Abstract Hepatitis B virus X (HBx) was a regulatory protein and it was coden by hepatitis B virus (HBV) genome. Serum HBx quantitative detection may be helpful for HBV, hepatocirrhosis, and hepatoma diagnosis. A HBx chemiluminesscence quantitative assay kit was produced following the recombinant HBx antigen expressed in E.Coli and the HBx monoclonal antibodies selected from HBx antigen immunizing mice. The sensibility, specificity and other characteristics of the HBx quantitative assay kit were analyzed. The purity of the recombinant HBx protein was ≥94%. Two of six monoclonal antibodies were selected and used for bait and detect antibody. The sensibility of the HBx quantitative assay kit is 0.1ng/ml, the linearity range of it is 0.5-600ng/ml. No coss-reactions to GLB, lipoprotein, HGB, HBc, HBe, HBs or HBV preS2 were observed. The addition accuracy was between 98.7%-105.7%, and the dilution accuracy was between 95.4%-104.5%. CV (within batch) ≤2.9, CV (batch to batch) ≤6.5. After keep in 37℃ for 4 days, the technical datas were almost same as keeping in 4℃. The HBx quantitative assay kit was use to detecte the HBx in 75 chronic HBV, 20 acute HBV, 25 hepatocirrhosis, 30 hepatoma patients’ serum and 180 healthy people’s serum. More than 55% chronic HBV patients, 68% hepatocirrhosis patients and more than 70% hepatoma patients can be detected. The higher grade of hepatocirrhosis patients classification it was, the higher concentration of the serum HBx was being. All the results show that the HBx chemiluminesscence assay kit we developed can not only be used for serum HBx quantitative detection, but also be used for the relationship between HBx and the hepatoma or the hepatocirrhosis study.
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