Pham Tran Thu Ha, Tran Quang Binh, Sukasem Chonlaphat, Nguyen Van Dinh, Chu Chi Hieu, Do Thi Quynh Nga, Tran Ngoc Phuong Mai, Nguyen Hai Ha, Phung Thanh Huong
Hanoi University of Pharmacy, Hanoi, Vietnam.
National Institute of Nutrition, Hanoi, Vietnam.
Asian Pac J Allergy Immunol. 2024 Dec;42(4):375-381. doi: 10.12932/AP-201120-1000.
Severe cutaneous adverse drug reactions (SCARs) are rare but deadly drug reactions with severe damages to patients. One of the most well-known SCARs risk factors is the human leukocyte antigen (HLA) genes polymorphism. Among the HLA polymorphic alleles, the HLA-A33:03 allele has been found in association with SCARs induced by various drugs, especially in Asian people. There has not been any report on the specific detection protocol of the HLA-A33:03 allele.
This study aimed to design a nested AS-PCR protocol for detecting and distinguishing diplotype genotype of the HLA-A*33:03 allele.
A nested allele-specific (AS)-PCR protocol with four primer sets was designed. The method was compared with the Sanger sequencing method on 100 samples of unknown genotypes of unrelated Vietnamese people.
The nested AS-PCR method could identify the HLA-A33:03 allele and the HLA-A33:03 diplotype genotypes. Comparison with the Sanger sequencing method showed an absolute agreement (ĸ = 1.00, p < 0.001). The nested AS-PCR protocol had a sensitivity of 100% (95%CI: 92.13-100%) and a specificity of 100% (95%CI: 93.51-100%). The protocol was used for the determination of HLA-A33:03 allele distribution in 810 unrelated Vietnamese Kinh people, showing a frequency of HLA-A33:03 carriers of 19.6% and an allele frequency of 10.55%.
A novel nested AS-PCR method with a hundred-percent sensitivity and a specificity for the HLA-A*33:03 allele detection was reported. The protocol can be applied for the stratification of patients at SCAR risks with various drugs.
严重皮肤不良反应（SCARs）虽罕见但致命，会对患者造成严重损害。最著名的SCARs风险因素之一是人类白细胞抗原（HLA）基因多态性。在HLA多态性等位基因中，已发现HLA - A33:03等位基因与多种药物诱导的SCARs相关，尤其是在亚洲人群中。目前尚无关于HLA - A33:03等位基因特异性检测方案的报道。
本研究旨在设计一种巢式等位基因特异性PCR（AS - PCR）方案，用于检测和区分HLA - A*33:03等位基因的双倍型基因型。
设计了一种包含四组引物的巢式等位基因特异性（AS）- PCR方案。将该方法与桑格测序法在100例无关越南人未知基因型样本上进行比较。
巢式AS - PCR方法能够鉴定HLA - A33:03等位基因和HLA - A33:03双倍型基因型。与桑格测序法比较显示完全一致（ĸ = 1.00，p < 0.001）。巢式AS - PCR方案的灵敏度为100%（95%CI：92.13 - 100%），特异性为100%（95%CI：93.51 - 100%）。该方案用于810例无关越南京族人群中HLA - A33:03等位基因分布的测定，显示HLA - A33:03携带者频率为19.6%，等位基因频率为10.55%。
报道了一种对HLA - A*33:03等位基因检测具有100%灵敏度和特异性的新型巢式AS - PCR方法。该方案可用于对使用各种药物有SCAR风险的患者进行分层。
