献血员血清抗－HCV－IgM检测及其意义

Detection of serum IgM antibody to hepatitis C virus and its significance in donors

目的探讨献血员血清抗－ＨＣＶ－ＩｇＭ与ＨＣＶ感染的关联性。方法献血员血清８３６份。以ＨＣＶ混合抗原包被聚苯乙烯反应板微孔，以辣根过氧化物酶标记的羊抗人ＩｇＭ单克隆抗体为酶标抗体，建立ＥＬＩＳＡ检测血清抗－ＨＣＶ－ＩｇＭ。以第二代ＥＬＩＳＡ法检测抗－ＨＣＶ－ＩｇＧ，以ＰＣＲ检测血清抗－ＨＣＶ－ＩｇＧ或抗－ＨＣＶ－ＩｇＭ阳性样品中的ＨＣＶ－ＲＮＡ。结果２－巯基乙醇破坏试验及ＨＣＶ抗原抑制试验结果显示，以所建立的ＥＬＩＡＳ检测到的为ＨＣＶ特异ＩｇＭ，批内变异系数为３．４７％，批间变异系数为７．０６％。８３６份献血员血清中，有２份抗－ＨＣＶ－ＩｇＧ阴性而抗－ＨＣＶ－ＩｇＭ和ＨＣＶ－ＲＮＡ均阳性。结论本研究中建立的ＥＬＩＳＡ可较好地用于检测血清抗－ＨＣＶ－ＩｇＭ、献血员筛选及防止丙型肝炎传播。

Objective To research the relationship between the serum IgM antibody to hepatitis C virus (IgM Anti-HCV) and HCV infection.Method A sensitive and specific ELISA was developed for detection of serum IgM Anti-HCV. The micro-titer plates were coated with a mixture of both the core and the non-structural peptides of HCV, and horseradish peroxidase fabled monoclonal antibody against human IgM was used. Serum samples from 836 donors were detected for IgM anti-HCV with this ELISA and for IgG anti-HCV with 2nd generation ELISA kits. HCV-RNA were detected with PCR in 抗-HCV IgG or IgM positive samples.Results Specificity of IgM anti-HCV detected with this ELISA was revealed by inhibition tests and 2-mercaptoethanol breach tests.CV of within -run and between-run was 3.47% and 7.06%,respectively.Out of 836 serum samples, 2 samples were positive for IgM anti-HCV and for HCV-RNA but negative for IgG anti-HCV.Conclusion The ELISA has better specificity and sensitivity for determination of serum IgM anti-HCV, and for screening donors with HCV infection to prevent hepatitis C from spreading.