重庆地区动物携带Nipah病毒和Hendra病毒的检测

Epidemiological surveillance of Henipavirus in Chongqing

目的本研究拟建立一步法实时RT—PCR检测Nipah病毒（Nipah virus，NiV）和Hendra病毒（Hendra virus，HeV）的方法，对采集自中国重庆地区动物外周血样本进行NiV和HeV检测，以获得我国重庆地区的NiV和HeV的病毒流行病学资料。方法2007年6月起至2008年6月，采集自重庆地区饲养的猪外周血标本580份、奶牛外周血标本250份、山羊外周血标本180份，密度梯度离心法分离外周血淋巴细胞，Trizol法提取细胞总RNA。用已建立的NiV和HeV一步法实时RT—PCR检测方法，对RNA样本进行NiV和HeV检测。对阳性样本进行PCR产物序列鉴定及序列分析等研究，在可能的情况下进行病毒分离培养，并提交流行病学调查报告。结果对采集自中国重庆地区饲养的3种动物的样本进行NiV和HeV核酸检测，成功进行了一步法实时RT—PCR反应，所有样本荧光扩增曲线均无Takeoff点，曲线平坦，判为阴性结果。结论初步流行病学研究提示我国重庆地区饲养的动物猪、牛、羊中NiV和HeV未发现阳性。

Objective To establish nucleic acid testing techniques for detecting Nipah virus （NiV） and Hendra virus （HeV）, and to test the NiV and HeV in peripheral blood collected from domestic pigs, cows and goats in Chongqing. Methods Peripheral blood samples of 580 domestic pigs, 250 cows, 180 goats were collected from Chongqing since June 2007 to June 2008. The lymphocytes were separated by density gradient centrifugation and total RNA was extracted using Trizol method for detection of NiV and HeV with one-step real-time RT-PCR. Sequence identification and analysis were performed for positive PCR products. Virus isolation and culture were adopted for positive samples, and epidemiologic reports were submitted. Results Nucleic acid detections searching for NiV and HeV were successfully performed in animal blood samples collected from Chongqing. ＂Takeoff points＂ were not found in fluorescence amplification curves of all samples. Curves kept the same slope, and assays were judged as negative. Conclusion Until now, Neither NiV or HeV infection has been found in domestic animals blood samples collected from Chongqing, which suggest a lower possibility of outbreaks of Nipah disease and Hendra disease in Chongqing in the near future.