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Performance verification of a time-of-flight mass spectrometry based cardiovascular drug-related polygene detection system

桑格测序 质谱法 基因型 多基因 人口 基因组DNA 色谱法 化学 医学 生物 DNA测序 遗传学 DNA 数量性状位点 基因 环境卫生
作者
Zhaohui Liu,Kai Cui,Linmei Yang
出处
期刊:Chinese Journal of Laboratory Medicine 卷期号:43 (1): 51-57
标识
DOI:10.3760/cma.j.issn.1009-8158.2020.01.005
摘要

Objective Toestablish and verify the method of genetic polymorphisms using time-of-flight mass spectrometry as a polygene testing platform. Methods 998 cases of DNA samples and 20 cases of whole blood samples were collected from Fuwai hospital of Chinese academy of medical sciencesduring September 2017 to October 2018, including 512 cases of males and 506 cases of females.280 patients aged 18-30, 442 patients aged 31-64, and 296 patients aged ≥65.11 cardiovascular drugsrelatedgenes in 998 DNA samples were detected by time-of-flight mass spectrometry to evaluate the compliance rate compared with identifiedresults. 20whole blood samples were selected to detect 11 genes using both time-of-flight mass spectrometry and Sanger sequencing. The results were compared twice, and accuracy was evaluated according to Sanger sequencing as the gold standard. Ten cases of genomic DNA with wild-type loci were selected for specific evaluation by time-of-flight mass spectrometry. Samples containing all heterozygous genotypes were measured after gradient dilution to evaluate the detection sensitivity of the new method. Samples containing all 49 genotypes (two genotypes were not found because they are rare in Chinese population) were used in order to do the inter-assay and intra-assay precision evaluation. An anti-interference study was performed by selecting wild and homozygous mutant samples of represented heterozygous peak shape. Results The results showed that the compliancerate of the single retrospective sample was over 99.5%. The resultsof time-of-flight mass spectrometry and Sanger sequencing was the same. The minimum detection limit of DNA was 0.4 ng, the inter-assay and intra-assay precision were 100%, and the degradation ability of the UNG enzyme was 105 copies/μl in aerosol.The reaction system has a strong anti-interference ability to the genome and intermediate aerosol, and no cross-contamination between different matrices of the chip. Conclusions The time-of-flight mass spectrometry as a polygene detection system shows agood detection performance and can be applied to clinical detection. In addition, this paper established a performance verification research scheme based on the time-of-flight mass spectrometry platform polygene detection system. Key words: Mass spectrometry; Genetic testing; Multilocus sequence typing; Cardiovascular agents
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