Liu Qu, Lin Lin, Zhou Jian-Ming, Chen Ying-Jian, Zhang Qi-Wen, Wang De-Quan, Li Jing-Mei, Jin Yu-Juan
Longgang Center for Disease Control and Prevention of Shenzhen, Shenzhen 518172, China.
Bing Du Xue Bao. 2013 Jan;29(1):1-6.
During 2009-2012, the Nam Dinh virus (NDiv) was detected from the samples of Culex pipiens quinquefasciatus in Shenzhen China. In this study, cell culture,SYBR Green I based real time RT-PCR and RT-PCR were performed to analyze the cell susceptibility and other biological characteristics of the NDiV isolates. The results showed that C6/36 cell line was susceptible to four isolates of Culex pipiens quinquefasciatus. The "S" type amplification curve and specific melting curve were obtained in the realtime fluorescence quantitative RT-PCR based on SYBR Green I for the detection of the NDiV from the mosquito. The target bands from the RdRp gene and partial fragment of ZmHel1 gene were observed using agarose gel electrophoresis. Both the nucleotide and amino acid sequences of four Shenzhen isolates showed more than 99.00% homology with the Vietnam representative NDiV strain (02VN178). Phylogenetic analysis showed that four Shenzhen isolates shared the same evolution branch as the Vietnam representative NDiV strain. This is the first report of NDiV in China.
2009年至2012年期间,在中国深圳的致倦库蚊样本中检测到了南定病毒(NDiv)。在本研究中,进行了细胞培养、基于SYBR Green I的实时逆转录聚合酶链反应(RT-PCR)和逆转录聚合酶链反应(RT-PCR),以分析NDiv分离株的细胞易感性和其他生物学特性。结果表明,C6/36细胞系对4株致倦库蚊分离株敏感。基于SYBR Green I的实时荧光定量RT-PCR检测蚊体内的NDiv时,获得了“S”型扩增曲线和特异性熔解曲线。使用琼脂糖凝胶电泳观察到来自RdRp基因和ZmHel1基因部分片段的目标条带。深圳4株分离株的核苷酸和氨基酸序列与越南代表性NDiv株(02VN178)的同源性均超过99.00%。系统发育分析表明,深圳4株分离株与越南代表性NDiv株共享同一个进化分支。这是NDiv在中国的首次报道。