巢式聚合酶链反应检测SEN病毒方法的建立及初步应用

Establishment and application of nested-polymerase chain reaction for detecting SEN virus

目的:研究建立巢式聚合酶链反应(nPCR)检测SEN病毒D和H亚型的方法,并将其应用到流行病调查。方法:选择SENV开放读码框1(ORFⅠ)核苷酸序列设计合成特异性引物,建立检测SEN病毒D和H型感染的巢式PCR方法,对216例无偿献血者、65例甲肝患者、152例乙肝患者、79例丙肝患者和83例非甲～非戊肝患者进行了SENV感染的检测。结果:该方法特异性和灵敏度均较高,检测结果显示健康献血者、甲肝、乙肝、丙肝和非甲～非戊型肝炎患者SENVD和/或H总感染率分别为29.1%、38.5%、55.3%、54.4%、38.5%。SENV D和/或H在健康献血者中总感染率显著低于乙肝和丙肝患者(P<0.01)。甲肝、乙肝、丙肝和非甲～非戊型肝炎患者总感染率无显著性差异(P>0.05)。结论:巢式PCR方法可用于检测SENV感染。我国部分地区无偿献血和肝炎等患者中存在SENV感染,其致病性有待进一步研究。

Objective：To establish a nested-polymerase chain reaction（nPCR） method for detecting SENV subtypes（SENV-D and SENV-H）.Methods：The specific primers were designed from highly conservative sequence of SENV D＆H,two pairs of which were used to establish a nested-PCR.Serum of 216 healthy blood donors and 65 patients with hepatitis A,152 with hepatitis B,79 with hepatitis C,and 83 with non A-E hepatitis were detected by this method.Results：The specificity and sensitivity of the nested-PCR test were perfect.The positive rates of SENV DNA in different populations were：29.1% in healthy blood donors,38.5% in patients with hepatitis A,55.3% in patients with hepatitis B,54.4% in patients with hepatitis C,38.5% in patients with hepatitis non A-E.SENV D/H infection was more frequent in patients with hepatitis B and hepatitis C than in healthy blood donors（P〈0.01）,but showed no significant difference between patients with hepatitis A,B,C and non A-E（P〉0.05）.Conclusion：This nested-PCR can be applied in the detection of SEN virus and the epidemiological survey of SENV-D/H in China.