白纹伊蚊对登革病毒垂直传递能力研究

目的利用细胞及分子生物学技术，研究白纹伊蚊贵州省贵阳、兴义、毕节株对1—4型登革病毒（DEN1—4）的垂直传递能力。方法采集贵州省贵阳、兴义、毕节市市郊自然界白纹伊蚊，驯化3代。分别取羽化后3～5d龄成蚊为实验组，白纹伊蚊海南株为对照组；用DEN1—4国际参考株分别感染各组蚊虫，将感染后亲代及子1～3代成蚊制备蚊悬液，接种C6／36进行病毒分离并鉴定；间接免疫荧光法检测DEN抗原；常规方法提取总RNA。利用DENNS1基因区通用引物经逆转录-聚合酶链反应（RT—PCR）检测DEN核酸，限制性内切酶酶切分型。结果白纹伊蚊贵阳、兴义、毕节、海南株亲代蚊虫均对DEN易感；白纹伊蚊兴义株与海南株均能垂直传递DEN-1、4到子3代。传递DEN-3到子2代，白纹伊蚊兴义株能垂直传递DEN-2到子3代，白纹伊蚊贵阳．株能垂直传递DEN-1到子3代，传递DEN-2、3、4到子2代，白纹伊蚊毕节株能垂直传递DEN-1到子2代，DEN-4至子3代，未发现能垂直传递DEN-2、3。结论对DEN1—4易感的白纹伊蚊贵阳、兴义、毕节株能垂直传递DEN，且对DEN的垂直传递能力与其地理来源有关；贵州省3个地方株的白纹伊蚊卵可在干燥室温条件下越冬保存DEN。

Study on vertical transmission with dengue virus in Aedes albopietus

Objective To study vertical transmission capability of dengue virus type 1-4(DEN 1-4) through three geographic strains Aedes albopictus in Guizhou province.Methods Three geographic strains of Aedes albopictus in Guizhou Province were infected orally with DEN1-4,respectively.Parental and F1-F3 generation mosquitoes were gathered.Dengue virus was isolated with C6/36;The viral antigen was detected by indirect immunofluore sences assay(IFA).The total RNA was obtained,NS1 gene of DEN was detected by RT-PCR.Results Three Guizhou geographic mosquitoes of Aedes albopictus could be infected orally with DEN1-4;Both Xingyi and Hainan geographic mosquitoes had the capbility of vertical transmission DEN-1 and DEN-4 to F3 generation and DEN-3 to F2 generation as well,Xingyi geographic mosquitoes were detected vertical transmission DEN-2 to F3 generation;Guiyang geographic mosquitoes could vertically transmit DEN-1 to F3 generation,DEN-2,DEN-3 and DEN-4 to F2 generation;Transmitted DEN-1 to F2 generation,DEN-4 to F3 generation,transmitted DEN-2 and DEN-3 to progeny were not be detected by Bijie mosquitoes.Conclusion Three geographic strains Aedes albopictus in Guizhou were susceptible to dengue virus type 1-4;Guiyang,Bijie and Xingyi geographic mosquitoes of Aedes Aalbopictus could vertical transmit DEN,and their efficiency were various.The eggs of filial mosquitoes could live through the winter at dry,room temperature.