title: Comparison of methods for in-house screening of HLA-B*57:01 to prevent abacavir hypersensitivity in HIV-1 care.
abstract: Abacavir is a nucleoside reverse transcriptase inhibitor used as part of combination antiretroviral therapy in HIV-1-infected patients. Because this drug can cause a hypersensitivity reaction that is correlated with the presence of the HLA-B*57:01 allotype, screening for the presence of HLA-B*57:01 is recommended before abacavir initiation. Different genetic assays have been developed for HLA-B*57:01 screening, each with specific sensitivity, turnaround time and assay costs. Here, a new real-time PCR (qPCR) based analysis is described and compared to sequence specific primer PCR with capillary electrophoresis (SSP PCR CE) on 149 patient-derived samples, using sequence specific oligonucleotide hybridization combined with high resolution SSP PCR as gold standard. In addition to these PCR based methods, a complementary approach was developed using flow cytometry with an HLA-B17 specific monoclonal antibody as a pre-screening assay to diminish the number of samples for genetic testing. All three assays had a maximum sensitivity of >99. However, differences in specificity were recorded, i.e. 84.3%, 97.2% and >99% for flow cytometry, qPCR and SSP PCR CE respectively. Our data indicate that the most specific and sensitive of the compared methods is the SSP PCR CE. Flow cytometry pre-screening can substantially decrease the number of genetic tests for HLA-B*57:01 typing in a clinical setting.
authors: Ghent University, Department of Internal Medicine, Ghent, Belgium.
authors: Ward
authors: W
authors: De Spiegelaere
authors: Ghent University, Department of Clinical Chemistry, Microbiology and Immunology, Ghent, Belgium.
authors: Jan
authors: J
authors: Philippé
authors: Ghent University, Department of Internal Medicine, Ghent, Belgium.
authors: Karen
authors: K
authors: Vervisch
authors: Ghent University, Department of Clinical Chemistry, Microbiology and Immunology, Ghent, Belgium.
authors: Chris
authors: C
authors: Verhofstede
authors: Ghent University, Department of Internal Medicine, Ghent, Belgium.
authors: Eva
authors: E
authors: Malatinkova
authors: Ghent University, Department of Internal Medicine, Ghent, Belgium.
authors: Maja
authors: M
authors: Kiselinova
authors: Ghent University, Department of Internal Medicine, Ghent, Belgium.
authors: Wim
authors: W
authors: Trypsteen
authors: Ghent University, Department of Internal Medicine, Ghent, Belgium.
authors: Pawel
authors: P
authors: Bonczkowski
authors: Ghent University, Department of Internal Medicine, Ghent, Belgium.
authors: Dirk
authors: D
authors: Vogelaers
authors: Ghent University, Department of Internal Medicine, Ghent, Belgium.
authors: Steven
authors: S
authors: Callens
authors: Université Catholique de Louvain, IREC, AIDS Reference Laboratory, Brussels, Belgium.
authors: Jean
authors: J
authors: Ruelle
authors: Saint-Pierre University Hospital, Department of Infectious Diseases, Bruxelles, Belgium.
authors: Kabamba
authors: K
authors: Kabeya
authors: Saint-Pierre University Hospital, Department of Infectious Diseases, Bruxelles, Belgium.
authors: Stephane
authors: S
authors: De Wit
authors: Ghent University, Center for Medical Genetics, Ghent, Belgium.
authors: Petra
authors: P
authors: Van Acker
authors: HILA, Laboratory for Histocompatibility & Immunogenetics Red Cross Flanders, Mechelen, Belgium.
authors: Vicky
authors: V
authors: Van Sandt
authors: HILA, Laboratory for Histocompatibility & Immunogenetics Red Cross Flanders, Mechelen, Belgium.
authors: Marie-Paule
authors: MP
authors: Emonds
authors: Ghent University, Center for Medical Genetics, Ghent, Belgium.
authors: Paul
authors: P
authors: Coucke
authors: Ghent University, Department of Internal Medicine, Ghent, Belgium.
authors: Erica
authors: E
authors: Sermijn
authors: Ghent University, Department of Internal Medicine, Ghent, Belgium.
authors: Linos
authors: L
authors: Vandekerckhove
mesh_terms: D000483:Alleles; D023241:Antiretroviral Therapy, Highly Active; D017931:DNA Primers / Q000138:chemical synthesis / Q000235:genetics; D015224:Dideoxynucleosides / Q000008:administration & dosage* / Q000009:adverse effects; D004342:Drug Hypersensitivity / Q000209:etiology / Q000235:genetics / Q000276:immunology / Q000517:prevention & control*; D019075:Electrophoresis, Capillary; D005434:Flow Cytometry; D015870:Gene Expression; D005820:Genetic Testing / Q000379:methods; D015658:HIV Infections / Q000188:drug therapy* / Q000235:genetics / Q000276:immunology / Q000821:virology; D015497:HIV-1 / Q000187:drug effects / Q000201:enzymology / Q000235:genetics; D015235:HLA-B Antigens / Q000235:genetics* / Q000276:immunology; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D018894:Reverse Transcriptase Inhibitors / Q000008:administration & dosage* / Q000009:adverse effects; D012680:Sensitivity and Specificity; D000099227:Abacavir
chemical_list: D017931:DNA Primers; D015224:Dideoxynucleosides; D015235:HLA-B Antigens; D018894:Reverse Transcriptase Inhibitors; D000099227:Abacavir; C484830:HLA-B*57:01 antigen; C037449:HLA-B17 antigen