高灵敏乙型肝炎病毒核酸检测在术前筛查中的应用价值

The application value of high-sensitivity HBV nucleic acid test in preoperative screening

目的探讨高灵敏乙型肝炎病毒(HBV)核酸检测在术前筛查中的应用价值。方法收集术前筛查患者的血清标本,分别采用高灵敏荧光定量PCR法(高敏PCR)和化学发光微粒子免疫分析法(CMIA),检测血清中的HBV DNA和HBsAg,两方法结果不一致的标本采用Roche Cobas AmpliPrep/Cobas TaqMan全自动核酸检测系统检测HBV DNA。结果519例血清标本中,HBsAg阳性标本90例(17.3%),按HBsAg定量结果分为5组:I组(>5 000 IU/mL)、II组(1 000~5 000 IU/mL)、III组(500~1 000 IU/mL)、IV组(100~500 IU/mL)和V组(0.05~100 IU/mL),I组HBV DNA高于II、III、IV、V组,差异有统计学意义(P<0.05)。90例HBsAg阳性标本中有13例标本的HBV DNA低于高敏PCR法的最低检测限,占阳性标本的14.4%。HBV DNA阳性标本79例(15.2%),其中2例标本HBsAg为阴性,CMIA法的漏检率为2.5%。15例定性结果不一致的标本经Roche全自动核酸检测系统复核后,13例HBV DNA阴性而HBsAg阳性标本的HBV DNA均低于该方法的最低检测限,2例HBV DNA阳性而HBsAg阴性标本的HBV DNA高于该方法的最低检测限。与金标准相比,高敏PCR法检测HBV DNA的灵敏度、特异度、阳性预测值(PPV)和阴性预测值(NPV)均为100.0%;CMIA法检测HBsAg的灵敏度、特异度、PPV、NPV分别为97.5%、97.0%、85.6%、99.5%;高敏PCR法检测HBV的特异度和PPV高于CMIA法(P<0.05)。结论高敏荧光定量PCR检测HBV DNA可用于术前筛查,优于目前常规使用的CMIA法,能准确反映患者体内HBV的复制情况和传染性,及时发现隐匿性感染,有效防止医源性传播。

Objective To explore the application value of high?sensitivity Hepatitis B virus(HBV)nucleic acid detection in preoperative screening.Methods Serum samples were collected before surgery,and HBV DNA and HBsAg in serum were detected by high?sensitivity fluorescence quantitative PCR(high?sensitivity PCR)and chemiluminescent microparticle immunoassay(CMIA).Specimens with inconsistent results from both methods were tested for HBV DNA using the Roche Cobas AmpliPrep/Cobas TaqMan automated nucleic acid detection system.Results Among 519 serum samples,90 cases(17.3%)of HBsAg?positive specimens were divided into 5 groups according to the quantitative results of HBsAg:group I(>5 000 IU/mL),group II(1 000~5 000 IU/mL),group III(500~1 000 IU/mL),group IV(100~500 IU/mL)and groupⅤ(0.05~100 IU/mL).The viral load of HBV DNA in group I was higher than that in II,III,IV and V,and the difference was statistically significant(P<0.05).Among the HBsAg?positive specimens,there were 13 specimens with which the viral load of HBV DNA were lower than the minimum detection limit of the high?sensitivity PCR.79 specimens were positive for HBV DNA by high?sensitivity PCR,and the positive detection rate was 15.2%,of which 2 cases were negative for HBsAg,and the missed detection rate of CMIA method was 2.5%.15 cases with inconsistent qualitative results were reviewed by Roche??s automatic nucleic acid detection system,the HBV DNA of 13 specimens with HBV DNA negative but HBsAg positive were lower than the detection limit of the method.2 cases with HBsAg negative but HBV DNA positive were higher than the detection limit of the method.Compared with the gold standard,the sensitivity,specificity,positive predictive value(PPV)and negative predictive value(NPV)of high?sensitivity PCR were 100.0%,and the sensitivity,specificity,PPV and NPV of CMIA was 97.5%,97.0%,85.6%,and 99.5%respectively.The specificity and PPV of high?sensitivity PCR was higher than that of CMIA and the difference between them was statistically significant(P<0.05).Conclusions High?sensitivity fluorescence quantitative PCR detection of HBV DNA can be used for preoperative screening,which is superior to the conventionally used CMIA method,which can accurately reflect the replication and contagiousness of HBV in patients,detect occult infections in time,and effectively prevent iatrogenic transmission.