摘要
目的 探讨丙型肝炎病毒（ＨＣＶ） 不同区抗原的反应性，为建立抗ＨＣＶ 酶免疫测定（ＥＩＡ） 确认试剂提供依据。方法 利用基因工程技术，重组所表达的ＨＣＶ 不同区抗原片段（ ＨＣＶＣ、ＮＳ３ 、ＮＳ４ 、ＮＳ５） ，建立了ＨＣＶ 单片段抗原ＥＩＡ 检测方法，检测了７４７ 份四川地区献血员血清。对其中３７３ 份血清，用２ 个厂生产的经批检合格的抗ＨＣＶＥＩＡ 试剂进行检测比较，对个别结果不符的样品进一步进行逆转录聚合酶链反应（ＲＴ－ ＰＣＲ） 分析。结果 献血员中抗ＨＣＶＣ 等４ 种抗体阳性检出率分别为４ ．６８ ％ （３５／７４７） 、５ ．２ ％ （３９／７４７） 、１ ．１ ％ （８／７４７） 、１ ．２ ％ （９／７４７） ，２ 家试剂阳性检出率均为４ ％（１５／３７３） 。结论 ＨＣＶ 不同区抗原片段的抗体检出率差异较大，ＮＳ３ 检出率最高，其次为Ｃ 区、ＮＳ５ 、ＮＳ４ 。说明抗ＨＣＶＥＩＡ 检测试剂以ＨＣＶ ＮＳ３ 区和Ｃ 区为主要抗原片段。
Objective To testify the reactivity of antigens from different regions of HCV as a basis for the development of a confirmation anti HCV EIA test kit.Methods We used recombinant HCV antigens from the Core, NS3, NS4, NS5 regions to formulate a set of anti HCV EIA test reagents. The reagents were used to screen 747 serum samples of donors from Sichuan province. Another 373 serum samples from donors that had been screened simultaneously by the Ministry of Health checked anti HCV EIA kits from two different companies. Samples that showed inconsistent results between the two kits were further tested using antigens from these four regions, and RT PCR was performed to check for the presence of HCV RNA.Results The percentage of sera positive for antibodies against various HCV antigens among samples from commercial blood donors was respectively against Core 4.68% (35/747), NS3 5.2% (39/747), NS4 1.1% (8/747), and NS5 1.2%(9/747). The positive detection rates for the EIA kits from the two companies were 4% (15/373).Conclusions Large differences were noted in anti HCV detection rates when using HCV antigens from different regions of the virus as capture reagents. That from NS3 giving the highest, then comes the Core, NS5 and NS4. In anti HCV EIA assay reagents, NS3 and Core are the most dominant and essential antigens.
基金
国家九五科技攻关项目
关键词
丙型肝炎
抗原
酶免疫试剂
HCV
Hepatitis C viruses Immunoenzyme techniques Antigens, viral
