一种新型乙型肝炎病毒RNA定量检测方法的临床检测性能评估

Evaluation of clinical utility of a novel method for quantitative detection of hepatitis B virus RNA

目的评估一种新型乙型肝炎病毒(hepatitis B virus,HBV)RNA定量检测方法——HBV实时荧光核酸恒温扩增检测(simultaneous amplification and testing,SAT)(RNA捕获探针法)的临床检测性能。方法采用简单随机抽样法收集2017年6月至2018年6月上海复旦大学附属华山医院收治的170例慢性乙型肝炎患者和30例非HBV感染者的血清标本HBV RNA,使用HBV SAT与常规反转录聚合酶链反应(polymerase chain reaction,PCR)进行检测,对两种方法检测的灵敏度、特异度、к值及定量值相关性进行比较,并对两种方法检测的不同HBV DNA浓度样本的检出率进行分析比较。统计学分析采用χ^2检验。结果以临床诊断为标准,HBV SAT检测灵敏度为97.06%(165/170),特异度为100.00%(30/30),κ值为0.908;反转录PCR检测灵敏度为92.94%(158/170),特异度为100.00%(30/30),κ值为0.798。在HBV DNA低浓度样本(HBV DNA<100 IU/mL)中,HBV SAT检出率为77.27%(17/22),反转录PCR检出率为59.09%(13/22)。两种方法定量结果线性相关系数r=0.9878。结论全自动HBV SAT与反转录PCR定量结果一致,在HBV DNA低浓度样本中的检测灵敏度比反转录PCR方法略高,是一种快速、准确的HBV RNA定量检测方法。

Objective To evaluate the clinical utility of a novel quantitative assay named hepatitis B virus(HBV)simultaneous amplification and testing(SAT)using a kit for HBV RNA detection(RNA probes).Methods HBV RNA was detected in 170 serum samples of chronic hepatitis B patients and 30 serum samples of patients without HBV infection collected by simple random sampling method from June 2017 to June 2018 in Huashan Hospital,Fudan University,Shanghai using HBV SAT and reverse transcription polymerase chain reaction(PCR)method.The sensitivity,specificity,kappa value and quantitative correlation of the two methods were analyzed and compared.The detection rates of HBV RNA from samples with different HBV DNA concentrations of the two methods were analyzed and compared.Statistical analysis was performed by chi-square test.Results Based on the clinical diagnosis,the detection sensitivity,specificity,kappa value of HBV SAT were 97.06%(165/170),100.00%(30/30)and 0.908,respectively,while the reverse transcription PCR were 92.94%(158/170),100.00%(30/30)and 0.798,respectively.Among samples with lower concentration of HBV DNA(HBV DNA<100 IU/mL),the detection rates of HBV SAT and reverse transcription PCR were 77.27%(17/22)and 59.09%(13/22),respectively.The linear correlation coefficient of the two methods was r=0.9878.Conclusions Quantitation results of HBV RNA by HBV SAT and reverse transcription PCR method are consistent.HBV SAT is a rapid and accurate method for HBV RNA quantitative detection,which has a slightly higher detection rate than reverse transcription PCR in samples with low concentration of HBV DNA.