摘要
根据GenBank中的中国兔出血症病毒(RHDV)分离株VP60基因保守序列设计合成了内、外2对引物,在Bst DNA聚合酶的作用下完成等温核酸扩增反应,对反应条件逐一优化,并对临床样品进行检测。结果表明,该方法简便、快速、特异性好,灵敏性较常规反转录聚合酶链式反应(RT-PCR)高100倍。该方法可同时扩增7株RHDV中国分离株,说明其对检测国内RHDV分离株的有效性。对15份临床样品的检测结果显示,环介导等温扩增(LAMP)阳性检出率达80%,RT-PCR检测阳性率为60%。该LAMP方法具有灵敏度高、特异性强、操作简便快速、不需要复杂仪器设备、便于肉眼观察等优点,可作为临床检测RHDV的新方法,适合国内基层和实验室对RHDV的快速检测,具有广泛的推广应用价值。
Based on conserved sequence of rabbit hemorrhagic disease virus（RHDV） VP60 gene, two pairs of primers were designed and synthesized. After optimization, a loop-mediated isothermal amplification（LAMP） method was established and used to test clinical samples. The results indicated that this method was simple, rapid, performing at constant temperature, high specific, and 100-fold more sensitive than standard reverse transcription polymerase chain reaction（RT-PCR）. Specific amplification for 7 isolates of RHDV in China indicated that the LAMP was reliable. Clinical detection result of 15 samples showed 80% positive rate of LAMP and 60% positive rate of RT-PCR. The LAMP assay was a sensitive, specific and easy method with no need to the specialized equipment, which could provide a useful technique for clinic de- tection of RHDV in China.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2013年第4期390-395,共6页
Chinese Veterinary Science
基金
黑龙江省产业技术创新服务平台项目(PC10S04)
关键词
兔出血症病毒
环介导等温扩增
病毒检测
rabbit hemorrhagic disease virus
loop-mediated isothermal amplification
virus detection
