逆向HBV DNA斑点杂交临床推广应用、方法改进及评价

摘要

The assay of HBV DNA reverse spot hybridization has been more simplified by using the filter with crude pBR322-HBV DNA spot,which was prepared by NaOH/SDS method.As a marker of HBV infection.the serum of hepatitis patients from several hospitals in Shanghai were assayed.Three hundred eighty-three samples of hepatitis patients with HBV immunoserological markers and HBV DNA spot assay were analysed by double blind test.It indicated that all the 63 cases of HBsAg(+)/HBoAg(+)/HBV DNA(q-)and 23 out of the 28 cases of HBsAg(q-)/HBoAg(-)/HBV DNA(+)showed positive reaction;28 cases of non-HBV hepatitis including 11 HAV and 6 NANB patients were negative.It is proved that this assay is a sensitive dependable marker for identifying the infectious status of HBV carriers.%本文成功地应用粗制pBR322-HBV DNA制备滤膜,进行逆向斑点杂交,从而进一步简化了方法,省却了CsCl超离心纯化重组质粒的繁复过程.为该方法商品化创造了条件.通过双盲法,收集临床肝炎血清进行逆向斑点杂交测定,对具有免疫血清标志及正向HBV DNA斑点杂交资料的383例血清的分析,发现69例:KBsAg(+)/HBeAg(+)/DNA正向斑试(+)血清100%显示阳性结果;28例HBsAg(+)/HBeAg(一)/DNA正向斑试(+)中有23例呈阳性反应;受检的28例各项HBV血清标志皆阴性的非乙型肝炎的肝病患者(其中11例为甲型肝炎,6例为非甲非乙型肝炎)逆向斑点杂交全部阴性,从而证明该方法确为灵敏、可靠的确定HBV感染性的有效手段,在乙型肝炎的诊断与防治中有重要的实用价值.