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A Novel Allele-Specific PCR Protocol for the Detection of the HLA-C*03:02 Allele, a Pharmacogenetic Marker, in Vietnamese Kinh People

Authors Pham TTH, Tran QB, Sukasem C, Nguyen VD, Chu CH, Do TQN, Tran NPM, Phung TH

Received 13 November 2020

Accepted for publication 18 January 2021

Published 9 February 2021 Volume 2021:14 Pages 27—35


Checked for plagiarism Yes

Review by Single anonymous peer review

Peer reviewer comments 2

Editor who approved publication: Prof. Dr. Martin H. Maurer

Tran Thu Ha Pham,1 Quang Binh Tran,2 Chonlaphat Sukasem,3– 5 Van Dinh Nguyen,6,7 Chi Hieu Chu,8 Thi Quynh Nga Do,9 Ngoc Phuong Mai Tran,9 Thanh Huong Phung1

1Hanoi University of Pharmacy, Hanoi, Vietnam; 2National Institute of Nutrition, Hanoi, Vietnam; 3Division of Pharmacogenomics and Personalized Medicine, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand; 4Laboratory for Pharmacogenomics, Somdech Phra Debaratana Medical Center (SDMC), Ramathibodi Hospital, Bangkok, Thailand; 5The Thai Severe Cutaneous Adverse Drug Reaction (THAI-SCAR) Research Group, Bangkok, Thailand; 6Respiratory, Allergy and Clinical Immunology Unit, Internal Medicine Department, Vinmec Times City International Hospital, Vinmec Healthcare System, Hanoi, Vietnam; 7College of Health Sciences, VinUniversity, Hanoi, Vietnam; 8Center of Allergology and Clinical Immunology, Bach Mai Hospital, Hanoi, Vietnam; 9National Institute of Hygiene and Epidemiology, Hanoi, Vietnam

Correspondence: Thanh Huong Phung; Quang Binh Tran Tel +84 904112588
; +84 904470844

Background: Allopurinol, a common anti-hyperuricemia drug, is well known as an inducer of severe cutaneous adverse drug reactions (SCARs). One of the most well-defined risk factors of allopurinol-induced SCARs is the presence of polymorphic alleles of human leukocyte antigen (HLA) genes, such as HLA-B*58:01 and HLA-C*03:02 alleles. There is no commercial test or published in-house protocol for the specific detection of the HLA-C*03:02 allele. In this article, we established for the first time a simple allele-specific (AS) PCR method to identify HLA-C*03:02 allele carriers, and at the same time, determine their zygosities.
Methods: A two-step AS-PCR protocol, using four primer sets, was designed to specifically amplify and differentiate the HLA-C*03:02 allele from 17 other HLA-C alleles found in Vietnamese people. The protocol was validated with PCR-sequencing-based typing (SBT) of 100 samples of unknown genotypes.
Results: The PCR protocol can detect the HLA-C*03:02 allele and determine the zygosity. The results of this protocol were highly consistent with those of the SBT (ĸ = 0.98, p < 0.001). Regarding the specific detection of the HLA-C*03:02 allele, the PCR protocol had a sensitivity of 100% (95% CI: 91.61– 100%) and specificity of 98.3% (95% CI: 90.9– 99.7%). The protocol was used to determine the distribution of the HLA-C*03:02 allele in 810 unrelated Vietnamese Kinh people, 14.2% of which were HLA-C*03:02 carriers, the allele frequency was 7.5%.
Conclusion: A novel AS-PCR protocol with a sensitivity of 100% for the detection of the HLA-C*03:02 allele was established. The protocol can be used for personalized treatment with allopurinol in order to minimize the risk of SCARs in Vietnamese people as well as in other Asian populations with similar genetic characteristics.

Keywords: severe cutaneous adverse reactions, HLA-C*03:02, allopurinol, HLA genotyping, AS-PCR

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