【摘要】 目的：建立乙型肝炎病毒（ｈｅｐａｔｉｔｉｓＢｖｉｒｕｓ，ＨＢＶ）ＰｒｅＳ抗原检测方法。方法：表达与纯化了ＨＢＶＰｒｅＳ１／２抗原，并用纯化蛋白免疫家兔，利用纯化抗血清包被ＥＬＩＳＡ板，吸附病人血清中包含ＰｒｅＳ１／２抗原的ＨＢＶ病毒颗粒或病毒亚单位，特异吸附颗粒用抗ＨＢｓＡｇ的酶标单抗检测。结果：利用金属螯合柱层析的方法高度纯化了ＰｒｅＳ重组抗原，用复性的重组抗原免疫家兔３次，血清抗体滴度可达１∶５１．２万。利用纯化抗血清建立的ＰｒｅＳ１／２抗原测定方法具有较高的灵敏度与特异性。在３００份ＨＢｅＡｇ阳性与１９２份ＨＢｅＡｇ阴性血清中分别检测到２６０份（８７％）与１４２份（７４％）阳性血清。结论：初步结果表明，乙型肝炎病人血清中ＰｒｅＳ１／２抗原的检测与ＨＢｅＡｇ的状况无关更多还原

【Abstract】 Objective: To set up hepatitis B virus PreS antigen assay. Methods: HBV PreS1/2 Antigen was over expressed and purified from E.coli. The purified protein was refolded and used to immunize rabbit. The rabbit anti sera were purified and coated to ELISA plates for specific adsorption of PreS1/2 containing HBV particles or subviral particles, which were subsequently detected by HRP conjugated anti HBs antibodies. Results: PreS1/2 antigen was highly purified by using Ni NTA agarose. The purified protein induced a titer of 1∶512 000 specific antibodies in rabbit after 3 times of immunization. The PreS1/2 antigen ELISA using the purified antibodies has been demonstrated to specifically detect PreS1/2 antigen in HBV patient sera. The PreS1/2 antigen was detected in 260 of 300 HBeAg positive sera (87%) and 142 of 192 HBeAg negative sera (74%). Conclusion: The presence of PreS1/2 antigen in HBsAg positive sera has no correlation with the status of HBeAg.更多还原