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为建立快速检测2型鲤疱疹病毒(CyHV-2)的方法,本研究针对CyHV-2三联体蛋白基因的两段保守序列设计特异性引物,建立了CyHV-2双重PCR检测方法,并对反应条件进行优化,扩增的目的片段分别为218 bp和369 bp。特异性试验显示仅CyHV-2扩增出特异性片段,其它病毒均呈阴性;灵敏性试验表明该方法最低可以检测到3×102拷贝/μL的病毒量。对2012年~2013年采集自江苏、湖北、江西、宁夏等地的鲫鱼(Carassius auratus)样品进行检测,结果表明,该方法检测样品的阳性率为87%。以上结果表明建立的双重PCR方法具有良好的特异性和敏感性,可以用于CyHV-2的快速检测和病原流行病学调查。
In order to establish a rapid detection method for Cyclophilus herpesvirus type 2 (CyHV-2), a specific primer was designed according to the two conserved sequences of the CyHV-2 triad protein gene. A double PCR detection method of CyHV-2 was established. Conditions were optimized, the purpose of amplification fragments were 218 bp and 369 bp. Specific tests showed that only CyHV-2 amplified specific fragments, other viruses were negative; sensitivity test showed that the method can detect a minimum of 3 × 102 copies / μL of the virus. The Carassius auratus samples collected from Jiangsu, Hubei, Jiangxi, Ningxia and other places from 2012 to 2013 were tested. The results showed that the positive rate of this method was 87%. The above results show that the established dual PCR method has good specificity and sensitivity and can be used for rapid detection of CyHV-2 and epidemiological investigation of pathogens.