title: Rapid detection of HLA-B*51 by real-time polymerase chain reaction and high-resolution melting analysis.
abstract: HLA-B*51, a class I human leukocyte antigen (HLA) molecule, is the strongest known genetic risk factor for Behçet disease. However, there are only few articles reporting methods to determine the presence or absence of HLA-B51. For this reason, we designed and developed an easy, fast, and inexpensive real-time high-resolution melting (HRM) assay to detect HLA-B*51. We genotyped 61 samples by our HRM assay and by conventional polymerase chain reaction, and no discrepancies were found between results. Besides, a subgroup of 25 samples was also genotyped in a different laboratory, and another subgroup of 16 samples was obtained from the International Histocompatibility Working Group DNA Bank, and a full concordance of results was observed with those obtained by HRM. Regarding the identifying system evaluated, we obtained 100% of specificity, sensibility, and repeatability, and 0% of false positive and false negative rates. Therefore, this HRM analysis is easily applicable to the rapid detection of HLA-B*51, exhibits a high speed, and requires a very low budget.
authors.affiliation: Genètica Molecular-Laboratori Clínic, Hospital Universitari de Bellvitge, L'Hospitalet de Llobregat, Catalonia, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Imperiali
authors.affiliation: Genètica Molecular-Laboratori Clínic, Hospital Universitari de Bellvitge, L'Hospitalet de Llobregat, Catalonia, Spain.
authors.forename: P
authors.initials: P
authors.lastname: Alía-Ramos
authors.affiliation: Genètica Molecular-Laboratori Clínic, Hospital Universitari de Bellvitge, L'Hospitalet de Llobregat, Catalonia, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Padró-Miquel
mesh_terms: D001528:Behcet Syndrome / Q000175:diagnosis* / Q000235:genetics / Q000276:immunology; D001681:Biological Assay / Q000191:economics / Q000295:instrumentation / Q000592:standards*; D017931:DNA Primers / Q000737:chemistry*; D005838:Genotype; D060005:Genotyping Techniques / Q000191:economics / Q000295:instrumentation / Q000592:standards*; D059949:HLA-B51 Antigen / Q000235:genetics* / Q000276:immunology; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000191:economics / Q000295:instrumentation / Q000592:standards*; D009691:Nucleic Acid Denaturation; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D017931:DNA Primers; D059949:HLA-B51 Antigen
keywords: Behçet disease; HLA-B*51; high-resolution melting assay; method; real-time PCR