一种联合检测乙型肝炎病毒前S1抗原与核心抗原方法的建立及其与病毒核酸检测结果的一致性

Establishment of a New Combined Enzyme Immunoassay for Detection of HBV PreS1 and Core Antigens and the Consistency with HBV DNA Test

本研究以与血清中HBV DNA含量高度相关的两种HBV抗原（前S1抗原与核心抗原）为靶标,建立了联合检测这两种HBV核酸相关抗原（NRAg）的双抗体夹心法ELISA试剂。对系列稀释血清的检测表明,该试剂的平均分析灵敏度为103.2基因组拷贝/mL（95%可信限102.2-4.2基因组拷贝/mL）,显著高于前S1抗原或核心抗原的单独检测。对994份HBsAg阴性血清的检测结果表明NRAg ELISA的特异性为99.7%（95%可信限：99.1%-99.9%）。对271份临床慢性肝炎血清进行检测,结果NRAg ELISA与HBV DNA结果的总符合率达96.3%（95%可信限：93.3%-98.2%）,NRAg ELISA的读值/临界值比（S/CO）与HBV基因组拷贝数呈正相关。利用NRAg试剂,发现了1例HBsAg“a”抗原表位突变的变异株。这些结果显示HBV NRAg ELISA与HBV DNA具有高度相关性,并能够检测出HBsAg抗原变异株,有望成为HBsAg变异株筛选的有力工具,并为广大基层医疗单位提供一种便捷的替代HBV DNA定性检测的手段。

In this study,a new combined enzyme immunoassay（NRAg ELISA） for detection of HBV PreS1 and core antigens which was highly consistent with serum HBV DNA test was established. The serial serum dilution test indicated that the average sensitivity of the assay was 10^3.2 genome copies/mL（95%CI： 10^2.2-4.2 genome copies/mL） ,which was notably higher than the test performed on Pre S1 or core antigen alone. The test with sera from 994 blood donors whose HBsAg were negative demonstrated that the specificity of this assay was 99.7% （95%CI： 99.1%-99.9%）. 271 serum samples from chronic hepatitis patients were also examined and the result showed that the total consistent rate between NRAg ELISA and HBV DNA was 96.3%（95%CI：93.3%-98. 2%）. The NRAg ELISA S/CO（signal/cutoff） was closely correlated with HBV genome copies（R=0. 9158,n=231）. Furthermore,by using this assay,we found a patient whose HBsAg was negative but HBV DNA was positive. Sequencing result showed that HBV genome from this patient had a point mutation in the ＂a＂epitope of S gene. Our results indicate that HBV NRAg ELISA has a high relativity with HBV DNA test,and can effectively detect the mutation of HBsAg,it is expected to be a potent tool for screening HBsAg mutant and is a convenient method for substituting HBV DNA test.