HAV RT-PCR检测方法的建立及其在猪群中的流行调查

摘要

目的：了解屠宰生猪中甲型肝炎的流行情况，探讨猪作为HAV另一宿主和携带者的可能性，以期为人类甲肝的防治提供一定的理论依据。方法：采用酶联免疫吸附试验（ELISA）法对大理地区屠宰生猪血清中HAV IgG检测，逆转录-聚合酶链反应（RT-PCR）对屠宰生猪的全血和胆汁标本进行HAV RNA检测，以甲型肝炎患者的血液和胆汁为阳性对照标本。结果：①屠宰猪血清样本中检测出HAV的抗体阳性率为73.3%（374/510）；②建立了甲型肝炎病毒的RT-PCR检测方法；③用RT-PCR在阳性对照标本中检出HAV RNA；④用RT-PCR在屠宰猪的全血和胆汁样本中未检出HAV RNA。结论：成功建立起RT-PCR检测待测样本中的HAV RNA的方法；屠宰猪血清样本中检测出HAV的抗体阳性率较高；猪是否可以感染人的HAV还有待于进一步研究。%Objective:To study the epidemic state of the Hepatitis A Virus(HAV)infection in swinery, and to explore the possibility of swine as another host or carrier of HAV for providing basis for prevention and controlling measures of human HA. Methods:Serum HAV-IgG of swine in Dali was detected by enzyme-linked immunosorbent assay(ELISA), and HAV-RNA of swine blood and swine bile samples were tested by reverse transcription-polymerase chain reaction(RT-PCR), with the blood and bile of HAV as positive control. Results:①HAV antibody positive rate of the blood samples of swine were detected as 73.3%(374/510);②RT-PCR method was built to detect HAV; ③HAV-RNA of positive control samples were detected with RT-PCR; ④HAV-RNA of swine blood and swine bile samples were not detected with RT-PCR. Conclusion: RT-PCR method is successfully built to detect HAV-RNA. The antibody positive rate of HAV is rather high in the blood sample of swine. Further research is necessary to demonstrate whether the swine could infect human HAV.