云南登革4型病毒的鉴定及NS1和NS2a基因序列分析  

Identification and sequence analysis of NS1 and NS2a of two strains of dengue type-4 virus from mosquitoes in Yunnan province

作　　者：王静林[1] 张海林[1] 孙肖红[2] 付士红[2] 米竹青[1] 龚正达[1] 翟友刚[2] 唐青[2] 梁国栋[2] 

机构地区：[1]云南省地方病防治所／云南省病毒立克次体研究中心,大理671000 [2]中国疾病预防控制中心病毒病预防控制所,北京100052

出　　处：《中国人兽共患病学报》2008年第7期636-640,共5页Chinese Journal of Zoonoses

基　　金：国家自然科学基金资助项目(No.30560142)

摘　　要：目的对1989年8月分离自云南省盈江县蚊虫的2株登革病毒进行生物学及分子特征的鉴定,明确这两株病毒的血清型及基因型。方法蚊虫用BHK21细胞和乳鼠法分离病毒,用间接免疫荧光试验、RT-PCR等方法进行鉴定,并对这两株病毒的分子生物学特征进行分析。结果从白纹伊蚊和达勒姆阿蚊中各分离到一株病毒,分别命名为M110和M113,这两株病毒对BHK21细胞能产生明显的细胞病变,脑内接种乳鼠4d左右导致乳鼠死亡。该病毒经血凝、血凝抑制和间接免疫荧光试验提示为黄病毒。分别用黄病毒属特异引物、登革4型病毒NS1和NS2a基因片段特异引物进行RT-PCR扩增、序列测定,证实为登革4型病毒。NS1和NS2a基因序列片段分析表明,M110和M113的NS1核苷酸序列与登革4型病毒基因Ⅰ型的H241株(Y19176)同源性最高,分别为99%、98%;NS2a基因片段与H241的核苷酸序列的同源性分别为96.5%、97.1%。结论从盈江县蚊虫分离到的M110和M113病毒为登革4型病毒基因Ⅰ型,表明云南省西部边境地区在20世纪80年代发生过登革4型病毒的流行。To identify the serum type and genotype of two dengue virus strains M110 and M113 from mosquitoes collected in 1989 in Yinjiang county, Yunnan province of China. BHK21 cell line and 3-day suckling mice were inoculated with two viruses and molecular biological characteristics of two viruses were identified by indirect immunofluorescent antibody and reverse transcription-polymerase chain reaction （RT-PCR） techniques. Strains M110 and M113 isolated from Aedes albopictus and Armigeres durhami, respectively, which were fatal to suckling mice at 4 days alter intracerebrally inoculation and had eytopathologic effects on BHK21 cells, showed a positive reaction to dengue antibody and could be amplified Ly using flavivirus primers and dengue type-4 NSl gene primers and dengue type-4 NS2a gene primers. The nucleotide sequences showed that compared with dengue type-4 virus gene type I （H241 strain,Y19176）, NSl sequences of M110 and M113 had a very high homology to H241 （99% and 98%） and the similar results with NS2a（96.5% and 97.1%）. The isolated 2 virus strains belonged to gene type I of dengue type-4 virus and such results showed that there existed epidemic probabilities of the disease in western border region el Yunnan province in last 1980＇ s.

关 键 词：登革4型病毒 NS1 NS2a 基因型 

分 类 号：R512.82[医药卫生—内科学]