隐匿性乙型肝炎病毒感染检测的实验研究

Detecting the occult HBV inaction： A laboratory study

目的 了解乙型肝炎病毒表面抗原（HBsAg）阴性个体HBV DNA检出情况,探讨检测乙型肝炎病毒表面大蛋白（HBLP）用于筛查临床隐匿性HBV感染的血清学检测策略.方法 根据日常检测乙型肝炎病毒血清学模式（HBVM）结果,将HBsAg阴性标本分为乙型肝炎病毒表面抗体（HBsAb）阳性和阴性两大类,除外HBVM抗原抗体同时阳性等特殊模式,从临床检测备份管随机收集各1000份共计2000份血清标本,双份分装-20℃冻存;用多管混合的方法实施HBV DNA定量分析,筛选出阳性标本;HBV DNA阳性标本行HBLP检测,再经美国超敏MONOLISA HBsAg ULTRA试剂复检HBsAg.结果 1000份HBsAb阳性标本未检出HBV DNA阳性;1000份HBsAb阴性标本共检出19例HBV DNA阳性;19份HBV DNA阳性标本经HBLP检测和美国超敏试剂复检HBsAg均呈阳性.19份HBV DNA定量分析结果显示：大于500拷贝/ml 2例,400～500拷贝/ml 3例,300～400拷贝/ml 3例,200～300拷贝/ml 7例,100～200拷贝/ml 4例.结论 用国产ELISA试剂常规检测HBsAg漏检标本多来自HBsAb阴性个体;漏检个体HBLP结果可能阳性,检测HBLP有利于筛查隐匿性HBV感染;本研究为积极寻找临床隐匿性HBV感染的检测策略提供了血清学参考.

Objective To see the HBV DNA detection instance in the HBsAg negative people and to study the serological method detection strategy for detecting hepatitis B virus large surface protein （HBLP） to filtrate the occult HBV infection.Methods The HBsAg negative serum samples were divided into HBsAb negative and positive two species according to the hepatitis B virus markers （HBVM） in daily work excepting the special HBVM modes.Totsl 2000 stochastic serum samples with 1000 HBsAb negative results and 1000 HBsAb positive results were collected from the copy tubes to detect HBVM with national ELISA reagent kits and put them-20℃ frostily.Mixed samples（8 × 30μl）were analyzed with fluorescence quantitative PCR （FO-PCR）and filtrated the individual positive samples.The filtrated samples were doubly tested again with American MONOLISA HBsAg ULTRA reagents.Results No HBV DNA positive results were found out from the 1000 HBsAb positive samples and 19 cases HBV DNA positive results were found out from the 1000 HBsAb negative samples.On these 19 samples,the HBsAg results from the American MONOLISA HBsAg ULTRA reagents were all positive and the HBLP results were all positive,too.The 19 HBV DNA quantitative results were divided into 2 cases more than 500 copies/ml,3 cases between 400-500 copies/m1,3 cases between 300-400 copies/ml,7 cases between 200-300 copies/ml and 4 cases between 100-200 copies/ml.Conclusion The leaked samples tested HBsAg with national reagents are mostly from the HBsAb negative people.HBLP results may be positive on these samples and detecting HBLP marker is propitious to filtrate the occult HBV infection.This study provided a kind of serological reference for actively searching for the detecting strategy in occult HBV infection field.