CFT showed the kappa value 0.7120 which meaned it had a good agreement, commercial I-ELISA with CFT showed kappa value 0.6165 which meaned it had good suitability, whereas I-ELISA developed with CFT showed kappa value 0.4984 which meaned having a moderate agreement.In conclusion, the indirect enzyme linked immunosorbent assay (I-ELISA) which had been developed had low specificity, but the sensitivity was the highest compared to the commercial I-ELISA test and RBT, so this test was appropriate to be used as a screening test, especially in dairy cows movement into brucellosis-free areas or regions.

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Emmerich, Petra, Ronald von Possel, Christina Deschermeier, Salih Ahmeti, Lindita Berisha, Bahrije Halili, Xhevat Jakupi, Kurtesh Sherifi, Claudia Messing, and Viola Borchardt-Lohölter. "Comparison of diagnostic performances of ten different immunoassays detecting anti-CCHFV IgM and IgG antibodies from acute to subsided phases of Crimean-Congo hemorrhagic fever." PLOS Neglected Tropical Diseases 15, no. 3 (March 15, 2021): e0009280. http://dx.doi.org/10.1371/journal.pntd.0009280.

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Crimean-Congo Hemorrhagic Fever Virus (CCHFV) is a geographically widespread tick-borne arbovirus that has been recognized by the WHO as an emerging pathogen needing urgent attention to ensure preparedness for potential outbreaks. Therefore, availability of accurate diagnostic tools for identification of acute cases is necessary. A panel comprising 121 sequential serum samples collected during acute, convalescent and subsided phase of PCR-proven CCHFV infection from 16 Kosovar patients was used to assess sensitivity. Serum samples from 60 healthy Kosovar blood donors were used to assess specificity. All samples were tested with two IgM/IgG immunofluorescence assays (IFA) from BNITM, the CCHFV Mosaic 2 IgG and IgM indirect immunofluorescence tests (IIFT) from EUROIMMUN, two BlackBox ELISAs for the detection of CCHFV-specific IgM and IgG antibodies (BNITM), two Anti-CCHFV ELISAs IgM and IgG from EUROIMMUN using recombinant structural proteins of CCHFV antigens, and two ELISAs from Vector-Best (IgM: μ-capture ELISA, IgG: indirect ELISA using immobilized CCHFV antigen). Diagnostic performances were compared between methods using sensitivity, specificity, concordance and degree of agreement with particular focus on the phase of the infection. In early and convalescent phases of infection, the sensitivities for detecting specific IgG antibodies differed for the ELISA test. The BlackBox IgG ELISA yielded the highest, followed by the EUROIMMUN IgG ELISA and finally the VectorBest IgG ELISA with the lowest sensitivities. In the subsided phase, the VectorBest IgM ELISA detected a high rate of samples that were positive for anti-CCHFV IgM antibodies. Both test systems based on immunofluorescence showed an identical sensitivity for detection of anti-CCHFV IgM antibodies in acute and convalescent phases of infection. Available serological test systems detect anti-CCHFV IgM and IgG antibodies accurately, but their diagnostic performances vary with respect to the phase of the infection.

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Toaleb, Nagwa I., Mohamed S. Helmy, Eman E. El Shanawany, and Eman H. Abdel-Rahman. "A simple and efficient purification method of native immunoreactive antigen for diagnosis of camel hydatidosis." January-2020 13, no. 1 (2020): 141–46. http://dx.doi.org/10.14202/vetworld.2020.141-146.

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Background: Cystic echinococcosis (CE), a zoonotic disease that affects animal and human health, is of increasing economic importance due to high morbidity rates and high economic losses in the livestock industry. Aim: The present study was conducted to purify the antigen from hydatid cyst fluid (HCF) with high diagnostic efficacy of camel hydatidosis using indirect enzyme-linked immunosorbent assay (ELISA). Materials and Methods: The HCF antigen was purified using Sephacryl S-300 column chromatography. Characterization of fractions was performed using reducing and non-reducing sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis. Further, antibodies against Echinococcus granulosus cysts in camel serum were detected using indirect ELISA. Results: The purification process resulted in three fractions of antigens: FI, FII, and FIII. Indirect ELISA showed that higher diagnostic efficacy was observed in FI than in FII and FIII. Indirect ELISA, in which FI was utilized, showed 88% sensitivity and 91.7% specificity. Non-reducing SDS-PAGE showed that FI had two bands of molecular weights 120 and 60 kDa. Western blot analysis of FI demonstrated that 60, 38, and 22 kDa were antigenic bands when reacted with naturally infected camel sera with E. granulosus cysts. Using indirect ELISA, F1 recorded an infection percentage of 81.7% in randomly collected camel serum samples. Conclusion: FI is a promising antigen for accurate diagnosis of camel CE using indirect ELISA.

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Li, Hong, Travis C. McGuire, Uwe U. Müller-Doblies, and Timothy B. Crawford. "A Simpler, More Sensitive Competitive Inhibition Enzyme-Linked Immunosorbent Assay for Detection of Antibody to Malignant Catarrhal Fever Viruses." Journal of Veterinary Diagnostic Investigation 13, no. 4 (July 2001): 361–64. http://dx.doi.org/10.1177/104063870101300417.

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An earlier competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA) was developed for detection of specific antibody against malignant catarrhal fever (MCF) viruses (MCFV) in ruminants. In this study, the indirect CI-ELISA was improved by conjugating the monoclonal antibody 15-A directly with horseradish peroxidase and by developing a method of producing precoated, dried antigen plates. This new test is referred to as a direct CI-ELISA. The reformatted test yielded a significantly improved sensitivity, and the time required was reduced to about one-sixth of the previous time. Of 37 MCF cases in cattle that were confirmed by histopathology and polymerase chain reaction (PCR) assay, 37 (100%) were positive by the new test, whereas the indirect CI-ELISA detected only 23 (62%). The direct CI-ELISA detected antibody to MCFV in 100% of 48 sheep that had been defined as infected with ovine herpesvirus 2 (OvHV-2) by PCR, whereas the indirect CI-ELISA detected only 41 (85%). Comparison of antibody titers measured by the 2 assays for sera collected from OvHV-2-infected sheep and from cattle, bison, and deer with clinical sheep-associated MCF revealed that the direct CI-ELISA offered a 4-fold increase in analytical sensitivity over the indirect format. The number of seropositive animals detected by the direct CI-ELISA among apparently normal cattle and bison was 2–3 times greater than the number detected by the indirect CI-ELISA, indicating that a significant percentage of normal cattle and bison are subclincally infected with MCFV.

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A. Husain, Mahmood, Mahmood M. Nasser, and Mohmad M. Abiad. "Diagnosis of Autoimmune Hepatitis By ELISA and Indirect IF." Rafidain Journal of Science 21, no. 1 (January 28, 2010): 62. http://dx.doi.org/10.33899/rjs.2010.38775.

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Liu, Yue Hui, Jun Wang, Xin Chao Yang, Na Xin Sun, and Li Hua Xu. "Performance of Indirect ELISA for Maize Hybrid Purity Test." Advanced Materials Research 781-784 (September 2013): 1741–44. http://dx.doi.org/10.4028/www.scientific.net/amr.781-784.1741.

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Maize is the second large crop in China that needed 100-120 million tons seeds for an average year and the production loss caused by inferior seeds was amazing. Bing an allogamous wind-pollinated plant, maize hybrid purity problem is mainly reflected the inbred lines seed mixed in hybrid. Therefore it is important to have specific and sensitive purity test methods to prevent the poor seeds into market. In this paper, we developed an inderect ELISA using polyconal antibody against maize inbred lines to bind extracts from whole grain with pure water. The single blind trial demonstrated that the average detectable rate of indrect ELISA was 95.2%. We also extended this method to other maize hybrid varieties, found it has no ability to descriminate the maize hybrid facticity.

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M. Ahmed, I. "Serodiagnosis of Johne's disease by indirect ELISA in ovine." Iraqi Journal of Veterinary Sciences 24, no. 1 (June 28, 2010): 41–43. http://dx.doi.org/10.33899/ijvs.2010.5576.

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Krishnasamy, Kaveri, Rohan Narayan, Senthil Raja, Senthil Kumar, Mohana Sambasivam, Raja Jagadeesan, Kavita Arunagiri, and Gunasekaran Palani. "A novel indirect ELISA for diagnosis of dengue fever." Indian Journal of Medical Research 144, no. 1 (2016): 128. http://dx.doi.org/10.4103/0971-5916.193300.

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Kolhe, R. P., K. N. Bhilegaonkar, Z. B. Dubbal, Simranpreet Kaur, Samir Das, and R. K. Agarwal. "Indirect ELISA for serosurveillance of Japanese encephalitis in pigs." Indian Journal of Animal Research 49, no. 3 (2015): 343. http://dx.doi.org/10.5958/0976-0555.2015.00065.5.

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Pelonero, AnthonyL, AnandK Pandurangi, and VincentP Calabrese. "Viral antibodies in schizophrenia using the indirect elisa method." Schizophrenia Research 2, no. 1-2 (January 1989): 171. http://dx.doi.org/10.1016/0920-9964(89)90207-7.

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Chan, Lawrence S. "ELISA Instead of Indirect IF in Patients With BP." Archives of Dermatology 147, no. 3 (March 1, 2011): 291. http://dx.doi.org/10.1001/archdermatol.2011.24.

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O'Loan, Colette J., G. M. Allan, J. McNair, D. P. Mackie, and M. S. McNulty. "TRT virus serology: Discrepancy between ELISA and indirect immunofluorescence." Avian Pathology 19, no. 1 (January 1990): 173–80. http://dx.doi.org/10.1080/03079459008418667.

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Sivajothi, S., V. C. Rayulu, P. Malakondaiah, and D. Sreenivasulu. "Diagnosis of Trypanosoma evansi in bovines by indirect ELISA." Journal of Parasitic Diseases 40, no. 1 (April 26, 2014): 141–44. http://dx.doi.org/10.1007/s12639-014-0465-z.

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Marín, C. M., E. Moreno, I. Moriyón, R. Díaz, and J. M. Blasco. "Performance of Competitive and Indirect Enzyme-Linked Immunosorbent Assays, Gel Immunoprecipitation with Native Hapten Polysaccharide, and Standard Serological Tests in Diagnosis of Sheep Brucellosis." Clinical Diagnostic Laboratory Immunology 6, no. 2 (March 1, 1999): 269–72. http://dx.doi.org/10.1128/cdli.6.2.269-272.1999.

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ABSTRACT Competitive and standard enzyme-linked immunosorbent assays (ELISAs), rose bengal (RB), complement fixation, and agar gel immunoprecipitation with native hapten (AGID-NH) were compared by using sera from Brucella-free, Brucella melitensis-infected, and B. melitensisRev1-vaccinated sheep. The most sensitive tests were indirect ELISA and RB, and the most specific tests were AGID-NH and competitive ELISA. We show that RB followed by AGID-NH is a simple and effective system for diagnosing sheep brucellosis.

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Elderbrook, Molly J., Brant A. Schumaker, Massaro W. Ueti, Meila Bastos de Almeida, Thallitha S. W. J. Vieira, Rafael F. C. Vieira, and Kerry S. Sondgeroth. "Comparison of 2 ELISAs for detecting exposure to Brucella ovis." Journal of Veterinary Diagnostic Investigation 32, no. 5 (August 4, 2020): 700–705. http://dx.doi.org/10.1177/1040638720943880.

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Control of Brucella ovis infection in sheep flocks in the United States depends on early detection of B. ovis antibodies via serologic testing. We used 2,276 sheep sera and various cutoff values to compare seroprevalence and agreement between 2 ELISAs: the National Veterinary Services Laboratories (NVSL) B. ovis indirect ELISA and the IDEXX B. ovis ELISA kit. A subset of 295 sera was used to compare agreement and evaluate relative sensitivity and specificity of the 2 ELISAs with an agar gel immunodiffusion (AGID) test kit. There was no significant difference in B. ovis seroprevalence between the ELISAs; however, there was poor agreement between them. When the AGID test was used as the reference test, the IDEXX ELISA with a moderate cutoff value (S/P ratio = 45%) had the highest relative sensitivity of 38.1% and specificity of 92.0%. The NVSL ELISA with a lax cutoff value (S/P ratio = 0.75) had relative sensitivity of 19.1% and specificity of 94.6%. Receiver operating characteristic analysis revealed that optimal cutoff values for the NVSL and IDEXX ELISAs were 0.091 and 16.5%, respectively. This results in sensitivity and specificity of 85.7% and 31.8% for the NVSL ELISA, and sensitivity and specificity of 81.0% and 53.6% for the IDEXX ELISA, respectively.

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XU, Yi-Chun, Guang S. Zhang, and Fun S. Chu. "Enzyme-Linked Immunosorbent Assay for Deoxynivalenol in Corn and Wheat." Journal of AOAC INTERNATIONAL 71, no. 5 (September 1, 1988): 945–49. http://dx.doi.org/10.1093/jaoac/71.5.945.

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Abstract The availability of antibody against deoxynivalenol (DON) triacetate (Tri-Ac-DON) has enabled development of a direct enzyme-linked immunosorbent assay (ELISA) and an indirect ELISA for DON in corn and wheat. In both assays, DON is extracted from the sample with acetonitrile- water, reacted with acetic anhydride to form Tri- Ac-DON, and diluted in phosphate buffer for analysis. Direct ELISA was found to be the more sensitive procedure. Fewer interferences are evidenced, and the assay is less time consuming than is indirect ELISA. For direct ELISA, recovery of 10-1000 ppb DON added to corn and wheat was 100% (SD 15, CV 15%) and 102.1% (SD 12.2, CV 11.9%), respectively. For indirect ELISA, overall recovery of 10- 1000 ppb DON added to wheat was 121.5% (SD 39.5, CV 32.5%); in the higher concentration range (500-1000 ppb), recovery was 105% (SD 18, CV 17%). The minimal detection level for DON was around 10 ppb. Analysis of 7 naturally contaminated samples for DON showed that the ELISA results agreed well with those obtained by radioimmunoassay and thin-layer chromatography.

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McKenna, S. L. B., D. C. Sockett, G. P. Keefe, J. McClure, J. A. VanLeeuwen, and H. W. Barkema. "Comparison of Two Enzyme-Linked Immunosorbent Assays for Diagnosis of Mycobacterium Avium Subsp. Paratuberculosis." Journal of Veterinary Diagnostic Investigation 17, no. 5 (September 2005): 463–66. http://dx.doi.org/10.1177/104063870501700510.

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Enzyme-linked immunosorbent assays (ELISAs) are used in Johne's disease (JD) control programs as a first screening for presence of the disease in a herd. A high sensitivity of the ELISA is therefore important, yet the commonly used ELISAs have relatively low sensitivity. The inclusion of an absorption phase, although improving specificity, potentially decreases sensitivity. Sera and feces of 383 adult dairy cows in 8 herds were used to compare the test characteristics of an absorbed and a nonabsorbed indirect ELISA for the detection of JD. The absorbed ELISA is based on a protoplasmic antigen, whereas the nonabsorbed uses a lipoarabinomannan-based antigen. The potential advantage of the nonabsorbed ELISA is that it may be less specific and more sensitive. Two herds certified free of JD were used to compare the specificity of the ELISAs. The other herds used to compare sensitivity were either infected with Mycobacterium avium subsp. paratuberculosis or had unknown status. Using fecal culture as a gold standard, the diagnostic specificity for the absorbed and nonabsorbed ELISAs were 98.4% and 87.9%, respectively. The diagnostic sensitivity was 72.4% and 65.5% for the absorbed and the nonabsorbed ELISA, respectively. Furthermore, a comparison using a fecal DNA probe as the comparison standard resulted in both ELISAs having a sensitivity of 61.9%. Agreement between the 2 ELISAs was moderate, with a kappa statistic of 0.58. The nonabsorbed ELISA did not have a higher sensitivity and had a lower specificity than the absorbed ELISA. Therefore, in this population, there was no advantage gained with using the nonabsorbed ELISA.

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HAZA, ANA I., PALOMA MORALES, ROSARIO MARTÍIN, TERESA GARCÍIA, GONZALO ANGUITA, ISABEL GONZÁLEZ, BERNABÉ SANZ, and PABLO E. HERNÁNDEZ. "Use of a Monoclonal Antibody and Two Enzyme-Linked Immunosorbent Assay Formats for Detection and Quantification of the Substitution of Caprine Milk for Ovine Milk." Journal of Food Protection 60, no. 8 (August 1, 1997): 973–77. http://dx.doi.org/10.4315/0362-028x-60.8.973.

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A stable hybridoma cell line (B2B) has been produced that secretes a monoclonal antibody (MAb) specific for goat's milk αS2-casein. The MAb B2B was used in two enzyme-linked immunosorbent assay (ELISA) formats for the detection and quantification of the presence of goat's milk in ewe's milk. In the indirect ELISA format the limit of detection was 0.5 to 15% (vol/vol) substitution of goat's milk for ewe's milk. Afterwards, a competitive indirect ELISA was successfully developed for the detection of 0.25 to 15% (vol/vol) of goat's milk in ewe's milk. This competitive indirect ELISA is a very sensitive assay; it can be performed in less than 5 h and is not influenced by the heat treatment of milk.

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Jabary, Osman M. "Detection of Brucella antibodies of sheep and goats by using two serological tests in Al-Sulaimanya governorate." Iraqi Journal of Veterinary Medicine 39, no. 2 (December 24, 2015): 32–37. http://dx.doi.org/10.30539/iraqijvm.v39i2.174.

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A research was carried out to detect Brucella antibodies of sheep and goats in Al-Sulaimanya governorate by using modified Rose Bengal test and indirect ELISA. A total of three hundred and eleven (311) whole blood samples (160 sheep and 151 goats) were collected randomly from eight different regions in Al-Sulaimanya governorate from unvaccinated flock with different ages. A total percentages of positive result by using modified Rose Bengal test was 14.46% (20% in sheep and 8.6% in goats), while by using indirect ELISA was 27.6% (35.2% in sheep and 19.2% in goats) with significant (P<0.05) differences. It revealed that rates to modified Rose Bengal test were 14.34% in female and 10.09% in males while to indirect ELISA 26.35% in female and 33.9% in males in sheep and goats, In conclusion of this study the high seroprevelence was at 1-3 years 19.2% and ˃3 years 33.96% according to modified Rose Bengal test and indirect ELISA, respectively.

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Paweska, Janusz T., Naazneen Moolla, Nadia Storm, Veerle Msimang, Ousman Conteh, Jacqueline Weyer, and Petrus Jansen van Vuren. "Evaluation of Diagnostic Performance of Three Indirect Enzyme-Linked Immunosorbent Assays for the Detection of IgG Antibodies to Ebola Virus in Human Sera." Viruses 11, no. 8 (July 24, 2019): 678. http://dx.doi.org/10.3390/v11080678.

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Filovirus serological diagnosis and epidemiological investigations are hampered due to the unavailability of validated immunoassays. Diagnostic performance of three indirect enzyme-linked immunosorbent assays (I-ELISA) was evaluated for the detection of IgG antibody to Ebola virus (EBOV) in human sera. One I-ELISA was based on a whole EBOV antigen (WAg) and two utilized recombinant nucleocapsid (NP) and glycoproteins (GP), respectively. Validation data sets derived from individual sera collected in South Africa (SA), representing an EBOV non-endemic country, and from sera collected during an Ebola disease (EBOD) outbreak in Sierra Leone (SL), were categorized according to the compounded results of the three I-ELISAs and real time reverse-transcription polymerase chain reaction (RT-PCR). At the cut-off values selected at 95% accuracy level by the two-graph receiver operating characteristic analysis, specificity in the SA EBOV negative serum panel (n = 273) ranged from 98.17% (GP ELISA) to 99.27% (WAg ELISA). Diagnostic specificity in the SL EBOV negative panel (n = 676) was 100% by the three ELISAs. The diagnostic sensitivity in 423 RT-PCR confirmed EBOD patients was dependent on the time when the serum was collected after onset of disease. It significantly increased 2 weeks post-onset, reaching 100% sensitivity by WAg and NP and 98.1% by GP I-ELISA.

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Fan, Titan S. L., Yi-Chun Xu, and Fun Sun Chu. "Simultaneous Analysis of T-2 Toxin and HT-2 Toxin by an Indirect Enzyme-Linked Immunosorbent Assay." Journal of AOAC INTERNATIONAL 70, no. 4 (July 1, 1987): 657–61. http://dx.doi.org/10.1093/jaoac/70.4.657.

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Abstract An indirect enzyme-linked immunosorbent assay (ELISA) for the detection of HT-2 toxin in the presence or absence of T-2 toxin is described. In the indirect ELISA, the relative cross-reactivities of antibodies against T-2 toxin (anti-T-2) with T-2 toxin and HT-2 toxin were 1 and 0.1, whereas anti-HT-2 cross-reactivities with T-2 toxin and HT-2 toxin were 0.33 and 1, respectively. Using such relationships, a formula was established that could be used to calculate the individual toxin concentration in a mixed sample after experimentally analyzing for T-2 and HT-2 toxins in the 2 indirect ELISAs. This method was tested by analyzing urine samples spiked with HT-2 toxin alone and samples spiked with both T-2 toxin and HT-2 toxin. A cleanup protocol for treatment of urine samples before ELISA was also established. The overall analytical recovery of HT-2 toxin when it was added at concentrations of 0.1-10 parts per billion (ppb) to the urine samples was ca 89%. When both T-2 and HT-2 toxins were added to the urine samples at equal concentrations of 0.5 to 5.0 ppb, their recoveries were 112 and 109%, respectively.

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Nawagitgul, Porntippa, Perry A. Harms, Igor Morozov, Brad J. Thacker, Steven D. Sorden, Chalermpol Lekcharoensuk, and Prem S. Paul. "Modified Indirect Porcine Circovirus (PCV) Type 2-Based and Recombinant Capsid Protein (ORF2)-Based Enzyme-Linked Immunosorbent Assays for Detection of Antibodies to PCV." Clinical and Vaccine Immunology 9, no. 1 (January 2002): 33–40. http://dx.doi.org/10.1128/cdli.9.1.33-40.2002.

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ABSTRACT Postweaning multisystemic wasting syndrome of swine associated with porcine circovirus (PCV) is a recently reported and economically important disease. Simple and reliable diagnostic methods are needed for detecting antibodies to PCV type 2 (PCV2) for monitoring of PCV infection. Here, we report the development of two modified indirect enzyme-linked immunosorbent assays (ELISAs): a PCV2 ELISA based on cell-culture-propagated PCV2 and an ORF2 ELISA based on recombinant major capsid protein. PCV2 and ORF2 ELISA detected antibodies to PCV2 and the capsid protein, respectively, in sera from pigs experimentally infected with PCV2 as early as 14 and 21 days postinoculation (dpi). The kinetics of the antibody response to PCV2 and the major capsid protein were similar. Repeatability tests revealed that the coefficients of variation of positive sera within and between runs for both assays were less than 30%. To validate the assays, PCV2 and ORF2 ELISAs were performed with 783 serum samples of young and adult pigs collected from different herds in the Midwestern United States and compared with an indirect immunofluorescent assay (IIF). Six out of 60 samples collected from nursery and growing pigs in 1987 were positive by both ELISA and IIF. Compared with IIF, the diagnostic sensitivity, specificity, and accuracy of PCV2 and ORF2 ELISAs were similar (>90%). The tests showed no cross-reactivity with antibodies to porcine parvovirus and porcine reproductive and respiratory syndrome virus. There was good agreement between the two ELISAs and between the ELISAs and IIF. The availability of the two ELISAs should accelerate our understanding of the host immune response to PCV2 and facilitate the development of prevention and control strategies by elucidating the ecology of PCV2 within swine populations.

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Hashim Handool Alfattli, Hams Hussien, and Saba Abood Ali Al-Mohamed. "First Serological and Molecular Detection of Leptospira interrogans serovar canicola Bacteria in Dogs in Some Iraqi Governorates." Journal of Education College Wasit University 1, no. 28 (August 6, 2017): 621–36. http://dx.doi.org/10.31185/eduj.vol1.iss28.28.

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The aim of present study was to serodetect of canine leptospirosis by using of indirect ELISA, and identification of Leptospira interrogans serovar canicola in serpositive dogs by application of PCR technique. For this purpose, 218 dogs from urban and rural regions related to three governorates were submitted for blood samples collection. The total results were revealed on 37/218 (16.97%) and 5/37 (13.51%) infected positive dogs by an indirect ELISA and PCR techniques, respectively. According to subjected study’s governorates, the positive results of indirect ELISA in Baghdad, Al-Qadisiyah and Dhi-Qar were 23/108 (21.3%), 10/79 (12.66%), and 4/31 (12.9%), respectively; while by PCR assay, the positive results {5/23 (21.74%)} had been detected in Baghdad only. Also, the relationship of positive dogs with some epidemiological risk factors has been discussed in this study. In association to inhabitant type, the rural and urban regions, respectively, were having 16/82 (19.51%) and 21/136 (15.44%) positive dogs by indirect ELISA; whereas, they have 4/16 (25%) and 1/21 (4.76%) positive dogs by PCR, respectively. In regarding to sex factor, the positive infected males and females, respectively, were amounted 13/67 (19.4%) and 24/151 (15.89%) by indirect ELISA; and 2/13 (15.38%) and 3/24 (12.5%) by PCR. In relation to age factor, >2years and £2 years groups have taken 36/149 (24.16%) and 1/69 (1.45%) positive dogs by indirect ELISA and 5/36 (13.89%) positive dogs for >2years group, only, by PCR assay. Statistically, the positive results were reported significant differences at level of P£0.05 between the study’s regions and between the groups related to each epidemiological risk factor.

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Ono, Erika, Manuel Mindlin Lafer, Lily Yin Weckx, Celso Granato, and Maria Isabel de Moraes-Pinto. "A simple and cheaper in house varicella zoster virus antibody indirect ELISA." Revista do Instituto de Medicina Tropical de São Paulo 46, no. 3 (June 2004): 165–68. http://dx.doi.org/10.1590/s0036-46652004000300008.

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We have developed a cheaper an simple in house indirect ELISA that uses the live attenuated VZV vaccine as a coating antigen. The alternative ELISA had an agreement of 94% when compared with a commercial VZV ELISA kit. Moreover, our ELISA proved to be more reliable than the kit when assessing true negative samples. By adding a standard serum, we were able to produce results in international units per millilitre. Also, the addition of an extra step with 8M urea allowed the assessment of VZV IgG avidity without excessive costs. The cost per sample to test VZV IgG was 2.7 times cheaper with our ELISA, allowing the testing of many samples without the burden of production of VZV antigen in the laboratory.

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Naqvi, Muhammad Ali-ul-Husnain, Sana Zahra Naqvi, Muhammad Ali Memon, Kalibixiati Aimulajiang, Muhammad Haseeb, Lixin Xu, Xiaokai Song, Xiangrui Li, and Ruofeng Yan. "Combined Use of Indirect ELISA and Western Blotting with Recombinant Hepatocellular Carcinoma-Associated Antigen 59 Is a Potential Immunodiagnostic Tool for the Detection of Prepatent Haemonchus contortus Infection in Goat." Animals 9, no. 8 (August 13, 2019): 548. http://dx.doi.org/10.3390/ani9080548.

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Haemonchus contortus is recognized as one of the important health problems in small ruminants, leading to reduced production and economic loss for farmers worldwide. Prepatent diagnosis of H. contortus infection is crucial to improve control strategies as this helminth may remove up to one-fifth of total erythrocytes and may cause anemia, edema, diarrhea, and ultimately death in young animals. In this study, one of the excretory and secretory products, rHc-HCA59, was purified and used as antigen to detect specific antibodies in H. contortus infected goats during prepatent stage of infection using indirect enzyme linked immunosorbent assay (ELISA) as screening test. All goats (n = 38) were housed indoor, experimentally infected with 8000 infective larvae (L3) of H. contortus, and serum samples were collected prior to infection and at 14th day of infection. Immunoblotting was performed to confirm the results of indirect ELISA, evaluate the cross reactivity against rHc-HCA59 in sera of most common co-infecting parasites and rectify the false negative samples. Furthermore, three different batches of rHc-HCA59 were produced to evaluate the repeatability of ELISA. No eggs were detected in feces of all goats collected at 7th and 14th day of infection but, H. contortus eggs were detected at 21 days post infection in the feces. Indirect ELISA performed in this study showed 87% sensitivity and 100% specificity. The western blot analysis confirmed immunoreactivity in serum samples which scored positive in indirect ELISA and recognized the samples as negative which had OD450 lower than negative cut-off value in indirect ELISA. Furthermore, all false negative sera (n = 5) that had OD450 value between positive and negative cut-off value in rHc-HCA59 based ELISA were clearly positive in western blot. Moreover, no cross-reactivity was detected in ELISA and western blotting against rHc-HCA59 in positive sera of Toxoplasma gondii, Fasciola hepatica, and Trichinella spiralis. The results of this study concluded that combined use of indirect ELISA and western blotting with rHc-HCA59 is a potential immunodiagnostic tool for the detection of H. contortus infection during prepatent period in goats.

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Patel, D., W. Egner, D. Gleeson, G. Wild, and A. Ward. "Detection of serum M2 anti-mitochondrial antibodies by enzyme-linked immunosorbent assay is potentially less specific than by immunofluorescence." Annals of Clinical Biochemistry: International Journal of Laboratory Medicine 39, no. 3 (May 1, 2002): 304–7. http://dx.doi.org/10.1258/0004563021902008.

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Aim To compare the predictive values of enzyme-linked immunosorbent assays (ELISAs) and indirect immunofluorescence (IIF) techniques for the detection of M2 anti-mitochondrial antibodies. Methods Commercial ELISAs are widely available for the detection of antimitochondrial antibody subtypes in primary biliary cirrhosis (PBC). We compared the results from two ELISAs (one recombinant, one purified antigen) with those from two IIF methods in a well-defined cohort of PBC patients and in patients with systemic lupus erythematosus, Sjögren's syndrome, sicca syndrome, systemic sclerosis, rheumatoid arthritis and blood donor controls. Results There was good correlation between a rodent substrate IIF and ELISA A ( r = 0·9134), but poor correlation with ELISA B ( r = 0·5999), which produced many false-positive results in the control population. We show that rodent IIF alone or human epithelial cell (HEp-2000®) screening with confirmation by ELISA produce similar predictive values for PBC and require lesser degrees of skilled interpretation of IIF patterns. Conclusions We conclude that the specificities of IIF are greater than the ELISA methods (99% versus 85-97%), although the ELISAs are slightly more sensitive in biopsy-proven PBC. Careful in-house validation of all new ELISA technologies is mandatory for good laboratory practice, but IIF in experienced hands remains an effective and specific assay.

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Journal articles on the topic 'Indirect ELISA'

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