隐匿性HBV感染相关S基因突变对多种抗-HBs及HBsAg检测的影响

Effect of occult HBV infection related S gene mutation on detection of multiple anti-HBs and HBsAg

目的观察隐匿性HBV感染(OBI)S基因突变对HBsAg检测产生的影响。方法挑选9种典型的S基因突变株,从1例OBI患者及3例献血人员的血清进行分离。依次使用突变及野生型S基因重组质粒传染仓鼠的卵巢系统,深入分析入选的突变株HBsAg表达及突变蛋白对抗体和所用检测试剂反应性的影响。结果利用HBsAg试剂对不同突变株胞内HBsAg水平进行检测,与野生株相比,其水平显著降低。同一种样品采用抗-His抗体检测结果表明,与野生株对比,只有M1、M2、M5、M6、M7的HBsAg-His融合蛋白表达显著下降。与野生株比较,不同突变株与6种抗-HBs反应性显著下降,M1、M4、M7、M9和抗体4及M9出现的HBsAg和抗体6反应性水平最差。不同突变株和6种HBsAg试剂反应性与野生株相比显著降低,差异有统计学意义(P<0.05)。ELISA试剂内D、E、F漏检率依次为11.2%、22.3%、55.7%。结论突变HBsAg和抗-HBs结合力下降成为引起OBI表现的关键原因之一,通常使用HBsAg试剂对于突变HBsAg检测能力有一定不足之处,急需进行改进提升检测水平。

Objective To observe the effect of occult HBV infection(OBI)S gene mutation on HBsAg detection.Methods Nine typical S gene mutants were selected and isolated from the serum of one OBI patient and three blood donors.The ovarian system of the hamster was infected with the mutant and wild-type S gene recombinant plasmid in turn.The expression of the selected mutant HBsAg and the effect of the mutant protein on the reactivity of the antibody and the detection reagent were analyzed.Results The intracellular HBsAg level of different mutants was detected by HBsAg reagent.And the levels were significantly lower than those of wild-type strains.The test results of the same sample using anti-His antibody indicated that compared with that of the wild-type strain,only the expression of HBsAg-His fusion protein of M1,M2,M5,M6 and M7 was significantly decreased.Compared with that of the wild-type strains,the reactivity of different mutants with 6 anti-HBs was significantly decreased,and the reactivity levels of HBsAg in M1、M4、M7、M9 with antibody 4 and HBsAg in M9 with antibody 6 were the worst.The reactivity of different mutant strains with 6 HBsAg reagents was significantly lower than that of wild strains,and the difference was statistically significant(P<0.05).The missed detection rates of D,E,and F in the ELISA reagent were 11.2%,22.3%,and 55.7%,respectively.Conclusion The research shows that the decrease of the binding strength of mutant HBsAg and anti-HBs is one of the key reasons for the performance of OBI.Reagents generally used for the mutant HBsAg have certain shortcomings in detection capability.It is urgent to improve and improve the detection level.