应用单引物差异RT-PCR法鉴定我国分离的甲病毒(YN87448病毒)

甲病毒指披膜病毒科(Togavirdae)甲病毒属(Alphavirus),以蚊虫等吸血昆虫为传播媒介,可引起如辛德毕斯热、基孔肯雅热、东方马脑炎、西方马脑炎等多种人畜共患性传染病1,2],是医学上重要的虫媒病毒.甲病毒世界性分布,全世界已发现28种甲病毒,其中13种与人畜疾病有关,至少7种甲病毒发生过不同程度流行,造成巨大的经济损失,成为严重的公共卫生问题.张海林等(中国媒介生物学及控制杂志,1992,3(特7):419－421)从云南傣族一位发热病人血中分离到一株病毒,命名为YN87448病毒.经血清学鉴定该病毒符合披膜病毒科甲病毒属病毒特征.由于缺乏标准诊断血清,无法对它作进一步鉴定.该病毒直接分离自发热病人血清,病人主要症状为发热,寒颤,腰疼痛和全身酸痛.由于该病毒直接分离自病人血清,病毒鉴定对解释该病人的发病原因,甚至对于解释我国部分地区流行的"无名热"的病因均具有重要意义.

Identification of Alphavirus YN87448 Strain Using Single-primer Differential RT-PCR

YN87448 virus strain was isolated from a fevered female patient (52 years old ) in Yunnan Province in 1986, and was identified as a member of Alphavirus using serological method. One primer was designed from common hairpin conserved region of Alphavirus. Two fragments were amplified by single-primer differential RT-PCR, and they were cloned into pGEM-T vector. Sequences were determined, and then analyzed by Software and GenBank. Results showed that the 1118pb long fragment was highly homologous to the sequence of Sindbis-like virus S.A.A.R86. The homogeneity of the 1118bp fragment between YN87448 virus strain and S.A.A.R86 virus strain was 98%. Single-primer differential RT-PCR is a useful method with simple, economic, and practical value for Alphavirus identification.