摘要
建立RT PCR检测猪瘟病毒的方法。根据已发表的猪瘟病毒E2基因 (囊膜糖蛋白gP55基因 )序列 ,设计合成了一对特异性引物 ,扩增片段的大小为 50 7bp ,用RT PCR技术对石门系标准株和 1 0株分离株进行检测。结果这对引物对标准株和 1 0株分离株均能扩增出与预期大小相符 50 7bpRT PCR产物 ,而对其他 6种猪病病原核酸的扩增结果为阴性。该RT PCR可检出 1 0 0pg的猪瘟病毒RNA模板 ,对人工感染猪不同组织样品进行检测 ,结果对白细胞抽提的核酸样品检出率最高为 1 0 0 % (2 4 / 2 4 ) ,其次为扁桃体、脾、肾 ,检出率为 83 3 % (2 0 / 2 4 ) ,再者为淋巴结 ,检出率为66 7% (1 6/ 2 4 )。对送检的 1 9份疑似猪瘟的病死猪病料组织进行RT PCR检测 ,结果有 1 6份样品为猪瘟病毒阳性。兔体交叉反应试验结果RT PCR阳性的 1 6份病料中 ,有 1 4份样品被判为含有猪瘟病毒 。
A reverse transcriptase polymerase chain reaction (RT PCR) method was developed for detection of hog cholera virus (HCV). A pair of specific primers were designed according to the sequence of E2 gene of HCV and a 507 bp long cDNA fragment of HCV was amplified. Using this pair of primers the RNAs of reference and 10 isolates strains were amplified, but other six swine pathogenic virus and bacterium were not amplified. As little as 100pg of HCV RNA was detectable by the RT PCR. In several tissues from infected swine, detection rate of HCV RNAs was highest in the leukocytes (100%), the next was tonsil, spleen, kidney (83 3%), and the lymph (66 7%). In the rabbit cross reaction test, there were 14 samples positive from the 16 samples which were amplified by the RT PCR
出处
《畜牧与兽医》
北大核心
2002年第11期11-14,共4页
Animal Husbandry & Veterinary Medicine
