摘要
本研究根据GenBank中登录的猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)M基因保守序列6个特异性部位设计了2对引物,建立了PRRSV的环介导逆转录等温检测方法。结果表明,该方法灵敏度比RT-PCR高100倍;全部反应可在1 h内完成并可得到其特有的阶梯状条带,而且猪圆环病毒、猪瘟病毒、猪伪狂犬病毒的扩增结果均为阴性;在反应体系中添加SYBR GREENⅠ染料后,可通过肉眼观察有无荧光而直接判定结果。该方法灵敏度高、特异性好,可作为猪繁殖与呼吸综合征病毒的快速诊断方法。
A series of specific primers targeting the 6 distinct sequences of M gene were designed based on the sequences of porcine reproductive and respiratory syndrome virus（PRRSV） GP6（M） gene available in GenBank.A reverse transcriptase loop-mediated isothermal amplification（RT-LAMP） for the rapid detection of PRRSV was developed by using the specific primers.The results showed that the detective sensitivity of RT-LAMP method was higher than that of the two-step RT-PCR.The amplification could be accomplished within 1 h with good specificity and high sensitivity,while no product was generated with porcine circovirus,classic swine fever virus and pseudorabies virus.With the addition of SYBR GREENⅠ,the PT-LAMP of PRRSV could be detected by naked eyes.
出处
《中国畜牧兽医》
CAS
北大核心
2009年第12期53-56,共4页
China Animal Husbandry & Veterinary Medicine
基金
教育部"创新团队计划"(2007-68)
番禺科技局(2008-z-60-1)资助
关键词
猪繁殖与呼吸综合征病毒
M基因
环介导逆转录等温扩增
porcine reproductive and respiratory syndrome virus
M gene
reverse transcriptase loop-mediated isothermal amplification（RT-LAMP）
