HLA-B*5701 typing: evaluation of an allele-specific polymerase chain reaction melting assay.
Inheritance of HLA-B*5701 is a strong predictor of a hypersensitivity reaction to the anti-HIV drug abacavir. The identification of susceptible individuals prior to the institution of abacavir therapy is therefore of clinical importance and has generated demand for a simple and rapid diagnostic test for carriage of HLA-B*5701. In this study, we describe the development of such a method based on allele-specific polymerase chain reaction (AS-PCR) and melting curve analysis. Ninety-six patient samples including 36 HLA-B*5701-positive samples and 60 HLA-B*5701-negative samples were analysed. Compared with sequence-based typing, this method had 100% sensitivity and specificity for the HLA-B*5701 allele. In conclusion, the AS-PCR/melting curve approach minimises post-polymerase chain reaction handling processing and provides an attractive alternative to currently described AS-PCR methods.
Hammond
E
E
Centre for Clinical Immunology and Biomedical Statistics, Royal Perth Hospital and Murdoch University, Wellington Street, Western Australia 6000, Australia. e.hammond@murdoch.edu.au
Mamotte
C
C
Nolan
D
D
Mallal
S
S
D000483:Alleles; D019380:Anti-HIV Agents / Q000009:adverse effects / Q000627:therapeutic use; D001483:Base Sequence; D001681:Biological Assay; D015224:Dideoxynucleosides / Q000494:pharmacology; D004342:Drug Hypersensitivity / Q000235:genetics / Q000517:prevention & control; D005820:Genetic Testing; D015235:HLA-B Antigens / Q000235:genetics*; D006650:Histocompatibility Testing / Q000379:methods*; D006358:Hot Temperature; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D018894:Reverse Transcriptase Inhibitors / Q000494:pharmacology; D012680:Sensitivity and Specificity; D000099227:Abacavir
D019380:Anti-HIV Agents; D015224:Dideoxynucleosides; D015235:HLA-B Antigens; D018894:Reverse Transcriptase Inhibitors; D000099227:Abacavir; C484830:HLA-B*57:01 antigen