4.0 Article

Rapid Near Point-of-Care Assay for HLA-B*57:01 Genotype Associated with Severe Hypersensitivity Reaction to Abacavir

Journal

AIDS RESEARCH AND HUMAN RETROVIRUSES
Volume 37, Issue 12, Pages 930-935

Publisher

MARY ANN LIEBERT, INC
DOI: 10.1089/aid.2021.0103

Keywords

oligonucleotide ligation assay; HLA-B*57; 01; abacavir; hypersensitivity reactions; pediatric ART; genetic screening

Funding

  1. National Institutes of Health (NIH) [R01 AI110375, R01AI145486]
  2. Clinical and Retrovirology Research Core and the Molecular Profiling and Computational Biology Core of the University of Washington Fred Hutch Center for AIDS Research [P30 AI027757]
  3. International Ma-ternal Pediatric Adolescent AIDS Clinical Trials Network(IMPAACT) [UM1AI068632, UM1AI068616, UM1AI106716]
  4. National Institute of Allergy and Infectious Diseases (NIAID) [HHSN275201800001I]
  5. Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD) [PJT-148621, PJT-159625]
  6. National Institute of Mental Health (NIMH)
  7. NIH
  8. NICHD
  9. Canadian Institutes of Health Research
  10. Mex-ican Government (Comisio ndeEquidadyGe nero de lasLegislaturas LX-LXI y Comisio ndeIgualdaddeGe nero de laLegislatura LXII de la H. Ca mara de Diputados de la Repu blicaMexicana)

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An inexpensive and easy-to-use rapid assay for detecting HLA-B*57:01 was developed in this study, which is of great significance for patients with HIV infection using ABC treatment.
The nucleoside reverse transcriptase inhibitor abacavir (ABC) is used commonly to treat young children with HIV infection and is a component of the fixed-dose-combination Triumeq(R). ABC can trigger a severe hypersensitivity reaction in people who are homozygous or heterozygous for HLA-B*57:01. Testing for HLA-B*57:01 before ABC initiation is standard-of-care in high-resource settings, but current tests are costly or difficult to access in resource-limited settings.To address these gaps, we developed an inexpensive simple-to-use rapid assay to detect HLA-B*57:01. We designed and optimized a multiplexed polymerase chain reaction (PCR) to amplify HLA-B*57 subtypes and the human beta-globin gene; employed probes and ligation to specifically tag the HLA-B*57:01 allele with biotin. Tagged-ligated products were detected by immunocapture in an enzyme-linked immunosorbent assay plate or lateral flow strip. Cell lines with known HLA genotypes were used to optimize the assay. The optimized assay was then compared with genotypes of clinical specimens (n = 60) determined by sequencing, with specimens enriched for individuals with HLA-B*57:01.The optimized assay utilizes 40-min 35-cycle multiplex PCR for B*57 and beta-globin; 20-min ligation reaction; and 15-min detection. Evaluation of the HLA-B*57:01 oligonucleotide ligation assay using clinical specimens had a sensitivity of 100% (n = 27/27 typed as B*57:01) and specificity of 100% (n = 33/33 typed as non-B*57:01) by visual interpretation of lateral flow strips. The cost is US$5.96/specimen. This rapid and economical assay accurately detects HLA-B*57:01 in clinical specimens. Use of this assay could expand access to HLA-B*57:01 genotyping and facilitate safe same-day initiation of ABC-based treatment.

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