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Background <p>The gold standard for detecting HLA-B*57:01 allele linked to Abacavir hypersensitivity is sequence-based typing (SBT), which is labour-intensive, not readily available and needs special expertise. HCP5 gene polymorphism is a surrogate marker for HLA-B*57:01 allele. Therefore, we compared, nested PCR for HLA-B*57:01 allele, HCP5 PCR with SBT method to develop a simple and cost-effective method for detection of HLA-B*57:01 allele.</p> Method <p>Whole blood cells were collected from PLHIV and Buffy coat and DNA was isolated. Direct PCR for HLA-B*57 gene followed by nested PCR for the HLA-B* 57:01 allele and HCP5 gene PCR was done in all patients. The PCR results of both methods were confirmed by the SBT method.</p> Result <p>Among 366 samples, 25 (6.83%) were positive for HLA-B* 57:01 allele by nested PCR. HCP5 gene PCR was positive in 25 (6.83%) individuals who were also positive for the HLA-B*57:01 allele with nested PCR. A strong correlation was observed between the HLA-B*57:01 allele and HCP5 with negative and positive predictive values of 100% respectively in our population.</p> Conclusion <p>Nested (two-step) and HCP5 gene PCR (single-step) are simple, rapid and cost-effective methods for HLA-B*57:01 allele detection and either can be adapted in the ART centre located in resource-poor settings.</p>

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Comparison of different methods for the detection of HLA-B* 57:01 allele in people living with HIV in Eastern Uttar Pradesh, India

  • 期刊论文

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  • 发表日期：

    2025年11月27日

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  • Tulika Kumari Rai,
  • Jaya Chakravarty,
  • Kumari Priyanka,
  • Abhilasha Gautam,
  • Shweta Srivastva

摘要

Background

The gold standard for detecting HLA-B*57:01 allele linked to Abacavir hypersensitivity is sequence-based typing (SBT), which is labour-intensive, not readily available and needs special expertise. HCP5 gene polymorphism is a surrogate marker for HLA-B*57:01 allele. Therefore, we compared, nested PCR for HLA-B*57:01 allele, HCP5 PCR with SBT method to develop a simple and cost-effective method for detection of HLA-B*57:01 allele.

Method

Whole blood cells were collected from PLHIV and Buffy coat and DNA was isolated. Direct PCR for HLA-B*57 gene followed by nested PCR for the HLA-B* 57:01 allele and HCP5 gene PCR was done in all patients. The PCR results of both methods were confirmed by the SBT method.

Result

Among 366 samples, 25 (6.83%) were positive for HLA-B* 57:01 allele by nested PCR. HCP5 gene PCR was positive in 25 (6.83%) individuals who were also positive for the HLA-B*57:01 allele with nested PCR. A strong correlation was observed between the HLA-B*57:01 allele and HCP5 with negative and positive predictive values of 100% respectively in our population.

Conclusion

Nested (two-step) and HCP5 gene PCR (single-step) are simple, rapid and cost-effective methods for HLA-B*57:01 allele detection and either can be adapted in the ART centre located in resource-poor settings.