马流感H3N8实时荧光RT-PCR检测方法的建立

Establishment of RT-PCR method for the detection of equine influenza virus H3N8

通过比对NCBI上马流感H3N8序列,针对HA和NA基因序列高度保守区域设计了特异性强的引物与探针,优化了程序,建立了以实时荧光RT-PCR技术检测马流感H3N8的方法。结果表明,该方法最小检测浓度为3.36×10^2copies/μL;特异性强,仅针对马流感H3N8样本检出为阳性;对58份临床样本的检测,阳性检出率为12.07%,是常规PCR的2.3倍,且与病毒分离100%符合。

By comparing the H3N8 sequence of Equine influenza virus (EIV) from NCBI,highly specific primers and probes were designed for the highly conserved region of HA and NA gene sequences,and the program was optimized to establish a RT-PCR method for the detection of EIV H3N8.The results showed that the minimum detection concentration was 3.36×10^2 copies/μL.With high specificity,only EIV H3N8 samples were positive.The positive detection rate of 58 clinical samples was 12.07%,2.3 times that of conventional PCR,and 100% consistent with virus isolation.