title: Comparative Analysis of Real-Time Polymerase Chain Reaction Methods to Typing HLA-B*57:01 in HIV-1-Positive Patients.
abstract: The HLA-B*57:01 allele is strongly associated with the hypersensitivity reaction to Abacavir (ABC). Therefore, treatment guidelines recommend that patients initiating ABC are preventively tested for the presence of this allele. To date, four different commercial assays based on the real-time quantitative polymerase chain reaction (Q-PCR) technique are available for the detection of HLA-B*57:01: Duplicα-RealTime Reagent Set HLA-B*57:01 by Euroclone, HLA-B*57:01 Real-TM by Sacace Biotechnologies, COBAS AmpliPrep/COBAS TaqMan HLA-B*57:01 Screening Test by Roche Diagnostic, and HLA-B*57:01 by Nuclear Laser Medicine. The study was carried out to compare the performance of the first three commercially available Q-PCR kits in a routine clinical setting. A total of 98 samples from Policlinico Umberto I Hospital were tested. Results obtained by the Duplicα-RealTime Genotyping kit and AmpliPrep/TaqMan system were 100% concordant. In contrast, genotyping by the HLA-B*57:01 Real-TM kit showed poor agreement with the other systems, that is, 12 out of 33 positive samples were detected as HLA-B*57:01 negative. To confirm the correct genotype of these discordant samples, two additional methods with rapid turnaround times and already implemented into routine clinical practice were used, that is, a PCR-based microsequence-specific primer DNA typing test and a laboratory-developed screening test in Q-PCR. All 12 discordant samples were genotyped as HLA-B*57:01-positive samples using these two additional methods in a single-blinded manner, thus confirming the low sensitivity of HLA-B*57:01 Real-TM test. These findings underline the need to compare results obtained with commercial assays before choosing a test suitable for use in a routine clinical laboratory.
authors.affiliation: 1 Department of Molecular Medicine, Sapienza University of Rome , Rome, Italy .
authors.forename: Francesca
authors.initials: F
authors.lastname: Falasca
authors.affiliation: 2 Institute of Pharmacology, Catholic University Medical School , Rome, Italy .
authors.forename: Cinzia
authors.initials: C
authors.lastname: Dello Russo
authors.affiliation: 3 Department of Experimental Medicine, Sapienza University of Rome , Rome, Italy .
authors.forename: Barbara
authors.initials: B
authors.lastname: Mora
authors.affiliation: 4 ViiV Healthcare , Verona, Italy .
authors.forename: Antonella
authors.initials: A
authors.lastname: Pirazzoli
authors.affiliation: 5 Department of Clinical Medicine, Sapienza University of Rome , Rome, Italy .
authors.forename: Alessandra
authors.initials: A
authors.lastname: Fantauzzi
authors.affiliation: 2 Institute of Pharmacology, Catholic University Medical School , Rome, Italy .
authors.forename: Pierluigi
authors.initials: P
authors.lastname: Navarra
authors.affiliation: 3 Department of Experimental Medicine, Sapienza University of Rome , Rome, Italy .
authors.forename: Antonio
authors.initials: A
authors.lastname: Pizzuti
authors.affiliation: 6 Department of Public Health and Infectious Diseases, Sapienza University of Rome , Rome, Italy .
authors.forename: Corrado
authors.initials: C
authors.lastname: De Vito
authors.affiliation: 1 Department of Molecular Medicine, Sapienza University of Rome , Rome, Italy .
authors.forename: Guido
authors.initials: G
authors.lastname: Antonelli
authors.affiliation: 1 Department of Molecular Medicine, Sapienza University of Rome , Rome, Italy .
authors.forename: Ombretta
authors.initials: O
authors.lastname: Turriziani
mesh_terms: D019380:Anti-HIV Agents / Q000009:adverse effects*; D015224:Dideoxynucleosides / Q000009:adverse effects*; D064420:Drug-Related Side Effects and Adverse Reactions / Q000517:prevention & control*; D005260:Female; D060005:Genotyping Techniques / Q000379:methods*; D015658:HIV Infections / Q000188:drug therapy*; D015235:HLA-B Antigens / Q000235:genetics*; D006801:Humans; D008297:Male; D008875:Middle Aged; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D000099227:Abacavir
chemical_list: D019380:Anti-HIV Agents; D015224:Dideoxynucleosides; D015235:HLA-B Antigens; D000099227:Abacavir; C103707:HLA-B57 antigen