Tram Duong Bich, Chuong Ho Quoc, Phuong Huynh Anh, Phung Nguyen The Nguyen, Nguyen Mai-Lan, Vu Hoang Anh
Center for Molecular Biomedicine, University of Medicine and Pharmacy at Ho Chi Minh City, Ho Chi Minh City, Vietnam.
Pediatric Department, Faculty of Medicine, University of Medicine and Pharmacy at Ho Chi Minh City, Ho Chi Minh City, Vietnam.
J Lab Physicians. 2023 Jun 13;15(4):567-572. doi: 10.1055/s-0043-1768948. eCollection 2023 Dec.
The variants impact thiopurine dose selection in acute lymphoblastic leukemia patients. The ability to rapidly detect variants is important in clinical practice. This study aims to develop a simple polymerase chain reaction (PCR) procedure for detecting variants in Vietnamese patients. Sanger sequencing was used to determine variants from 200 patients. We designed primers and optimized the PCR procedure for detection of wild-type and variant alleles and compared with Sanger sequencing results. The inserted variant c.55_56insGAGTCG was detected by differences in size through conventional PCR. The tetra-primer amplification refractory mutation system PCR was successful in detecting two variations, c.52G > A and c.415C > T. The sensitivity and specificity of PCR procedure achieved 100% when compared to 200 Sanger sequencing results. Our PCR procedure is suitable for replacing Sanger sequencing to detect the variants in clinical setting.
这些变体影响急性淋巴细胞白血病患者硫嘌呤剂量的选择。在临床实践中，快速检测变体的能力很重要。本研究旨在开发一种简单的聚合酶链反应（PCR）程序，用于检测越南患者的变体。
采用桑格测序法确定200例患者的变体。我们设计了引物并优化了用于检测野生型和变异等位基因的PCR程序，并与桑格测序结果进行比较。
通过常规PCR检测到插入变体c.55_56insGAGTCG的大小差异。四引物扩增阻滞突变系统PCR成功检测到两个变异，即c.52G＞A和c.415C＞T。与200份桑格测序结果相比，PCR程序的敏感性和特异性均达到100%。
我们的PCR程序适用于替代桑格测序法在临床环境中检测变体。
