用多聚酶链反应检测非洲猪瘟病毒的研究  

Determination of African Swine Fever Virus by polymerase chain reaction ( PCR )

作　　者：张兹钧 金由辛[2] 陈士友[3] 

机构地区：[1]拱北动植物检疫局,珠海519020 [2]上海生物化学研究所 [3]南京农业大学

出　　处：《中国兽医科技》1992年第12期3-6,共4页Chinese Journal of Veterinary Science and Technology

摘　　要：用HindⅢ对非洲猪瘟病毒核酸的重组质粒pRPEL2进行酶切后,将第5片段插入到pT7/T3α-19上而得到亚克隆重组质粒pT7 ASFH5。用双脱氧末端终止法测定了该插入片段的核苷酸序列。在此基础上选择了一段含194bp的片段作为扩增模板,并人工合成了该片段两端含20个碱基的寡核苷酸作为引物,从而建立了诊断非洲猪瘟病毒的多聚酶链反应。与核酸杂交相比,该方法不但更为快速和简便,而且灵敏度提高了一千万倍左右.After the recombination plasmid pRPEL2 for African Swine Fever ( ASF ) virus nucleic acid was cut using Hindi and then the fifth fragment was inserted into pT7/T3α-19, the subcl-one reco-plasmid pT7ASFH5 was obtained. The nucleotide sequence of the inserted fragment was measured with the double deoxidizing terminal cessation method. The PCR for diagnosing ASF was established using an artificial oligonucleotide fragment at which two ends are 20 bases as a primer and using a fragment containing 194 bp as amplified template. In comparison with nucleic acid hybridization, PCR was more rapid and simpler, and its sensitivity was raised 107 times.

关 键 词：猪瘟病毒 聚合酶链反应 诊断 猪病 

分 类 号：S858.282.6[农业科学—临床兽医学]