摘要
根据GenBank中的猪圆环病毒2型ORF2基因序列,设计合成2对引物,通过对环介导等温(LAMP)扩增条件的优化,敏感性、特异性试验,成功建立了猪圆环病毒2型LAMP诊断方法。敏感性结果显示,建立的LAMP诊断方法最低核酸检测量为0.30 pg/L,而PCR诊断方法最低核酸检测量为30 pg/L;特异性结果显示,猪圆环病毒1型、猪细小病毒、猪伪狂犬病毒、猪繁殖与呼吸综合征病毒、猪瘟病毒的扩增结果均为阴性。对95份自然感染病猪样品的检测结果表明,该LAMP方法检测结果与PCR检测结果符合率为95.8%。整个扩增反应可在30 min内完成,结果肉眼可见,为猪圆环病毒2型的现场快速诊断提供了一种简便可行的方法,能够满足基层现场快速检疫的需要。
According to the gene sequences in GenBank of PCV-20RF2 gene, two pairs of specific primers were designed for amplifying PCV-2 ORF2 gene. By optimizing the reaction condition of LAMP and sensitivity test, specificity test, diagnosing method for PCV2 was established. The sensitivity test showed that the minimum amount of nucleic acid was 0.30 pg/L for LAMP, while 30 pg/L for PCR. Specificity test indicated that PCV-1, PPV, PRV, PRRSV and CFSV were not detected. The detection results of 95 samples from natural infection swine indicated that coincidence rate of LAMP method test results and PCR detection results was 95.8%. The amplification of LAMP could be accomplished within 30 min. And also the results could be detected by naked eyes. LAMP is a simple and rapid method for diagnosing PCV-2, and wwiu meet the needs of rapid quarantine for grassroots field.
出处
《广东农业科学》
CAS
CSCD
北大核心
2014年第11期163-166,共4页
Guangdong Agricultural Sciences
基金
贵阳市科技局现代农业与农村科技计划项目(筑科合同[2012102]5-1号)
贵州省科技厅农业攻关项目(黔科合NY字[2010]3085)
