TRFIA、FQ-PCR方法联合检测乙肝标志物的应用探讨

Application of Combination of TRFIA and FQ- PCR in Detection of HBV Markers

目的探讨检测乙肝标志物联合使用时间分辨荧光免疫分析(TRFIA)和荧光定量聚合酶链反应(FQ-PCR)方法的应用价值.方法 2008年1月至2014年5月云南省第二人民医院检验科对门诊疑似乙肝患者2 476例,分别行TRFIA检测其血清学标志物HBV(HBV-M)和行FQ-PcR方法检测其HBv.DNA含量,并对2种方法的阳性检出率进行比较.结果 2 476例受检者中,FQ-PcR检测其HBv-DNA阳性1 656份,总阳性率为66.9%,其中TRFIA法HBsAg、HBeAg、HBcAb均阳性861份,使用FQ-PCR法检测HBV-DNA的含量阳性816份,占94.8%;TRFIA法HBsAg、HBeAb、HBcAb均阳性1 023例,使用FQ-PCR法检测HBV-DNA的含量阳性674份,占65.9%;HBsAg、HBeAg均阳性27份,其中使用FQ-PCR法检测HBV-DNA的含量阳性26份,占96.3%;HBsAg、HBcAb均阳性148份,其中使用FQ-PCR法检测HBV-DNA的含量阳性117份,占79.1%;HBsAb、HBeAb均阳性55份,其中使用FQ-PCR法检测HBV-DNA的含量阳性2份,占3.64%;HBsAb、HBeAb、HBcAb均阳性123份,其中使用FQ-PCR法检测HBV-DNA的含量阳性11份,占8.94%;HBeAb阳性22份,其中使用FQ-PCR法检测HBV-DNA的含量阳性2份,占9.09%;HBeAb、HBcAb均阳性43份,其中使用FQ-PCR法检测HBV-DNA的含量阳性3份,占6.98%;HBcAb阳性37份,其中使用FQ-PCR法检测HBV-DNA的含量阳性2份,占5.41%;HBsAb阳性89份,其中使用FQ-PCR法检测HBV-DNA的含量阳性2份,占2.22%;均阴性48份,其中使用FQ-PCR法检测HBV-DNA的含量阳性1份,占2.08%.结论 TRFIA法用来排查机体有无感染乙肝病毒,对于乙肝传染性强弱仅能做一个初步的估计指标,不能准确检测出血清HBV-DNA低滴度者,故靠TRFIA法诊断和判断病情是不够的;FQ-PCR法检查乙肝患者体内的乙肝病毒数量、复制指标,能够能为精确的判断出乙肝病毒在体内的复制、传染强弱情况,是对TRFIA法不足的补充.而FQ-PCR法在检测时也容易受到一些人为或非人为因素的影响.二者联合检测可提高其准确性,有利于病情的判断和诊治.

Objective To explore the application of combination of time-resolved fluorescence immunoassay（TRFIA） and fluorescence quantitative polymerase chain reaction（FQ- PCR） method in detection of HBV markers. Methods From January 2008 to May 2014 in Yunnan province,2476 cases of patients with suspected chronic hepatitis B in the Second People＇s Hospital of Yunnan Province were selected in this study. TRFIA was used to determine the HBV serological markers（HBV-m） and line FQ- PcR method was used to detect the HBV. DNA content,then the positive detection rates of the two methods were compared. Results In 2476 cases, positive HBv DNA detected with FQ- PcR was found in 1656, the total positive rate was 66.9%, positive HBsAg, HBeAg and HBcAb detected with TRFIA was found in 861, positive HBv DNA detected with FQ- PcR was found in 816,accounting for 94.8%;Positive HBsAg, HBeAb HBcAb detected with TRFIA was found in 1023 cases, positive HBv DNA detected with FQ- PcR was found in 674, accounting for 65.9%; Positive both HBsAg and HBeAg was found in 27,positive HBv DNA detected with FQ- PcR was found in 26（96.3%）. Positive HBsAg and HBcAb was found in 148,positive HBv DNA detected with FQ- PcR was found in 117, accounting for 79.1%. Positive both HBsAb and HBeAb was found in 55, positive HBv DNA detected with FQ- PcR was found in 2（3.64%）.HBsAb, HBeAb HBcAb were positive in 123, HBV- DNA was positive by FQ- PcR in 11, 8.94%; HBeAb wa positive 22,HBV- DNA was positive by FQ- PcR in 2, 9.09%; HBeAb HBcAb were positive in 43, HBV-DNA was positive by FQ- PcR in 3,accounting for 6.98%; HBcAb was positive in 37, HBV- DNA was positive by FQ- PcR in 2（5.41%）;HBsAb was positive in 89, HBV- DNA was positive by FQ- PcR in 2（2.22%）;All negative was found in 48,HBV- DNA was positive by FQ- PcR in 1,accounting for 2.08%. Conclusions TRFIA method can only give a preliminary estimation in screening hepatitis B virus（HBV） infection,cannot accurately detect the low serum HBV- DNA titer,so TRFIA is not enough for diagnosis of HBV infection. FQ-PCR method can accurately judge the replication of the hepatitis b virus in the body,infectious strength, is a complement to TRFIA method. While FQ- PPCR method in detection is easily affected by some artificial or non-artificial factors. Therefore,combining the two detection methods can improve the accuracy, is advantageous to diagnosis and judgment of HBV infection.