摘要
为建立猪传染性胃肠炎病毒(TGEV)NASBA的检测方法,本研究利用BioEdit和BlastN,根据GenBank中猪TGEV的S基因保守序列设计引物,建立了扩增TGEV的NASBA检测技术。利用该方法建立的反应体系在41℃反应2h即可得到有效扩增。实验结果表明:该方法对TGEV进行扩增获得约200bp特异性目的条带,而对CSFV、PRRSV、PCV-2、PPV、PEDV和ST细胞扩增结果均为阴性。NASBA的灵敏度高于普通RT-PCR,与病毒分离方法相当,可检测到5pg的核酸。该方法为TGEV的早期诊断提供了新途径。
A nucleic acid sequence-based assay （NASBA） for the detection of Porcine transmissible gastroenteritis virus （TGEV） was established using primers specific to TGEV S gene. The assay could specifically amplify a 198-base DNA fragment from the TGEV positive samples, while no product was generated from the phylogenetically or clinically related viruses and ST cell. The detecting limit of the assay was 5 pg nucleic acid, which is more sensitive than PCR method. In addition, the results of the NASBA was shown to correlate with those obtained by virus isolation.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2010年第6期451-454,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
国家质检总局科研项目(2009IK027)
重庆市自然基金项目(CSTC
2008BB1150)
关键词
猪传染性胃肠炎病毒
NASBA检测方法
Porcine transmissible gastroenteritis virus
nucleic acid sequence-based amplification diagnosis method
