Rapid detection of CYP2C9, CYP2C19,CYP4F2,VKORC1 and ABCB1 gene polymorphisms by liquid phase chip technology
10.3760/cma.j.issn.1009-9158.2019.12.013
- VernacularTitle: 液相芯片技术快速检测CYP2C9、CYP2C19、CYP4F2、VKORC1及ABCB1基因多态性
- Author:
Hongli XU
1
,
2
;
Rentang DENG
1
;
Meilian CHEN
1
;
Zaixin CHEN
1
;
Zhihong HUANG
1
;
Bo SITU
3
;
Guixing KONG
1
;
Lisha LAI
1
;
Lei ZHENG
3
;
Wenjin FU
1
Author Information
1. Department of Laboratory, Houjie Hospital, Dongguan 523945, China
2. Department of Laboratory, Hunan Wugang People, s Hospital, Shaoyang 422400, China
3. Department of Laboratory Medicine Centre, Southern Medical University, Guangzhou 510515, China
- Publication Type:Journal Article
- Keywords:
Liquid phase chip technology;
Allele-specific primer extension;
Warfarin;
Clopidogrel;
Single nucleotide polymorphism
- From:
Chinese Journal of Laboratory Medicine
2019;42(12):1042-1050
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a method for simultaneous and rapid detecting of the polymorphisms in Cytochrome P450 2C9 (CYP2C9), CYP2C19, CYP4F2, Vitamin K epoxide reductase (VKORC1) and ATP-binding cassette subfamily B member1 (ABCB1) gene, which were associated with warfarin and clopidogrel, based on liquid phase chip technology.
Methods:Method establishment. The eight gene sequences near targeted sites related to warfarin and clopidogrel were found in Genbank, and the specific primers and probes were designed. Through multiple PCR amplification, followed by allele specific primer extension (ASPE), and MagPlex-Tag microspheres hybridization, the suspension array Luminex 200 system step-by-step, the genotypes were determined by fluorescence signal. The reaction system was optimized and its methodological evaluation was performed. 260 patients with antithrombotic therapy from Dongguan houjie hospital were recruited in this study form June 2017 to December 2018. The eight genotypes of the 260 patients were detected by the established method, and the results were compared with the sequencing results.
Results:The results of 260 samples showed that allelic median fluorescence intensity (MFI) ratios of homozygotes (mutant/wild-type) were all greater than 0.9 or less than 0.1, and all the allelic MFI ratios of heterozygotes were between 0.3 and 0.6. The within run and between run coefficients of variance for allelic MFI ratios were lower than 6.4% and 10.9%, respectively. The minimum DNA template requirements was 0.75ng. The genotypes of 260 patients determined by the established method were completely concordant with the sequencing results.
Conclusion:A method was established successfully for rapid detecting the genotypes which associated with warfarin and clopidogrel based on liquid phase chip technology.
