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〔目的〕掌握南方口岸蚊媒携带病毒的本底资料,为蚊传疾病的预防控制工作提供依据。〔方法〕采用电动吸蚊器人工法和捕蚊磁场自动法采集南方5省口岸各类蚊虫。采集到的蚊类超低温送至实验室,研磨处理后用荧光PCR方法检测登革病毒、乙脑病毒、黄热病毒、西尼罗病毒、基孔肯雅病毒等重要蚊媒病毒,结果阳性的标本进一步进行PCR扩增和核苷酸序列测定分析;蚊标本研磨液同时用C6/36细胞进行虫媒病毒分离培养,出现细胞病变后分别用黄病毒科、甲病毒科各自的通用引物进行鉴定;对未能鉴定的未知病毒进一步用随机PCR方法进行扩增、克隆、序列测定、Blast搜索。〔结果〕从南方5省口岸采集到各类蚊虫12575只,鉴定后共分成254组。各组标本经荧光PCR方法检测,结果登革病毒、黄热病毒、西尼罗病毒、基孔肯雅病毒均为阴性;检测到2份福建省来源三带喙库蚊的标本乙脑病毒核酸阳性,经乙脑病毒E基因引物PCR扩增、测序分析证实为GⅠ型病毒。254份标本经C6/36细胞分离培养出现42份细胞病变,用黄病毒科、甲病毒通用引物PCR扩增,均未得到特异片段。选取1份典型病变的细胞培养物进行随机PCR鉴定,结果发现了1种潜伏于C6/36细胞中的浓核病毒。〔结论〕南方5省口岸蚊媒中可能未携带登革病毒、黄热病毒、西尼罗病毒、基孔肯雅病毒等重要蚊媒病毒,只有少量蚊虫携带乙脑病毒,蚊虫体内检测到的GⅠ型乙脑病毒属于福建省首次发现,出现病变的C6/36细胞可能是由自身潜伏的1种C6/36细胞浓核病毒引起。
[Objective] To master the background information of mosquito-borne virus in the southern port and provide basis for the prevention and control of mosquito-borne diseases. [Methods] Artificial mosquito electric mosquito and mosquito magnetic field automatic collection of all kinds of mosquitoes at the five southern ports. The collected cryopreserved mosquitoes were sent to the laboratory for testing. After grinding, fluorescent microscopy was used to detect important mosquito vectors such as Dengue virus, JE virus, Yellow fever virus, West Nile virus and Chikungunya virus. The result was positive The specimens were further subjected to PCR amplification and nucleotide sequence analysis. The mosquito specimens were also cultured in C6 / 36 cells for Arbovirus isolation and identification by the universal primers of Flaviviridae and Alphaviridae respectively after cytopathic effect ; Unknown unidentified viruses were further amplified by random PCR, cloning, sequencing, Blast search. [Results] 12 575 mosquitoes of all kinds were collected from the 5 southern ports of the country and were divided into 254 groups after identification. The samples of each group were detected by fluorescent PCR method. Results Dengue virus, yellow fever virus, West Nile virus and Chikungunya virus were negative. Two samples of Culex tritaeniorhynchus from Fujian Province were detected. Positive, by JE virus E gene primers PCR amplification, sequencing analysis confirmed that the G Ⅰ virus. A total of 42 cytopathic lesions were isolated and cultured in C6 / 36 cells from 254 samples, and no specific fragments were obtained by PCR using flaviviridae and alphavirus universal primers. A typical lesion cell culture was selected for random PCR identification. As a result, a single dendritic cell virus latent in C6 / 36 cells was found. CONCLUSION: The mosquito vectors in the five southern ports may not carry important mosquitoes such as dengue virus, yellow fever virus, West Nile virus and chikungunya virus, while only a few mosquitoes carry JE virus. In mosquitoes detected in mosquitoes GⅠhepatitis B virus belongs to Fujian Province for the first time found that C6 / 36 cells with pathological changes may be caused by their own latent concentration of C6 / 36cells.