中文摘要：

目的建立检测患者血清中HBV的x抗原（HBxAg）及其抗体（anti-HBx）的方法，阐明其在慢性肝炎、肝硬化和肝癌患者血清中的临床意义。方法将HBV的x基因全长克隆至原核表达载体pET30a（＋）中，构建pET30a-X原核表达载体，转化人大肠杆菌BL21（DE3）后，经异丙基-β-D-硫代半乳糖苷（IPTG）诱导表达His-HBx融合蛋白，纯化后将重组蛋白免疫家兔，制备兔抗HBx多克隆抗体。应用纯化后的兔抗HBx多克隆抗体和纯化后的重组蛋白，建立检测血清中HBxAg和anti-HBx的方法。应用建立的ELISA法，分别对慢性乙型肝炎、肝硬化和肝癌患者组及正常对照组血清中HBxAg和anti-HBx进行检测。结果HBxAg／anti-HBx在慢性乙型肝炎、肝硬化和肝癌患者中的阳性率分别为8．7％／10．4％，17．9％／40．6％，9．8％／34．4％。统计学分析显示，与慢性乙型肝炎相比，肝硬化或肝癌患者血清中anti—HBx的阳性率较高（P〈0．01）。结合临床检测的其他HBV血清学指标，发现在anti-HBs阳性病例中，anti-HBx阳性率为5．2％（1／19）；而在HBsAg、HBeAg和anti-HBc3项同时为阳性的病例中，anti-HBx的阳性率为28．9％（39／135），二者相比，差异具有统计学意义（P〈0．05）。213名正常对照血清中HBxAg和anti-HBx均为阴性。结论血清中anti-HBx并非一个保护性抗体，标志HBV的感染和复制，是从慢性乙型肝炎向肝硬化和（或）肝癌发展的血清标志之一。

英文摘要：

Objective To establish a method of detecting hepatitis B virus x antigen （HBxAg） and antibody to HBxAg （anti-HBx） and to demonstrate its clinical significance of HBxAg and anti-HBx in sera from patients of chronic hepatitis B （ CHB）, liver cirrhosis （ LC ） and hepatocellular carcinoma （HCC）. Methods Full length HBx gene was cloned into pET30a （ ＋ ）, a prokaryotic expression vector, named pET30a-X. It was transformed into Escherichia coli BL21 （ DE3 ） , followed the fusion protein of HBx-His was induced by IPTG. The purified fusion protein was used to immunize rabbit as an antigen to generate polyclonal antibody to HBx protein. The method of enzyme-linked immunosorbent assay （ELISA） was established by using purified fusion protein and generated antibody, which was used to detect HBxAg and anti-HBx in sera from patients of CHB, LC, HCC and normal healthy people. Results The positive rates of HBxAg/anti-HBx were 8.7%/10.4% for CHB, 17.9%/40.6% for LC, and 9.8%/34.4% for HCC, respectively. In statistics, the positive rates of anti-HBx in LC and HCC were higher than that in CHB （P 〈 0. 01 ）. With the data from clinical examination of HBV markers in sera, it was found that the positive rate of anti-HBx was 5.2% （ 1/19 ） in anti-HBs ＋ cases, however, the positive rate of anti-HBx was 28.9% （39/ 135） in HBsAg ＋/HBeAg ＋/anti-HBc ＋ cases. The two positive rates were significant difference （P 〈 0.05 ）. Both HBxAg and anti-HBx were totally negative in the sera from normal healthy people （n =213） as negative control. Conclusions Anti-HBx in sera is not a protective antibody, showing infective and replication of HBV. The positive of anti-HBx in sera demonstrates the development from CHB to LC and HCC.