应用套式PCR快速检测鸭坦布苏病毒.pdfVIP

上海市畜牧兽医学会 2011 年学术年会论文集 应用套式 PCR 快速检测鸭坦布苏病毒 颜丕熙，李国新，吴晓刚，闫丽萍，滕巧泱，李泽君 （中国农业科学院上海兽医研究所 农业部动物寄生虫学重点开放实验室，上海 200241） 摘 要：鸭坦布苏病毒是新发现的一种引起鸭产蛋下降、生长迟缓和死亡的黄病毒。目前，没有可靠的快 速诊断方法用于该病毒的检测。本研究根据鸭坦布苏病毒 E 基因序列，设计了两对重叠引物 P1、P2 和 P3、 P4，建立了检测鸭坦布苏病毒的套式PCR 方法。该套式 PCR 比一般的 PCR 敏感性高 10倍。采用该方法对安 徽省、河北省、山东省、浙江省、上海市等地发病鸭场的 63 份样品进行了检测，阳性检出率为 48/63（76.2% ）， 而病毒分离率仅为 13/63 （20.6% ）。上述研究表明与其他方法相比较本试验所建立的套式PCR 方法具有快 速、敏感、特异的优点，可用于鸭坦布苏病毒的流行病学调查及病毒的检测。 关键词：鸭坦布苏病毒；套式 PCR；鉴定 RAPID IDENTIFICATION OF DUCK TEMBUSU VIRUS BY THE NESTED POLYMERASE CHAIN REACTION YAN Pi-xi ，LI Guo-xin ，WU Xiao-gang，YAN Li-ping ，TENG Qiao-yang ，LI Ze-jun （Key Laboratory of Animal Parasitology, Ministry of Agriculture, Shanghai Veterinary Research Institute, CAAS, Shanghai 200241 ） Abstract: The duck Tembusu virus, a newly emerging Flavivirus, caused egg production decline, retarded growth, and even death among ducks in mainland China. However, there was no reliable, rapid and sensitive method to detect the virus. According to the sequence of the E gene, two pairs of primers P1, P2 and P3, P4 were designed, and a nested-PCR assay was established to detect the duck Tembusu virus. The results showed that the sensitivity of the nested-PCR was ten times higher than that of the general PCR. To test the reliability of nested-PCR assay, sixty-three clinical samples collected from ducks in Anhui, Hebei, Shangdong, Zhejiang, and Shanghai were tested. Forty eight samples were duck Tembusu virus positive by this method, while only thirteen of these samples were positive by Virus isolation. The result indicated that the nested-PCR assay was a sensitive, specific and rapid diagnosis method. Key words: Duck Tembusu virus; nested-PCR; identification 2010 年春季以来，上海、江苏、浙江、安徽等地相继爆发一种导致蛋鸭减产、生长迟缓 [1,2] 。该病 和死亡的一种新发病，研究证明引起该病的病原为坦布苏