新型乙型肝炎病毒基因分型检测试剂盒的临床评价

The evaluation of one new hepatitis B virus genotypes detecting kit

目的评价一种新型乙型肝炎病毒基因分型检测(免疫荧光法)试剂的临床性能。方法收集中国人民解放军三○二医院研究对象的血清383例,与血清对应的血浆50例。乙型肝炎病毒A基因型评价采用北京万泰生物药业股份有限公司试剂盒(荧光免疫层析法),基因序列测定为对照方法;乙型肝炎病毒B、C、D基因型评价使用北京万泰生物药业股份有限公司试剂盒(荧光免疫层析法),上海之江生物科技试剂盒(荧光定量PCR法)为对照试剂。验证试剂和对照试剂同时检测同一例标本,若结果不一致,2种方法分别进行复测,结果以复测为准,如仍不一致进行基因测序,以测序结果为准,对结果进行Kappa一致性检验,计算2种方法的检测一致率。另通过验证试剂检测对应的血浆标本,评价验证试剂对血清、血浆标本检测能力的一致性。结果 A基因型验证试剂检出15例,对照方法检出12例,2种方法的符合率为12/15。B基因型验证试剂相对对照试剂的阳性符合率为98.00%(49/50),阴性符合率为99.06%(315/318),总符合率为98.91%(364/368),Kappa值为0.954。C基因型验证试剂相对对照试剂的阳性符合率为98.78%(242/245),阴性符合率为86.99%(107/123),总符合率为94.84%(349/368),Kappa值为0.881。D基因型验证试剂相对对照试剂的阳性符合率为87.50%(7/8),阴性符合率为100.00%(360/360),总符合率为99.73%(367/368),Kappa值为0.932。验证试剂对50例对应的血清/血浆标本检测,结果均为5例乙型肝炎病毒B基因型,44例C基因型,1例未分型。结论该新型试剂盒对乙型肝炎病毒的A、B、C、D型分型效果较好,可用于感染病毒分型的临床检测。

Objective To evaluate a new reagent called reverse dot blot assay kit for hepatitis B virus genotypes（immunofluorescence method）.Methods Three hundred and eighty-three serum samples were collected.Another 50 cases of corresponding plasma were needed.Hepatitis B virus genotype A to D was detected by the diagnostic kit for hepatitis B virus genotypes（fluorescence immunochromatography assay）.The gene sequencing technique was regarded as a control method for genotype A,and other genotypes were determined by real-time quantitive PCR.The same sample was detected by using the experimental reagent and control reagent at the same time.If the results were inconsistent,another two previous detections were needed as the final results.If the results of one more time detecting were still inconsistent,then the samples would be sequenced and the results were analyzed by kappa-test for symmetric measures.In addition,the results of the corresponding serum and plasma samples were analyzed to check the consistency.Results Fifteen genotype A samples were detected out by diagnostic kit,and 12 samples were detected out by the control reagent.For genotype B,C,D,the positive coincidence rates were of 98.00%,98.78%,87.50%,respectively;the negative coincidence rates were 99.06%,86.99%,100.00%,respectively;the total coincidence rates were 94.84%,94.84%,99.73%,respectively;the value of kappa were 0.881,0.881,0.932,respectively.And the detecting results of plasm samples were same as that of the corresponding serum samples.Conclusion The experimental reagent was able to meet the requirements to the clinical diagnosis.