实时荧光核酸恒温扩增试验和定量反转录-聚合酶链反应对血清乙型肝炎病毒RNA定量检测的一致性评价  

Evaluation of simultaneous amplification testing and quantitative reverse transcription-polymerase chain reaction for quantitative detecting serum hepatitis B virus RNA

作　　者：黄晨璐 许伟[1] 胡乾坤 张小楠[1] 李强[1] 黄玉仙[1,2] 陈良 HUANG Chenlu;XU Wei;HU Qiankun;ZHANG Xiaonan;LI Qiang;HUANG Yuxian;CHEN Liang(Shanghai Public Health Clinical Center,Fudan University,Shanghai 201508,China;Department of Infectious Diseases,Huashan Hospital,Fudan University,Shanghai 200040,China)

机构地区：[1]复旦大学附属公共卫生临床中心,上海201508 [2]复旦大学附属华山医院感染科,上海200040

出　　处：《微生物与感染》2020年第3期158-165,共8页Journal of Microbes and Infections

基　　金：上海市科委医学引导类(中、西医)科技支撑项目(17411969700);上海申康医院发展中心市级医院新兴前沿技术联合攻关项目(SHDC12015129)。

摘　　要：为评价实时荧光核酸恒温扩增试验(simultaneous amplification testing,SAT)与定量反转录-聚合酶链反应(quantitative reverse transcription-polymerase chain reaction,qRT-PCR)检测血清样本中乙型肝炎病毒(hepatitis B virus,HBV)RNA的相关性与一致性,本研究收集了在复旦大学附属公共卫生临床中心就诊的212例患者的血清样本,其中慢性乙型肝炎(chronic hepatitis B,CHB)患者HBV DNA≥100 IU/mL 81例、HBV DNA<100 IU/mL 76例,以及作为对照的非HBV感染患者55例。采用SAT和qRT-PCR方法分别检测所有血清样本中的HBV RNA,对检测结果进行统计学分析。在HBV DNA≥100 IU/mL的CHB患者中,SAT和qRT-PCR的HBV RNA阳性检出率均为95.06%(77/81)。2种方法具有较好的相关性与一致性(R 2=0.803和CCC=0.882)。Bland Altman分析显示绝对偏倚为0.1 log copies/mL,相对偏倚为0.97%。配对t检验显示,结果无统计学差异(t=1.617,P=0.110)。在HBV DNA<100 IU/mL的CHB患者血清样本中,HBV RNA阳性检出率不同,SAT为98.68%(75/76),qRT-PCR为88.16%(67/76),2种方法相关性与一致性较差(R 2=0.326和CCC=0.438)。Bland Altman分析显示绝对偏倚为0.5 log copies/mL,相对偏倚为0.86%。配对t检验显示,结果有统计学差异(t=3.654,P<0.001)。在非HBV感染对照患者中,SAT和qRT-PCR均未检出HBV RNA阳性。结果提示,在HBV DNA≥100 IU/mL的CHB患者中应用SAT与qRT-PCR检测血清HBV RNA,其相关性与一致性较好,在HBV DNA<100 IU/mL的CHB患者中相关性与一致性存在一定差异。The purpose of the current study is to evaluate the correlation and consistency of simultaneous amplification testing(SAT)and quantitative reverse transcription-polymerase chain reaction(qRT-PCR)for quantitative detecting serum hepatitis B virus RNA.212 serum samples from Shanghai Public Health Clinical Center including 81 CHB patients with HBV DNA≥100 IU/mL,76 CHB patients with HBV DNA<100 IU/mL and 55 patients without HBV infection were detected by SAT and qRT-PCR methods.In HBV DNA≥100 IU/mL CHB patients,95.06%(77/81)samples were positive both in SAT and qRT-PCR method.SAT showed a relevantly good correlation and comparability with qRT-PCR(R 2=0.803,CCC=0.882).Bland Altman analysis shows absolute bias was 0.1 log10 copies/mL and relative bias was 0.97%.The paired T test analysis of test results had no difference(t=1.617,P=0.110).In HBV DNA<100 IU/mL CHB patients,98.68%(75/76)samples were positive in SAT method and 88.16%(67/76)samples were positive in qRT-PCR method.The statistical analysis of test results had difference(P<0.001).SAT showed a relevantly bad correlation and comparability with qRT-PCR(R 2=0.326,CCC=0.438).Bland Altman analysis shows absolute bias was 0.5 log10 copies/mL and relative bias was 86%.The paired T test analysis of test results had difference(t=3.654,P<0.001).In 55 patients without HBV infection,all samples were negative in SAT method and qRT-PCR method.Correlation analysis shows strong correlation and consistency between SAT and qRT-PCR for CHB patients with HBV DNA≥100 IU/mL.Difference was found in the measurement of RNA using SAT and qRT-PCR for CHB patients with HBV DNA<100 IU/mL.

关 键 词：乙型肝炎病毒 核糖核酸 定量反转录-聚合酶链反应 实时荧光核酸恒温扩增试验 

分 类 号：R373.2[医药卫生—病原生物学]