以PCR Ct值作为核酸扩增检测质量控制指标的可行性研究  

Feasibility study of PCR Ct value as quality control index for nucleic acid amplification test

作　　者：韩春花 朱琳 殷立奎 曹连连 丁国良 张超 延义芹 Han Chunhua;Zhu Lin;Yin Likui;Cao Lianlian;Ding Guoliang;Zhang Chao;Yan Yiqin(Dongying Centeral Blood Station,Dongying 257091,Shangdong Province,China;Dongying People′s Hospital,Dongying 257091,Shangdong Province,China)

机构地区：[1]东营市中心血站,东营257091 [2]东营市人民医院,东营257091

出　　处：《国际输血及血液学杂志》2023年第3期273-276,F0003,共5页International Journal of Blood Transfusion and Hematology

摘　　要：目的探讨以PCR循环数阈值(Ct)值作为核酸扩增检测(NAT)质量控制指标的可行性。方法选择2021年2至3月东营市中心血站用于NAT的223份乙型肝炎病毒(HBV)DNA(n=67,原始浓度为50 IU/mL),丙型肝炎病毒(HCV)RNA(n=78,原始浓度为200 IU/mL),人类免疫缺陷病毒(HIV)RNA(n=78,原始浓度为1000 IU/mL)质控品为研究对象。将HBV DNA、HCV RNA和HIV RNA质控品分别稀释6倍后,随当日血液筛查标本一同进行连续NAT,记录质控品Ct值,观察质控品Ct值的分布,并绘制Levey-Jennings质控图。采用不同浓度的质控品随当日血液筛查标本一同进行NAT,观察其浓度对数值与Ct值的线性关系;并根据线性方程确定质控品的最佳质控品浓度范围。对NAT所得Ct值采用Kolmogorov-Smirnov法进行正态性检测。结果①本研究HBV DNA、HCV RNA、HIV RNA质控品的Ct值分别为33.67±0.56、36.87±0.56和35.36±0.43,并且均符合正态分布(D=0.766、0.916、0.846,P=0.600、0.371、0.471)。②HBV DNA、HCV RNA、HIV RNA质控品Ct值的Levey-Jennings质控图结果显示,所有质控品的Ct值均符合Westgard多规则质控法。③HBV DNA质控品为50 IU/mL时,HBV DNA浓度(C)与Ct值的线性关系公式为logC=-0.229Ct+8.57,HBV DNA质控品的最佳浓度为10~1000 IU/mL。结论血站NAT工作中,合适浓度质控品的Ct值符合正态分布,可用于绘制质控图。并且可根据质控品浓度与其Ct值的线性关系公式,选择质控品的最佳测定浓度。ObjectiveTo explore feasibility of using PCR cycle threshold(Ct)value as quality control index of nucleic acid amplification test(NAT).MethodsFrom February to March 2021,a total of 223 quality control products with known concentration used for NAT at Dongying Central Blood Station were selected as research objects,including hepatitis B virus(HBV)DNA(n=67,original concentration was 50 IU/mL),hepatitis C virus(HCV)RNA(n=78,original concentration was 200 IU/mL),human immunodeficiency virus(HIV)RNA(n=78,original concentration was 1000 IU/mL).After the quality control products of HBV DNA,HCV RNA and HIV RNA were diluted sixfold respectively,NAT was carried out continuously together with the blood screening samples of the same day,Ct values of quality control products were recorded,the distribution of Ct values of quality control products was observed,and the Levey-Jennings quality control chart was drawn.Quality control products of different concentrations were used for NAT together with the blood screening samples of the same day,and the linear relationship between the concentration and Ct value of quality control products was calculated.And according to the linear equation to determine the best quality control product concentration range.The Ct values obtained from NAT were tested for normality using the Kolmogorov-Smirnov test.Results①In this study,the Ct values of HBV DNA,HCV RNA and HIV RNA were 33.67±0.56,36.87±0.56 and 35.36±0.43,respectively,and all of them accorded with the normal distribution(D=0.766,0.916,0.846,Z=0.766,P=0.600,0.371,0.471).②Levey-Jennings quality control chart of Ct values of HBV DNA,HCV RNA and HIV RNA quality control products showed that Ct values of all quality control products were in line with Westgard multi-rule quality control method.③When HBV DNA quality control product was 50 IU/mL,the linear relationship between HBV DNA concentration(C)and Ct value is logC=-0.229Ct+8.57,and the optimal concentration of HBV DNA quality control product was 10-1000 IU/mL.ConclusionsCt values

关 键 词：聚合酶链反应 质量控制 正态分布 核酸扩增检测 循环数阈值 

分 类 号：R440[医药卫生—诊断学]