PCR检测猪输血传播病毒Ⅰ型方法的建立及其初步应用

Development and application of PCR for detection of swine Torque Teno virus genogroupⅠ

参照GenBank已公布的猪源输血传播病毒Ⅰ型(Torque Teno Virus genogroupⅠ,TTVⅠ)全基因序列,设计并合成一对特异性引物,PCR扩增片段为194bp,测序结果与已公布的猪TTVⅠ型序列同源性为92.6%。用所建立的方法检测猪圆环病毒Ⅱ型、猪细小病毒、猪伪狂犬病病毒、猪多杀性巴氏杆菌、猪链球菌等病原,无特异性条带出现,不存在交叉反应。该法灵敏度高,能检测到11.3pg的阳性核酸。检测某规模化猪场送检的44份健康母猪血清,结果TTVⅠ型的阳性率为13.6%(6/44);检测2006年7月到2007年3月期间采自广东、江西、湖南、广西、福建等5个省154份疑似"猪高热病"病料组织,结果TTVⅠ型的阳性率为40.9%(63/154),这表明我国规模化猪场TTVⅠ型感染率是非常高的。试验结果证明所建立的PCR方法为猪TTVⅠ的检测和流行病学调查提供有力的技术支持。

A pair of primers amplified swine Torque Teno virus genogroupⅠ（TTVⅠ） was designed and synthesized.The polymerrase chain reaction（PCR） was developed for detecting swine TTVⅠafter selecting the best reaction conditions.The PCR product obtained was about 194 bp and shared 92.6% identity with the published sequence.Negative results were obtained from PCVⅡ,PPV,PRV,PM and S.S.The 11.3 pg of swine TTVⅠ DNA was amplified by the sensitivity test.The PCR was applied to detect 44 healthy sow serum samples from a pig farm,TTVⅠ was detected in 13.6% samples（6/44）.154 clinical samples,which were suspected swine high fever syndrome from 5 provinces,were analysed by the method,the result showed that 40.9% samples（63/154） were swine TTVⅠ-positive.These results suggest that swine TTVⅠ was widespread in south china.This study supplied a technique for swine TTVⅠ detection and epidemiology research.