Novel Sensitive, Specific and Rapid Pharmacogenomic test for the Prediction of Abacavir Hypersensitivity Reaction: HLA-B*57:01 Detection by Real-Time PCR

Abstract

We provided a detailed characterization of a novel HLA-B*57:01 screening test, which can be easily implemented by those laboratories already involved in the detection of viral load and virus genotyping. Original submitted 26 October 2010; Revision submitted 13 December 2010.

Prevalence of Human Leukocyte Antigen-B*5701 Among HIV-infected Children in Thailand and Cambodia

“…13,14 One of the obstacles for screening for HLA-B*5701 in developing countries is laboratory cost. Low cost techniques, such as polymerase chain reaction sequence-specific primer-based genotyping, may make screening feasible in resource-limited settings 15 Whether or not HLA-B*5701 screening has been performed, clinical training on early detection of the clinical symptoms of abacavir hypersensitivity, especially in the first 6 weeks of treatment, should be emphasized. Abacavir should be immediately discontinued in any patient who develops clinical signs and symptoms consistent with hypersensitivity and rechallenge is contraindicated.…”

Development of multiplex pyrosequencing for HLA-B*57:01 screening using single nucleotide polymorphism haplotype

“…The Prospective Random Evaluation of DNA Screening in a Clinical Trial (PREDICT‐1) has shown that ABC HSR is strongly associated with HLA‐B*57:01 . Such a finding has been confirmed by various studies, which lead to the implementation of recommendation guidelines by the US Food and Drug Administration (FDA) and international HIV treatment guidelines for HLA‐B*57:01 screening before initiating treatment with ABC …”

Comparison of Methods for In-House Screening of HLA-B*57:01 to Prevent Abacavir Hypersensitivity in HIV-1 Care

“…The qPCR was based on the protocol described by Dello-Russo et al, that was adapted to avoid the necessity of the initial HLA-B PCR [ 16 ]. This assay targets the exon 2 (Primers 193F, 319R and HLA 57:01 exon 2 probe; S2 Table ) and exon 3 (Primers 345F, 419R and HLAB57:01 exon 3 probe; S2 Table ) region of the HLA-B*57 : 01 allele.…”

“…This method has been further optimized by subsequent capillary electrophoresis, enhancing the sensitivity of the assay [ 15 ]. More recently, new assays were developed for HLA-B*57 : 01 typing on a qPCR platform [ 11 , 16 , 17 ]. This enables detection of primer specificity through differentiating Cq values by SYBR Green quantitative (q)PCR [ 16 ], or analysis of allele specific PCR by high resolution melting [ 17 ].…”

“…More recently, new assays were developed for HLA-B*57 : 01 typing on a qPCR platform [ 11 , 16 , 17 ]. This enables detection of primer specificity through differentiating Cq values by SYBR Green quantitative (q)PCR [ 16 ], or analysis of allele specific PCR by high resolution melting [ 17 ]. Implementation of these assays on a qPCR platform significantly decreases the processing and reaction time as well as reagent costs [ 11 ].…”