A Novel Allele-Specific PCR Protocol for the Detection of the HLA-C03:02 Allele, a Pharmacogenetic Marker, in Vietnamese Kinh People

• Published in: Application of clinical genetics Vol. 14; pp. 27 - 35
• Main Authors: Pham, Tran Thu Ha, Tran, Quang Binh, Sukasem, Chonlaphat, Nguyen, Van Dinh, Chu, Chi Hieu, Do, Thi Quynh Nga, Tran, Ngoc Phuong Mai, Phung, Thanh Huong
• Format: Journal Article
• Language: English
• Published: New Zealand Dove Medical Press Limited 01-01-2021; Taylor & Francis Ltd; Dove; Dove Medical Press
• Subjects: Alleles; Allopurinol; as-pcr; Binding sites; Complications and side effects; Deoxyribonucleic acid; DNA; Ethylenediaminetetraacetic acid; Gene frequency; Genotypes; Histocompatibility antigen HLA; Histocompatibility antigens; hla genotyping; HLA histocompatibility antigens; hla-c03:02; Hyperuricemia; Methods; Original Research; Population genetics; Precision medicine; Risk factors; severe cutaneous adverse reactions; Tissue typing; Zygosity; Thailand; Vietnam; United States > US; HLA genotyping; HLA-C03:02; severe cutaneous adverse reactions; allopurinol; AS-PCR
• ISSN: 1178-704X; 1178-704X
• DOI: 10.2147/TACG.S278652

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 (
) genes, such as
and
alleles. There is no commercial test or published in-house protocol for the specific detection of the
allele. In this article, we established for the first time a simple allele-specific (AS) PCR method to identify
allele carriers, and at the same time, determine their zygosities.
A two-step AS-PCR protocol, using four primer sets, was designed to specifically amplify and differentiate the
allele from 17 other
alleles found in Vietnamese people. The protocol was validated with PCR-sequencing-based typing (SBT) of 100 samples of unknown genotypes.
The PCR protocol can detect the
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
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
allele in 810 unrelated Vietnamese Kinh people, 14.2% of which were
carriers, the allele frequency was 7.5%.
A novel AS-PCR protocol with a sensitivity of 100% for the detection of the
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.