摘要
为了进行大鲵蛙病毒(Chinese giant salamander ranavirus,CGSRV)的早期诊断,以US22家族基因作为靶基因,建立大鲵蛙病毒Taq Man MGB实时荧光定量PCR检测方法,同时对该方法进行特异性、敏感性、重复性评估,并对73份未知大鲵血液样品进行检测。结果显示,标准曲线在所选浓度范围内具有良好的线性关系,相关系数为0.997;该方法对大鲵蛙病毒的检测有高度特异性,与其他8种病毒或细菌之间均无交叉反应;灵敏性良好,能检测到2.00×101copies/L的标准品浓度,比普通PCR高1 000倍;批内和批间重复的变异系数均小于2%,重复性好;在73份未知样品检测中,Taq Man MGB实时荧光定量PCR能够检测到更低浓度的病毒,较常规PCR的敏感性更佳。鉴于该方法快速、特异、敏感、可重复、可定量的优点,对大鲵蛙病毒早期潜伏感染的快速诊断具有重要意义。
A sensitive and specific Taq Man MGB real-time PCR assay for detection on US22 family gene of Chinese giant salamander ranavirus（CGSRV）was established.The specificity, sensitivity and repeatability of this technology were assessed.And 73 unknown salamander blood samples were detected using this method. The results showed that the standard curve had an excellent linear correlation（with a R2 value of 0.997）.The established method was highly specific for amplification of CGSRV, without cross-reactions with other 8 kinds of viruses or bacteria. A minimum of 2.00×101copies/ L of standards could be detected, which of the sensitivity of real-time PCR is about 1 000 times higher than that of the conventional PCR test.The coefficients of variance（CV） were less than 2%, which indicated a decent reliability. In the detection on 73 unknown salamander blood samples, Taq Man MGB quantitative PCR could detect a lower concentration of positive infection than that of the traditional PCR method.The Taq Man MGB real-time PCR allows for a rapid, specific, sensitive, repeatable, and quantitative detection, and will be useful for early detection of CGSRV.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2016年第6期749-755,共7页
Chinese Veterinary Science
基金
四川省科技支撑资助项目(2014NZ0027)
