A Novel Allele-Specific PCR Protocol for the Detection of the HLA-C*03:02 Allele, a Pharmacogenetic Marker, in Vietnamese Kinh People
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 Pe...
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| Format: | Article |
| Language: | English |
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Dove Medical Press
2021-02-01
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| Series: | The Application of Clinical Genetics |
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| Online Access: | https://www.dovepress.com/a-novel-allele-specific-pcr-protocol-for-the-detection-of-the-hla-c030-peer-reviewed-article-TACG |
| _version_ | 1819162217181872128 |
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| author | Pham TTH Tran QB Sukasem C Nguyen VD Chu CH Do TQN Tran NPM Phung TH |
| author_facet | Pham TTH Tran QB Sukasem C Nguyen VD Chu CH Do TQN Tran NPM Phung TH |
| author_sort | Pham TTH |
| collection | DOAJ |
| description | 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, VietnamCorrespondence: Thanh Huong Phung; Quang Binh Tran Tel +84 904112588; +84 904470844Email huongpt@hup.edu.vn; tranquangbinh@dinhduong.org.vnBackground: 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 |
| first_indexed | 2024-12-22T17:24:43Z |
| format | Article |
| id | doaj.art-e1cee9f67c5244f39e3c4e55c4ba9e66 |
| institution | Directory Open Access Journal |
| issn | 1178-704X |
| language | English |
| last_indexed | 2024-12-22T17:24:43Z |
| publishDate | 2021-02-01 |
| publisher | Dove Medical Press |
| record_format | Article |
| series | The Application of Clinical Genetics |
| spelling | doaj.art-e1cee9f67c5244f39e3c4e55c4ba9e662022-12-21T18:18:45ZengDove Medical PressThe Application of Clinical Genetics1178-704X2021-02-01Volume 14273561858A Novel Allele-Specific PCR Protocol for the Detection of the HLA-C*03:02 Allele, a Pharmacogenetic Marker, in Vietnamese Kinh PeoplePham TTHTran QBSukasem CNguyen VDChu CHDo TQNTran NPMPhung THTran 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, VietnamCorrespondence: Thanh Huong Phung; Quang Binh Tran Tel +84 904112588; +84 904470844Email huongpt@hup.edu.vn; tranquangbinh@dinhduong.org.vnBackground: 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-PCRhttps://www.dovepress.com/a-novel-allele-specific-pcr-protocol-for-the-detection-of-the-hla-c030-peer-reviewed-article-TACGsevere cutaneous adverse reactionshla-c*03:02allopurinolhla genotypingas-pcr |
| spellingShingle | Pham TTH Tran QB Sukasem C Nguyen VD Chu CH Do TQN Tran NPM Phung TH A Novel Allele-Specific PCR Protocol for the Detection of the HLA-C*03:02 Allele, a Pharmacogenetic Marker, in Vietnamese Kinh People The Application of Clinical Genetics severe cutaneous adverse reactions hla-c*03:02 allopurinol hla genotyping as-pcr |
| title | A Novel Allele-Specific PCR Protocol for the Detection of the HLA-C*03:02 Allele, a Pharmacogenetic Marker, in Vietnamese Kinh People |
| title_full | A Novel Allele-Specific PCR Protocol for the Detection of the HLA-C*03:02 Allele, a Pharmacogenetic Marker, in Vietnamese Kinh People |
| title_fullStr | A Novel Allele-Specific PCR Protocol for the Detection of the HLA-C*03:02 Allele, a Pharmacogenetic Marker, in Vietnamese Kinh People |
| title_full_unstemmed | A Novel Allele-Specific PCR Protocol for the Detection of the HLA-C*03:02 Allele, a Pharmacogenetic Marker, in Vietnamese Kinh People |
| title_short | A Novel Allele-Specific PCR Protocol for the Detection of the HLA-C*03:02 Allele, a Pharmacogenetic Marker, in Vietnamese Kinh People |
| title_sort | novel allele specific pcr protocol for the detection of the hla c 03 02 allele a pharmacogenetic marker in vietnamese kinh people |
| topic | severe cutaneous adverse reactions hla-c*03:02 allopurinol hla genotyping as-pcr |
| url | https://www.dovepress.com/a-novel-allele-specific-pcr-protocol-for-the-detection-of-the-hla-c030-peer-reviewed-article-TACG |
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