【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。更多还原

【Abstract】 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 detection of RHDV in China.更多还原