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Journal articles on the topic 'Denaturing high performance liquid chromatography (DHPLC)'

Author: Grafiati

Published: 21 February 2023

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1

Cole, David E. C., Francisco H. J. Yun, Betty Y. L. Wong, Andrew Y. Shuen, Ronald A. Booth, Alfredo Scillitani, Svetlana Pidasheva, Xiang Zhou, Lucie Canaff, and Geoffrey N. Hendy. "Calcium-sensing receptor mutations and denaturing high performance liquid chromatography." Journal of Molecular Endocrinology 42, no. 4 (January 29, 2009): 331–39. http://dx.doi.org/10.1677/jme-08-0164.

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The calcium-sensing receptor (CASR), a plasma membrane G-protein-coupled receptor, is expressed in parathyroid gland and kidney, and controls systemic calcium homeostasis. Inactivating CASR mutations are associated with familial hypocalciuric hypercalcemia (FHH) and neonatal severe hyperparathyroidism, and activating mutations cause autosomal dominant hypocalcemia (ADH). CASR mutation identification plays an important role in the clinical management of mineral metabolism disorders. We describe here a high-throughput method using screening with denaturing high performance liquid chromatography (DHPLC) to initially interrogate 12 amplicons covering translated exons and exon/intron boundaries, followed by sequencing of any amplicon with a modified melting curve relative to wild type, and direct sequencing of a 13th amplicon encoding the COOH-terminal tail to distinguish causative mutations from three common missense single nucleotide polymorphisms. A blinded analysis of 32 positive controls representing mutations throughout the CASR sequence, as well as 22 negative controls, yielded a concordance rate of 100%. We report eight novel and five recurrent FHH mutations, along with six novel and two recurrent ADH mutations. Thus, DHPLC provides a rapid and effective means to screen for CASR mutations.

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Xu, Li, Jason Evans, Thomas Ling, Kathy Nye, and Peter Hawkey. "Rapid Genotyping of CTX-M Extended-Spectrum β-Lactamases by Denaturing High-Performance Liquid Chromatography." Antimicrobial Agents and Chemotherapy 51, no. 4 (January 8, 2007): 1446–54. http://dx.doi.org/10.1128/aac.01088-06.

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ABSTRACT Denaturing high-performance liquid chromatography (dHPLC) is a powerful technique which has been used extensively to detect genetic variation. This is the first report of the application of dHPLC for rapid genotyping of bacterial β-lactamase genes. The technique was specifically developed to genotype members of all bla CTX-M DNA homology groups. Thirteen well-defined bla CTX-M extended-spectrum β-lactamase (ESBL)-producing strains were used to develop and optimize the dHPLC genotyping assay. Further evaluation was carried out with a blinded panel of 62 clinical isolates. The results of bla CTX-M genotyping achieved by dHPLC were comparable to the typing results obtained by DNA sequencing. Applying the newly developed dHPLC-based genotyping method, we successfully genotyped all 73 bla CTX-M ESBL-producing strains from the 4-month survey study. Furthermore, we found the first reported cases in the United Kingdom of clinically significant disease caused by CTX-M-14- and CTX-M-1-producing Escherichia coli strains. We conclude that the novel dHPLC assay is highly accurate, rapid, and cost-effective for the genotyping of bla CTX-M-producing ESBLs and has great potential for determining the clinical relevance of different and new bla CTX-M genotypes, as well as for epidemiological studies and surveillance programs.

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Crépin, Michel, Pascal Pigny, Fabienne Escande, Catherine Cardot Bauters, Alain Calender, Sylvain Lefevre, Marie-Pierre Buisine, Nicole Porchet, and Marie-Françoise Odou. "Evaluation of denaturing high performance liquid chromatography for the mutational analysis of the MEN1 gene." Journal of Molecular Endocrinology 36, no. 2 (April 2006): 369–76. http://dx.doi.org/10.1677/jme.1.01903.

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The identification of mutations in the MEN1 gene causing MEN1 has represented a challenge since the cloning of the gene in 1997 because of the lack of mutation hot-spots in the gene and the lack of phenotype–genotype correlations. The use of denaturing high performance liquid chromatography (DHPLC), a high throughput, reliable and automated heteroduplex-based technique, is the ideal for mutation detection in MEN1. In this work, DHPLC was optimised for the screening of the nine coding exons and splice junctions of MEN1. Thanks to collaboration between two French laboratories recognised as reference centres for genotypic MEN1 diagnosis (Lyon and Lille), a blind retrospective study conducted in a cohort of 160 unrelated MEN1 probands with (or without) known germline mutations was undertaken to evaluate the sensitivity of DHPLC. We were able to detect 101 different sequence variations by DHPLC, distributed in the 10 analysed DNA fragments and corresponding to 100% of mutation detection compared with direct sequencing. 1·2% of samples were considered as false positive, exhibiting a heterogenous profile. DHPLC did not detect five cases of deletion or duplication of complete exons, neither did direct sequencing, showing the limits of the technique. Nevertheless, the method appeared to allow automated, rapid and low-cost mutation detection with high accuracy. Direct sequencing can be then applied to identify the sequence variations on the targeted DNA fragments showing heterozygous profile by DHPLC. In conclusion, genotypic diagnosis of MEN1 can benefit from DHPLC in terms of efficacy, rapidity and cost.

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Priha, Outi, Mari Nyyssönen, Malin Bomberg, Arja Laitila, Jaakko Simell, Anu Kapanen, and Riikka Juvonen. "Application of Denaturing High-Performance Liquid Chromatography for Monitoring Sulfate-Reducing Bacteria in Oil Fields." Applied and Environmental Microbiology 79, no. 17 (June 21, 2013): 5186–96. http://dx.doi.org/10.1128/aem.01015-13.

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ABSTRACTSulfate-reducing bacteria (SRB) participate in microbially induced corrosion (MIC) of equipment and H2S-driven reservoir souring in oil field sites. Successful management of industrial processes requires methods that allow robust monitoring of microbial communities. This study investigated the applicability of denaturing high-performance liquid chromatography (DHPLC) targeting the dissimilatory sulfite reductase ß-subunit (dsrB) gene for monitoring SRB communities in oil field samples from the North Sea, the United States, and Brazil. Fifteen of the 28 screened samples gave a positive result in real-time PCR assays, containing 9 × 101to 6 × 105dsrBgene copies ml−1. DHPLC and denaturing gradient gel electrophoresis (DGGE) community profiles of the PCR-positive samples shared an overall similarity; both methods revealed the same samples to have the lowest and highest diversity. The SRB communities were diverse, and differentdsrBcompositions were detected at different geographical locations. The identifieddsrBgene sequences belonged to several phylogenetic groups, such asDesulfovibrio,Desulfococcus,Desulfomicrobium,Desulfobulbus,Desulfotignum,Desulfonatronovibrio, andDesulfonauticus. DHPLC showed an advantage over DGGE in that the community profiles were very reproducible from run to run, and the resolved gene fragments could be collected using an automated fraction collector and sequenced without a further purification step. DGGE, on the other hand, included casting of gradient gels, and several rounds of rerunning, excising, and reamplification of bands were needed for successful sequencing. In summary, DHPLC proved to be a suitable tool for routine monitoring of the diversity of SRB communities in oil field samples.

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Kota, Raja, Markus Wolf, Wolfgang Michalek, and Andreas Graner. "Application of denaturing high-performance liquid chromatography for mapping of single nucleotide polymorphisms in barley (Hordeum vulgare L.)." Genome 44, no. 4 (August 1, 2001): 523–28. http://dx.doi.org/10.1139/g01-053.

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Recent advances in DNA sequence analysis and the establishment of high-throughput assays have provided the framework for large-scale discovery and analysis of DNA sequence variation. In this context, single nucleotide polymorphisms (SNPs) are of particular interest. To initiate a systematic approach to develop an SNP map of barley (Hordeum vulgare L.), we have employed denaturing high-performance liquid chromatography (DHPLC) to analyse segregating SNP patterns in a doubled-haploid (DH) mapping population. To this end, SNPs between the parental genotypes were identified using a direct sequencing approach. Once a SNP was established between the parents, the optimal melting temperature of the PCR fragment containing the SNP was predicted for its analysis by DHPLC. Following the detection of the optimal temperature, the DH lines were analysed for the presence of either of the alleles. To test the utility of the analysis, data from previously mapped RFLP markers from which these SNPs were derived were compared. Results from these experiments indicate that DHPLC can be efficiently employed in analysing SNPs on a high-throughput scale.Key words: denaturing high performance liquid chromatography, doubled-haploid lines, restriction fragment length polymorphism, genetic mapping, molecular markers.

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Wagner, Andreas Otto, Cornelia Malin, and Paul Illmer. "Application of Denaturing High-Performance Liquid Chromatography in Microbial Ecology: Fermentor Sludge, Compost, and Soil Community Profiling." Applied and Environmental Microbiology 75, no. 4 (December 16, 2008): 956–64. http://dx.doi.org/10.1128/aem.01411-08.

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ABSTRACT Genetic fingerprinting methods, such as denaturing gradient gel electrophoresis (DGGE), are used in microbial ecology for the analysis of mixed microbial communities but are associated with various problems. In the present study we used a new alternative method: denaturing high-performance liquid chromatography (dHPLC). This method was previously shown to work with samples from water and gut flora but had not yet been applied to complex environmental samples. In contrast to other publications dealing with dHPLC, we used a commonly available HPLC system. Samples from different origins (fermentor sludge, compost, and soil), all ecologically significant, were tested, and the 16S rRNA gene was amplified via PCR. After optimization of the HPLC elution conditions, amplicons of pure cultures and mixed microbial populations could be separated successfully. Systematic differentiation was carried out by a cloning approach, since fraction collection of the peaks did not result in satisfactory fragment separation. dHPLC was evaluated as a tool for microbial community analysis on a genetic level and demonstrated major improvements compared to gel-based fingerprinting methods, such as DGGE, that are commonly used in microbial ecology.

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Troedsson, Christofer, Richard F. Lee, Vivica Stokes, Tina L. Walters, Paolo Simonelli, and Marc E. Frischer. "Development of a Denaturing High-Performance Liquid Chromatography Method for Detection of Protist Parasites of Metazoans." Applied and Environmental Microbiology 74, no. 14 (May 23, 2008): 4336–45. http://dx.doi.org/10.1128/aem.02131-07.

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ABSTRACT Increasingly, diseases of marine organisms are recognized as significant biotic factors affecting ecosystem health. However, the responsible disease agents are often unknown and the discovery and description of novel parasites most often rely on morphological descriptions made by highly trained specialists. Here, we describe a new approach for parasite discovery, utilizing denaturing high-performance liquid chromatography (DHPLC) reverse-phase ion-paring technology. Systematic investigations of major DHPLC variables, including temperature, gradient conditions, and target amplicon characteristics were conducted to develop a mechanistic understanding of DNA fragment separation by DHPLC. As a model system, 18S rRNA genes from the blue crab (Callinectes sapidus) and a parasitic dinoflagellate Hematodinium sp. were used. Binding of 18S rRNA gene PCR amplicons to the DNA separation column in the presence of triethylammonium acetate (TEAA) was inversely correlated with temperature and could be predicted based on the estimated DNA helicity of the PCR amplicon. Amplicons of up to 498 bp were resolved as single chromatographic peaks if they had high (>95%) DNA helicity. Amplicons that differed by as few as 2 bp could be resolved. Separation of 18S rRNA gene PCR amplicons was optimized by simultaneous manipulation of both temperature and solvent gradients. The optimal conditions included targeting regions of high DNA helicity (>95%), temperatures in the range of 57 to 63°C, and a linear acetonitrile gradient from 13.75 to 17.5% acetonitrile in 0.1 M TEAA (55 to 70% buffer B) over a 9-min period. Under these conditions, amplicons from a variety of parasites and their hosts can be separated and detected by DHPLC.

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Kim, Il-Jin, Yong Shin, Hio Chung Kang, Jae-Hyun Park, Ja-Lok Ku, Hye-Won Park, Hye-Rin Park, et al. "Robust microsatellite instability (MSI) analysis by denaturing high-performance liquid chromatography (DHPLC)." Journal of Human Genetics 48, no. 10 (September 12, 2003): 525–30. http://dx.doi.org/10.1007/s10038-003-0070-y.

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Ray, Pierre F., Paule Benit, Jean Paul Bonnefont, and Arnold Munnich. "Chronology of reported denaturing high performance liquid chromatography (DHPLC)-based prenatal diagnoses." Prenatal Diagnosis 23, no. 1 (January 2003): 81. http://dx.doi.org/10.1002/pd.496.

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Bénit, Paule, Jean-Paul Bonnefont, Ali Kara Mostefa, Christine Francannet, Arnold Munnich, and Pierre F. Ray. "Denaturing high-performance liquid chromatography (DHPLC)-based prenatal diagnosis for tuberous sclerosis." Prenatal Diagnosis 21, no. 4 (March 7, 2001): 279–83. http://dx.doi.org/10.1002/pd.55.

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Agulnik, J. S., V. Cohen, J. Jarry, G. Batist, D. Small, H. Kreisman, N. A. Tejada, G. Chong, and W. H. Miller. "Use of denaturing high performance liquid chromatography (dHPLC) for detection of EGFR mutations in patients with NSCLC." Journal of Clinical Oncology 24, no. 18_suppl (June 20, 2006): 10068. http://dx.doi.org/10.1200/jco.2006.24.18_suppl.10068.

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10068 Background: Somatic mutations of the epidermal growth factor receptor (EGFR) gene may predict responsiveness to tyrosine kinase inhibitors in patients with non-small cell lung cancer (NSCLC). These mutations are commonly identified using DNA sequencing methods. Although considered the gold standard, this approach is expensive and time-consuming. In addition, a high ratio of tumour to normal tissue DNA is required for optimal results which is not often available in biopsies obtained from these patients. The primary objective of this study was to develop a rapid and sensitive method for screening of EGFR mutations. Materials and Methods: Clinical specimens from 110 NSCLC patients were analysed for EGFR mutations in exons 19 and 21. After DNA extraction and PCR, both dHPLC and direct sequencing were performed and results were compared. Results: Sequencing revealed a total of 7 (6%) mutations: 4 missense mutations in exon 21 and 3 deletion mutations in exon 19. dHPLC showed the presence of genomic alterations in 18 (16%) samples, including the 7 identified by sequencing plus 11 additional samples (10 in exon 19 and one in exon 21). dHPLC fractions were isolated, reamplified, and sequenced to confirm these results. In serial dilution studies, dHPLC was able to detect mutations in samples containing as little as 10% mutated DNA whereas direct sequencing required at least 30%. Conclusions: dHPLC is an efficient and accurate, as well as a a more sensitive method for screening of genomic alterations in exons 19 and 21 of the EGFR gene compared to direct sequencing. This data suggests that dHPLC should be implemented as a screening tool for detection of EGFR mutations. No significant financial relationships to disclose.

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Flex, Elisabetta, Alessandro De Luca, Maria Rosaria D'Apice, Anna Buccino, Bruno Dallapiccola, and Giuseppe Novelli. "Rapid scanning of myotubularin (MTM1) gene by denaturing high-performance liquid chromatography (DHPLC)." Neuromuscular Disorders 12, no. 5 (June 2002): 501–5. http://dx.doi.org/10.1016/s0960-8966(01)00328-5.

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Sutomo, Retno, Sunartini Hapsara, Suryono Yudha Patria, and Hajime Nakamura. "Detection of the jaundice-related G71R mutation in the UGT1A1 gene by denaturing high performance liquid chromatography (DHPLC)." Paediatrica Indonesiana 49, no. 1 (March 1, 2009): 1. http://dx.doi.org/10.14238/pi49.1.2009.1-6.

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Background The G71R mutation in the UGT1A1 gene has beenassociated with neonatal jaundice and other cases of hereditary,unconjugated hyperbilirubinemia in several Asian populations.Currently, DNA sequencing is the only method available toidentify the mutation, which can be time- and labor-intensive,particularly for such projects as population-based genetic studies.A relatively new method, denaturing high performance liquidchromatography (DHPLC), is increasingly used to detect variousmutations.Objective The aim of the present study was to investigate theability of DHPLC to detect the G71R mutation, in comparisonwith the gold standard of sequencing analysis.Methods Seventy-two infants were enrolled. Following genomicDNA extraction, exon 1 of the UGT1A1 gene was amplified bypolymerase chain reaction (PCR). Afterwards, the G71R mutationwas simultaneously, and blindly, determined in all subjects byDHPLC and sequence analysis. The performance of the DHPLCanalysis, compared to the sequence analysis, was assessed in termsof sensitivity and specificity.Results DHPLC detected the G71 R mutation in 31 individuals.Of these, 26 were heterozygous and 5 were homozygous for themutation. This method did not find the mutation in 41 otherindividuals. Sequence analysis produced identical results for allindividuals.Conclusion DHPLC analysis is capable of detecting the G71Rmutation in the UGT1A1 with a degree of sensitivity andspecificity (100% each) that is comparable to sequencing analysis.

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HUNG, CHIA-CHENG, SHU-CHIN CHIEN, YI-NING SU, JIMMY PS CHERN, KAI-HSIN LIN, and WIN-LI LIN. "DENATURING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY: AN EFFICIENT SCREENING APPROACH IN THE GENETIC DIAGNOSIS OF HEMOGLOBIN HAMMERSMITH." Biomedical Engineering: Applications, Basis and Communications 18, no. 06 (December 25, 2006): 343–47. http://dx.doi.org/10.4015/s1016237206000506.

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In this study we here present the first report of the detection of the rare beta-thalassemia mutation in the Taiwanese population: Hemoglobin Hammersmith (Hb Hammersmith). The mutation was detected by denaturing high-performance liquid chromatography (DHPLC) screening followed by automated direct sequencing. The mutation was found in an affected woman and her immature female fetus in the heterozygous state. Molecular abnormality of Hb Hammersmith results from an abnormal beta chain with an amino acid substitution (condon 42, TTT→TCT, Phe→Ser) in the beta-globin (HBB) gene with the clinical presentation of hemolytic anemia. Given known wide spectrum of beta-thalassemia alleles in the Taiwanese population, the present report further confirmed the high heterogeneity rate. This result indicated that the importance of the efficient screening approach by DHPLC for genetic diagnosis in beta-thalassemia.

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Taylor, Matthew R. G., Misi L. Robinson, and Luisa Mestroni. "Analysis of Genetic Variations of Lamin A/C Gene (LMNA) by Denaturing High-Performance Liquid Chromatography." Journal of Biomolecular Screening 9, no. 7 (October 2004): 625–28. http://dx.doi.org/10.1177/1087057104266393.

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The human LMNA gene, when mutated, has been shown to cause at least 7 human diseases: dilated cardiomyopathy, Emery Dreifuss muscular dystrophy, limb girdle muscular dystrophy, familial partial lipodystrophy, Charcot Marie tooth disease type II, mandibuloacral dysplasia, and Hutchinson-Gilford Progeria (OMIM #176670). This article describes a high-throughput method for screening the human lamin A/C ( LMNA) gene for genetic mutations and sequence variation using denaturing high-performance liquid chromatography (DHPLC). In the present study, 76 patients with dilated cardiomyopathy were screened for mutations using DHPLC and sequence analysis. Abnormal elution profiles were identified and sequenced on an ABI 377 automatic sequencer. Heterozygous LMNA mutations were detected in 8% of the affected patients. In addition, a number of intronic and exonic single nucleotide polymorphisms were identified. LMNA mutations are clinically relevant in at least 6 human diseases. This study provides a protocol for high-throughput LMNA analysis applicable both in the research and in the clinical diagnostic setting.

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Canu, Annie, Ahmed Abbas, Brigitte Malbruny, François Sichel, and Roland Leclercq. "Denaturing High-Performance Liquid Chromatography Detection of Ribosomal Mutations Conferring Macrolide Resistance in Gram-Positive Cocci." Antimicrobial Agents and Chemotherapy 48, no. 1 (January 2004): 297–304. http://dx.doi.org/10.1128/aac.48.1.297-304.2004.

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ABSTRACT Mutations in genes coding for L4 (rplD) or L22 (rplV) ribosomal proteins or in 23S rRNA (rrl gene) are reported as a cause of macrolide resistance in streptococci and staphylococci. This study was aimed at evaluating a denaturing high-performance liquid chromatography (DHPLC) technique as a rapid mutation screening method. Portions of these genes were amplified by PCR from total DNA of 48 strains of Streptococcus pneumoniae (n = 22), Staphylococcus aureus (n = 16), Streptococcus pyogenes (n = 6), Streptococcus oralis (n = 2), and group G streptococcus (n = 2). Thirty-seven of these strains were resistant to macrolides and harbored one or several mutations in one or two of the target genes, and 11 were susceptible. PCR products were analyzed by DHPLC. All mutations were detected, except a point mutation in a pneumococcal rplD gene. The method detected one mutated rrl copy out of six in S. aureus. This automated method is promising for screening of mutations involved in macrolide resistance in gram-positive cocci.

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Wolford, Johanna, Conrad Ballecer, Dalia Blunt, and Michal Prochazka. "High-throughput SNP detection by using DNA pooling and denaturing high performance liquid chromatography (DHPLC)." Human Genetics 107, no. 5 (December 1, 2000): 483–87. http://dx.doi.org/10.1007/s004390000396.

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Franciosa, Giovanna, Manoocheher Pourshaban, Alessandro De Luca, Anna Buccino, Bruno Dallapiccola, and Paolo Aureli. "Identification of Type A, B, E, and F Botulinum Neurotoxin Genes and of Botulinum Neurotoxigenic Clostridia by Denaturing High-Performance Liquid Chromatography." Applied and Environmental Microbiology 70, no. 7 (July 2004): 4170–76. http://dx.doi.org/10.1128/aem.70.7.4170-4176.2004.

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ABSTRACT Denaturing high-performance liquid chromatography (DHPLC) is a recently developed technique for rapid screening of nucleotide polymorphisms in PCR products. We used this technique for the identification of type A, B, E, and F botulinum neurotoxin genes. PCR products amplified from a conserved region of the type A, B, E, and F botulinum toxin genes from Clostridium botulinum, neurotoxigenic C. butyricum type E, and C. baratii type F strains were subjected to both DHPLC analysis and sequencing. Unique DHPLC peak profiles were obtained with each different type of botulinum toxin gene fragment, consistent with nucleotide differences observed in the related sequences. We then evaluated the ability of this technique to identify botulinal neurotoxigenic organisms at the genus and species level. A specific short region of the 16S rRNA gene which contains genus-specific and in some cases species-specific heterogeneity was amplified from botulinum neurotoxigenic clostridia and from different food-borne pathogens and subjected to DHPLC analysis. Different peak profiles were obtained for each genus and species, demonstrating that the technique could be a reliable alternative to sequencing for the rapid identification of food-borne pathogens, specifically of botulinal neurotoxigenic clostridia most frequently implicated in human botulism.

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Troedsson, Christofer, Richard F. Lee, Tina Walters, Vivica Stokes, Karrie Brinkley, Verena Naegele, and Marc E. Frischer. "Detection and Discovery of Crustacean Parasites in Blue Crabs (Callinectes sapidus) by Using 18S rRNA Gene-Targeted Denaturing High-Performance Liquid Chromatography." Applied and Environmental Microbiology 74, no. 14 (May 23, 2008): 4346–53. http://dx.doi.org/10.1128/aem.02132-07.

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ABSTRACT Recently, we described a novel denaturing high-performance liquid chromatography (DHPLC) approach useful for initial detection and identification of crustacean parasites. Because this approach utilizes general primers targeted to conserved regions of the 18S rRNA gene, a priori genetic sequence information on eukaryotic parasites is not required. This distinction provides a significant advantage over specifically targeted PCR assays that do not allow for the detection of unknown or unsuspected parasites. However, initial field evaluations of the DHPLC assay suggested that because of PCR-biased amplification of dominant host genes it was not possible to detect relatively rare parasite genes in infected crab tissue. Here, we describe the use of a peptide nucleic acid (PNA) PCR hybridization blocking probe in association with DHPLC (PNA-PCR DHPLC) to overcome inherent PCR bias associated with amplification of rare target genes by use of generic primers. This approach was utilized to detect infection of blue crabs (Callinectes sapidus) by the parasitic dinoflagellate Hematodinium sp. Evaluation of 76 crabs caught in Wassaw Sound, GA, indicated a 97% correspondence between detection of the parasite by use of a specific PCR diagnostic assay and that by use of PNA-PCR DHPLC. During these studies, we discovered one crab with an association with a previously undescribed protist symbiont. Phylogenetic analysis of the amplified symbiont 18S rRNA gene indicated that it is most closely related to the free-living kinetoplastid parasite Procryptobia sorokini. To our knowledge, this is the first report of this parasite group in a decapod crab and of this organism exhibiting a presumably parasitic life history.

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Liu, W., D. I. Smith, K. J. Rechtzigel, S. N. Thibodeau, and C. D. James. "Denaturing high performance liquid chromatography (DHPLC) used in the detection of germline and somatic mutations." Nucleic Acids Research 26, no. 6 (March 1, 1998): 1396–400. http://dx.doi.org/10.1093/nar/26.6.1396.

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Pigullo, Simona, Alessandro De Luca, Paolo Barone, Roberta Marchese, Emilia Bellone, Alessia Colosimo, Cesa Scaglione, et al. "Mutational analysis of parkin gene by denaturing high-performance liquid chromatography (DHPLC) in essential tremor." Parkinsonism & Related Disorders 10, no. 6 (August 2004): 357–62. http://dx.doi.org/10.1016/j.parkreldis.2004.04.012.

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Lam, Ching-Wan. "First application of denaturing high-performance liquid chromatography (DHPLC) for prenatal diagnosis of genetic disease." Prenatal Diagnosis 22, no. 1 (January 2002): 79–80. http://dx.doi.org/10.1002/pd.230.

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Lin, Shin-Yu, Shu-Chin Chien, Yi-Ning Su, Chien-Nan Lee, and Chih-Ping Chen. "Rapid genetic analysis of oculocutaneous albinism (OCA1) using denaturing high performance liquid chromatography (DHPLC) system." Prenatal Diagnosis 26, no. 5 (2006): 466–70. http://dx.doi.org/10.1002/pd.1439.

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Lin, Doris, Jayne A. Goldstein, Anand N. Mhatre, Lawrence R. Lustig, Markus Pfister, and Anil K. Lalwani. "Assessment of denaturing high-performance liquid chromatography (DHPLC) in screening for mutations in connexin 26 (GJB2)." Human Mutation 18, no. 1 (2001): 42–51. http://dx.doi.org/10.1002/humu.1148.

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Magliozzi, Monia, Maria Piane, Isabella Torrente, Lorenzo Sinibaldi, Giovanni Rizzo, Camilla Savio, Patrizia Lulli, Alessandro De Luca, Bruno Dallapiccola, and Luciana Chessa. "DHPLC Screening ofATMGene in Italian Patients Affected by Ataxia-Telangiectasia: Fourteen NovelATMMutations." Disease Markers 22, no. 4 (2006): 257–64. http://dx.doi.org/10.1155/2006/740493.

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The gene for ataxia-telangiectasia (A-T:MIM:#208900), ATM, spans about 150~kb of genomic DNA and is composed of 62 coding exons. ATM mutations are found along the entire coding sequence of the gene, without evidence of mutational hot spots. Using DNA as the starting material, we used denaturing high performance liquid chromatography (DHPLC) technique to search for ATM gene mutations. Initially, DHPLC was validated in a retrospective study of 16 positive control samples that included 19 known mutations; 100% of mutations were detected. Subsequently, DHPLC was used to screen for mutations a cohort of 22 patients with the classical form of A-T. A total of 27 different mutations were identified on 38 of the 44 alleles, corresponding to a 86% detection rate. Fourteen of the mutations were novel. In addition, 15 different variants and polymorphisms of unknown functional significance were found. The high incidence of new and individual A-T mutations in our cohort of patients demonstrates marked mutational heterogeneity of A-T in Italy and corroborate the efficiency of DHPLC as a method for the mutation screening of A-T patients.

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Ribeiro, Diogo, Ana Cardoso, Ana Joana Duarte, Luis Vieira, and Olga Amaral. "Efficient IDUA Gene Mutation Detection with Combined Use of dHPLC and Dried Blood Samples." ISRN Molecular Biology 2013 (April 22, 2013): 1–4. http://dx.doi.org/10.1155/2013/451298.

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Objectives. Development of a simple mutation directed method in order to allow lowering the cost of mutation testing using an easily obtainable biological material. Assessment of the feasibility of such method was tested using a GC-rich amplicon. Design and Methods. A method of denaturing high-performance liquid chromatography (dHPLC) was improved and implemented as a technique for the detection of variants in exon 9 of the IDUA gene. The optimized method was tested in 500 genomic DNA samples obtained from dried blood spots (DBS). Results. With this dHPLC approach it was possible to detect different variants, including the common p.Trp402Ter mutation in the IDUA gene. The high GC content did not interfere with the resolution and reliability of this technique, and discrimination of G-C transversions was also achieved. Conclusion. This PCR-based dHPLC method is proved to be a rapid, a sensitive, and an excellent option for screening numerous samples obtained from DBS. Furthermore, it resulted in the consistent detection of clearly distinguishable profiles of the common p.Trp402Ter IDUA mutation with an advantageous balance of cost and technical requirements.

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Herbert, O., M. Trossaert, P. Boisseau, E. Fressinaud, and F. Gerson. "Evaluation of denaturing high-performance liquid chromatography (DHPLC) in the screening of mutations in hemophilia B patients." Journal of Thrombosis and Haemostasis 2, no. 12 (December 2004): 2267–69. http://dx.doi.org/10.1111/j.1538-7836.2004.01019.x.

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Bernasconi, Luca, Roberto Herklotz, Martin Hergersberg, and Andreas Huber. "dHPLC as a Rapid and Reliable Screening Method for A-Thalassemia Screening." Blood 104, no. 11 (November 16, 2004): 3774. http://dx.doi.org/10.1182/blood.v104.11.3774.3774.

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Abstract Background: a-thalassemia, the most prevalent of all thalassemias, is an inherited single gene disorder of the hemoglobin synthesis characterized by the absence or the reduced expression of a-globin genes resulting in an imbalanced ratio between a- and b-gobin chain synthesis. While some DNA alterations lead to microcytic anemia, the most others cause severe anemia and intrauterine death if co-inherited in a homozygous constellation. The most common genetic defects leading to the a-thalassemia phenotype are deletions of one or more of the two highly homologous a-globin genes. Less frequently, a-thalassemia derives from nondeletional mutations located in parts of the a-globin gene that are critical for its normal expression. So far the identification of the genetic background of a-thalassemic patients has included specific amplification of each a-globin gene followed by laborious and expensive mutation analysis methods such as denaturing gradient gel electrophoresis (DGGE), single-strand conformation polymorphism (SSCP) or direct sequencing of the entire a-genes. Method: We attempted to evaluate the performance of denaturing high performance liquid chromatography (dHPLC) technique to identify nondeletional mutations located in the a1- or a2-globin gene. Due to the high sequence homology of a1- and a2-globin genes, separate amplification of the two genes followed by nested PCR were performed. This generated four overlapping amplicons (a-dHPLC1 to 4) covering the whole a1/a2-globin locus including the 3′- and 5′-untranslated regions. The formation of DNA hetero-/homoduplexes was ensured by denaturing and slow reannealing of crude PCR products. Through a DNA separation column under partial-denaturing conditions, DNA homoduplexes were retained longer than their corresponding heteroduplexes generating distinguishable chromatographic profiles and hence allowing for the differentiation between wildtype and mutant DNA. Samples which differed from the wildtype in their elution profile were sequenced in both directions. Results: 50 patients showing a presumptive a-thalassemic hematological profile not due to deletional mutations (excluded by gap PCR technique) were tested. 10 carriers of a-thalassemia nondeletional alleles (previously identified by endonuclease restriction analysis) and 10 healthy patients were also included in the evaluation. Conclusion: Our results show that dHPLC is a reliable, sensitive, and specific screening method to detect pointmutations and small deletions located in the a-globin genes. Furthermore, the low costs of analysis and the rapidity of the screening (about 6 min/sample) make the dHPLC technique an appropriate alternative to technically demanding and time consuming mutation screening methods such as DGGE or SSCP analysis.

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Betz, Beate, Andrea R. Florl, Hans-Helge Seifert, Peter Dall, Wolfgang A. Schulz, and Dieter Niederacher. "Denaturing high-performance liquid chromatography (DHPLC) as a reliable high-throughput prescreening method for aberrant promoter methylation in cancer." Human Mutation 23, no. 6 (2004): 612–20. http://dx.doi.org/10.1002/humu.20033.

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Shinka, T., T. Naroda, T. Tamura, K. Sasahara, and Y. Nakahori. "A rapid and simple method for sex identification by heteroduplex analysis, using denaturing high-performance liquid chromatography (DHPLC)." Journal of Human Genetics 46, no. 5 (April 2001): 263–66. http://dx.doi.org/10.1007/s100380170076.

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Deng, Da-jun, Guo-ren Deng, Jing Zhou, and Hui-jun Xin. "Detection of CPG methylations in human mismatch repair gene hMLH1 promoter by denaturing high-performance liquid chromatography (DHPLC)." Chinese Journal of Cancer Research 12, no. 3 (September 2000): 171. http://dx.doi.org/10.1007/bf02983460.

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Chien, Shu-Chin, Chien-Nan Lee, Chia-Cheng Hung, Po-Nien Tsao, Yi-Ning Su, and Fon-Jou Hsieh. "Rapid prenatal diagnosis of X-linked chronic granulomatous disease using a denaturing high-performance liquid chromatography (DHPLC) system." Prenatal Diagnosis 23, no. 13 (2003): 1092–96. http://dx.doi.org/10.1002/pd.761.

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Benigni, Michele, Stefania Battistini, and Claudia Ricci. "Genotyping of Macrophage Migration Inhibitory Factor (MIF) CATT5–8 Repeat Polymorphism by Denaturing High-Performance Liquid Chromatography (DHPLC)." Molecular Biotechnology 54, no. 3 (December 15, 2012): 874–79. http://dx.doi.org/10.1007/s12033-012-9636-2.

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Piepoli, Ada, Enrico Schirru, Angela Mastrorilli, Annamaria Gentile, Rosa Cotugno, Michele Quitadamo, Antonio Merla, Mauro Congia, Paolo Usai Satta, and Francesco Perri. "Genotyping of the Lactase-Phlorizin Hydrolase C/T-13910 Polymorphism by Means of a New Rapid Denaturing High-Performance Liquid Chromatography-Based Assay in Healthy Subjects and Colorectal Cancer Patients." Journal of Biomolecular Screening 12, no. 5 (August 2007): 733–39. http://dx.doi.org/10.1177/1087057107301328.

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Adult-type hypolactasia results from the progressive decline of lactase-phlorizin hydrolase activity in enterocytes after weaning. Lactase nonpersistence may determine a primary lactose intolerance with reduced diary product consumption, which is possibly related to an increased risk of colon cancer. Recently, a genetic variant C/T—13910 upstream of the lactase-phlorizin hydrolase ( LCT) gene has been strongly correlated with the lactase persistence/nonpersistence trait in both family and case-control studies. The authors validate a denaturing high-performance liquid chromatography (dHPLC)—based assay versus conventional genotype sequencing in detecting the C/T—13910 polymorphism of LCT and evaluate its prevalence in 2 different Italian geographical areas and in colorectal cancer patients. DNA samples of 157 healthy subjects and 124 colon cancer patients from Apulia and of 97 healthy subjects from Sardinia were evaluated for the C/T—13910 polymorphism by dHPLC, sequencing, and restriction fragment length polymorphism (RFLP). Under optimized conditions, dHPLC was as sensitive as DNA sequencing and detected a new genetic variant (T/C-13913) in 2 individuals that was not identified by RFLP assay. Frequency of lactase nonpersistence genotype (C/C—13910) was similar in healthy subjects from 2 different Italian geographical areas and not increased in patients with colorectal cancer. The results indicate that the dHPLC method may be used as a rapid, noninvasive, and laborsaving screening tool for genotyping C/T—13910 polymorphism, with high success, low cost, and reproducibility. ( Journal of Biomolecular Screening 2007:733-739)

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Edelman, E. Jennifer, Yelena Maksimova, Feride Duru, Cigdem Altay, and Patrick G. Gallagher. "A complex splicing defect associated with homozygous ankyrin-deficient hereditary spherocytosis." Blood 109, no. 12 (June 15, 2007): 5491–93. http://dx.doi.org/10.1182/blood-2006-09-046573.

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Abstract Defects in erythrocyte ankyrin are the most common cause of typical, dominant hereditary spherocytosis (HS). Detection of ankyrin gene mutations has been complicated by allelic heterogeneity, large gene size, frequent de novo mutations, and associated mRNA instability. Using denaturing high-performance liquid chromatography (DHPLC)–based mutation detection, a mutation in the splice acceptor of exon 17 was discovered in a Turkish family. Reticulocyte RNA and functional minigene splicing assays in heterologous cells revealed that this mutation was associated with a complex pattern of aberrant splicing, suggesting that removal of intron 16 is important for ordered ankyrin mRNA splicing. As predicted by clinical, laboratory, and biochemical studies, the parents were heterozygous and the proband was homozygous for this mutation. These data indicate that DHPLC offers a highly sensitive, economic, and rapid method for mutation detection and, unlike previously suggested, homozygosity for a mutation associated with dominant ankyrin-linked HS may be compatible with life.

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Sagarzazu, Noelia Isabel, Maribel Martínez, Cristina Algarra, Javier Butrón, Carlos J. González-Navarro, and Raquel Virto. "Optimization of denaturing high performance liquid chromatography technique for rapid detection and identification of acetic acid bacteria of interest in vinegar production." Acetic Acid Bacteria 2, no. 1s (February 26, 2013): 5. http://dx.doi.org/10.4081/aab.2013.s1.e5.

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This paper evaluates the use of denaturing high performance liquid chromatography (DHPLC) technology for the discrimination of genetic differences in the 16S rRNA and alcohol dehydrogenase (AdhA) genes among bacterial species based on its efficiency and sensitivity to enable the detection and discrimination of different genetic sequences. In order to optimize DHPLC protocols for the analysis of 16S rRNA gene fragments amplified from bacteria, DNA isolated from 22 different strains representing main bacterial groups of interest in food microbiology was analyzed. While the use of 16S rRNA gene did not allow to difference two wild strains of <em>Acetobacter malorum</em>, this region revealed as useful to differentiate them from some pathogenic bacteria as <em>Escherichia coli</em>, <em>Salmonella typhimurium</em>, <em>Listeria monocytogenes</em>, <em>Listeria innocua</em>, <em>Clostridium perfringens </em>or <em>Sthapylococcus aureus</em>, from spoilage microorganisms as <em>Xantomonas vesicatoria</em> and <em>Alicyclobacillus</em> spp., and also from lactic acid bacteria as <em>Lactobacillus plantarum</em>, <em>Lactobacillus casei</em>, <em>Lactobacillus sakei</em>, <em>Lactobacillus acidophilus</em>, <em>Streptococcus thermophilus </em>and <em>Lactococcus lactis</em> that may suppose technological risk during vinegar production. The results demonstrate that 16S rRNA gene region is not adequate for the discrimination of the acetic acid bacteria (AAB) strains, so AdhA gene was selected to identify the two wild strains of <em>Acetobacter malorum</em>. Also 6 different reference strains of AAB were separated based on differences in AdhA gene region. DHPLC technology is able to discriminate between these two wild strains of <em>A. malorum</em> based on differences existing in the AdhA gene region. The data obtained indicate that the technique is capable of identifying most bacteria at species level and even at strain level with optimization of the protocols. This is of particular relevance in the case of AAB due to their poor recovery on culture media and difficulties in detection of viable but non cultivable cells.

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Cho, Sang-Man. "Development of a Denaturing High-Performance Liquid Chromatography (DHPLC) Assay to Detect Parasite Infection in Grass Shrimp Palaemonetes pugio." Fisheries and aquatic sciences 15, no. 2 (June 30, 2012): 107–15. http://dx.doi.org/10.5657/fas.2012.0107.

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38

Charon, C. M., and C. T. Dolphin. "Sequence variation in the human flavin-containing monooxygenase 3 gene (FMO3): identification by denaturing-high performance liquid chromatography (DHPLC)." Biochemical Society Transactions 28, no. 5 (October 1, 2000): A435. http://dx.doi.org/10.1042/bst028a435.

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39

Han, Song, David Cooper, and Meena Upadhyaya. "Evaluation of denaturing high performance liquid chromatography (DHPLC) for the mutational analysis of the neurofibromatosis type 1 ( NF1 ) gene." Human Genetics 109, no. 5 (November 1, 2001): 487–97. http://dx.doi.org/10.1007/s004390100594.

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40

Nelken, Joanna, Reza Meshkani, Nita Chahal, Brian McCrindle, and Khosrow Adeli. "Detection of familial defective apoB (FDB) mutations in hypercholesterolemic children and adolescents by denaturing high performance liquid chromatography (DHPLC)." Clinical Biochemistry 41, no. 6 (April 2008): 395–99. http://dx.doi.org/10.1016/j.clinbiochem.2007.12.018.

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41

Lu, T., and D. B. Oerther. "A NOVEL STUDY OF MICROBIAL COMMUNITY RESPONSE TO TOXIC SHOCK LOADINGS BY DENATURING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY (DHPLC) METHOD." Proceedings of the Water Environment Federation 2007, no. 13 (January 1, 2007): 5001–9. http://dx.doi.org/10.2175/193864707787969568.

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42

Koumaravelou, K., Z. Shoaib, C. Adithan, D. Charron, A. Srivastava, R. Tamouza, and R. Krishnamoorthy. "Analysis of human glutathione S-transferase alpha 1 (hGSTA1) gene promoter polymorphism using Denaturing High Performance Liquid Chromatography (DHPLC)." Clinica Chimica Acta 412, no. 15-16 (July 2011): 1465–68. http://dx.doi.org/10.1016/j.cca.2011.04.019.

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43

Berginc, Gašper, and Damjan Glavač. "Rapid and Accurate Approach for Screening of Microsatellite Unstable Tumours Using Quasimonomorphic Mononucleotide Repeats and Denaturating High Performance Liquid Chromatography (DHPLC)." Disease Markers 26, no. 1 (2009): 19–26. http://dx.doi.org/10.1155/2009/901532.

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MSI analysis is becoming increasingly important for the detection of both hereditary non-polyposis colorectal cancer and sporadic primary colorectal tumours with MSI high phenotype. The Bethesda panel of five microsatellite markers has been proposed to provide uniform criteria for MSI analysis. Here we report on an MSI analysis approach using quasimonomorphic mononucleotide repeats and denaturating high performance liquid chromatography (DHPLC). We analysed 595 newly diagnosed colorectal tumours and 145 normal samples. Microsatellite markers BAT-25, BAT-26, NR-21, NR-22, and NR-27 were amplified in multiplex reaction and analysed using DHPLC and capillary electrophoresis (CE). DHPLC conditions for analysis of MSI multiplex assay were evaluated and tested. Analysis and cross-examination of the results obtained from 96 samples using DHPLC and capillary electrophoresis showed the same sensitivity and specificity of the two approaches for detecting MSI-H tumours. Using our new approach we showed that the tested markers are quasimonomorphic in a Slovenian population, with frequencies of polymorphisms 0.07%, 1.4%, 2.1%, 1.4%, and 1.4% for BAT-25, BAT-26, NR-21, NR-22, and NR-27, respectively. Forty-three (7.2%) new MSI-H tumours were identified, of which 84% showed instability in all 5 tested markers. Overall, we developed a high-throughput, robust, accurate and cost-effective approach for the detection of MSI-H tumours.

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Hecksel, Kathleen A., Gordon W. Dewald, and David P. Steensma. "Ferrochelatase Mutation Screening by Denaturing High Performance Liquid Chomatography in Idiopathic Acquired Sideroblastic Anemia (Refractory Anemia with Ringed Sideroblasts) and Erythrocytic Protoporphyria." Blood 106, no. 11 (November 16, 2005): 3735. http://dx.doi.org/10.1182/blood.v106.11.3735.3735.

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Abstract BACKGROUND: The molecular etiology of idiopathic acquired sideroblastic anemia (IASA), now considered a form of myelodysplastic syndrome, is currently unknown. Romslo et al (Blood 1982) reported a patient with IASA who had moderately elevated free erythrocytic protoporphyrin (FEP); this observation is now frequently made in IASA, helping distinguish IASA (high FEP) from inherited sideroblastosis (low FEP). In addition, rare patients with erythropoietic protoporphyria (EPP)—defined by cutaneous photosensitivity, dramatically elevated FEP levels, and germline point mutations in the ferrochelatase (FECH) gene at 18q21.3—have ringed sideroblasts in their marrow. We hypothesized that patients with IASA might have acquired somatic FECH mutations. METHODS AND RESULTS: We designed a denaturing high performance liquid chromatography (DHPLC) assay to explore this possibility and to avoid problems related to mutation screening in the setting of mixed clonality. To validate the DHPLC assay, the coding region of the FECH gene from 2 molecularly undiagnosed EPP patients without liver disease was analyzed. In one patient, both a heterozygous 69delG mutation (GenBank Accession NM_000140) and heterozygosity for the FECH expression-modulating IVS3-48C/T polymorphism were detected. In the other patient, no FECH coding mutations or polymorphisms were detected, either by DHPLC or conventional dye sequencing. FEP measurements were then obtained on 2 IASA patients and were elevated (65 and 115 mcg/dL; normal 1–10 mcg/dL) with normal urine and fecal porphyrins. Genomic DNA obtained from these 2 patients as well as archival DNA from 30 other patients with IASA was amplified and tested for FECH mutations. No coding region mutations were detected. Synonymous polymorphisms in exon 7 (rs536765) and 9 (rs536560) were found in 6/32 (19%; normal heterozygosity 0.398) and 14/32 (44%; normal heterozygosity 0.402) samples, respectively. The IVS3-48C/T polymorphism (rs2272783) was found in 3/32 (9%) (prevalence in the general population is 11%, Gouya Nat Genet 2002). In addition, 3 intronic polymorphisms not in the refSNP database were detected: IVS8+34 C/T (2/32), IVS8-61delG (5/32), and IVS9-59delA (2/32). CONCLUSION: IASA is not associated with coding mutations in FECH. Elevation of FEP in ASA must instead be due to the lack of mitochondrial iron in the proper form for incorporation into the porphyrin ring; attention should instead focus on factors responsible for maintaining the appropriate redox state and compartmentalization of iron. DHPLC can detect FECH mutations and polymorphisms, but because of the high frequency of the latter and the consequent need to sequence multiple exons, it is not a practical screening tool for studying undiagnosed EPP patients.

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Koyama, Renata Guedes, Rosa M. R. P. S. Castro, Marco Túlio De Mello, Sergio Tufik, and Mario Pedrazzoli. "Simple Detection of Large InDeLS by DHPLC: The ACE Gene as a Model." Journal of Biomedicine and Biotechnology 2008 (2008): 1–5. http://dx.doi.org/10.1155/2008/562183.

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Insertion-deletion polymorphism (InDeL) is the second most frequent type of genetic variation in the human genome. For the detection of large InDeLs, researchers usually resort to either PCR gel analysis or RFLP, but these are time consuming and dependent on human interpretation. Therefore, a more efficient method for genotyping this kind of genetic variation is needed. In this report, we describe a method that can detect large InDeLs by DHPLC (denaturating high-performance liquid chromatography) using the angiotensin-converting enzyme (ACE) gene I/D polymorphism as a model. The InDeL targeted in this study is characterized by a 288 bp Alu element insertion (I). We used DHPLC at nondenaturating conditions to analyze the PCR product with a flow through the chromatographic column under two different gradients based on the differences between D and I sequences. The analysis described is quick and easy, making this technique a suitable and efficient means for DHPLC users to screen InDeLs in genetic epidemiological studies.

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Gupta, Ruchi, Rajni Gaind, Laishram Chandreshwor Singh, Bianca Paglietti, Monorama Deb, Salvatore Rubino, John Wain, and Seemi Farhat Basir. "Detection of mutations in gyrB using denaturing high performance liquid chromatography (DHPLC) among Salmonella enterica serovar Typhi and Paratyphi A." Transactions of The Royal Society of Tropical Medicine and Hygiene 110, no. 12 (December 2016): 684–89. http://dx.doi.org/10.1093/trstmh/trx002.

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Roberts, P. S., S. Jozwiak, D. J. Kwiatkowski, and S. L. Dabora. "Denaturing high-performance liquid chromatography (DHPLC) is a highly sensitive, semi-automated method for identifying mutations in the TSC1 gene." Journal of Biochemical and Biophysical Methods 47, no. 1-2 (January 2001): 33–37. http://dx.doi.org/10.1016/s0165-022x(00)00149-4.

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Sahli, Chaima Abdelhafidh, Ikbel Ben Salem, Latifa Jouini, Naouel Laouini, Rym Dabboubi, Sondes Hadj Fredj, Hajer Siala, et al. "Setup of a Protocol of Molecular Diagnosis of β-Thalassemia Mutations in Tunisia using Denaturing High-Performance Liquid Chromatography (DHPLC)." Journal of Clinical Laboratory Analysis 30, no. 5 (April 18, 2016): 392–98. http://dx.doi.org/10.1002/jcla.21867.

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Hutzler, M., E. Geiger, and F. Jacob. "Use of PCR-DHPLC (Polymerase Chain Reaction-Denaturing High Performance Liquid Chromatography) for the Rapid Differentiation of IndustrialSaccharomyces pastorianusandSaccharomyces cerevisiaeStrains." Journal of the Institute of Brewing 116, no. 4 (2010): 464–74. http://dx.doi.org/10.1002/j.2050-0416.2010.tb00798.x.

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Mounier, Jérôme, Audrey Gouëllo, Marlène Keravec, Solène Le Gal, Grégory Pacini, Stella Debaets, Gilles Nevez, Gilles Rault, Georges Barbier, and Geneviève Héry-Arnaud. "Use of denaturing high-performance liquid chromatography (DHPLC) to characterize the bacterial and fungal airway microbiota of cystic fibrosis patients." Journal of Microbiology 52, no. 4 (February 17, 2014): 307–14. http://dx.doi.org/10.1007/s12275-014-3425-5.

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