吉林省普马拉病毒的RT-PCR检测与序列分析

RT-PCR Detection and Sequence Analysis of Puumala Virus in Jilin Province

目的建立检测普马拉病毒的RT—PCR方法，确认吉林省存在普马拉病毒，并对病毒的基因片段进行分析。方法采用RT—PCR方法，对吉林省抚松地区棕背（[鼠平]）鼠肺标本进行检测。对S片段核蛋白基因编码序列进行序列比较分析，绘制系统发生树。结果研究表明在吉林省抚松地区存在普马拉病毒，采集于2005年和2006年的标本阳性率分别为7．34％和14．29％，呈逐年上升趋势。序列分析表明与其他普马拉病毒核苷酸序列同源性只有81．1％～83．3％。系统发生分析揭示，该病毒与在日本、韩国分离的普马拉病毒亲缘关系相对接近，并且在进化过程中处于更古老的节点。结论在吉林省抚松地区可能存在普马拉病毒的一个新亚型。

Objective To establish RT - PCR method for Puumala virus（PUUV） detection in order to confirm existence of Puumala virus in Jilin province and analyze gene sequence of Puumala virus. Methods A method based on RT - PCR was used for the detection of Puumala virus in lung samples of Clethrionomys rufocanas trapped in Fusong district of Jilin province. Sequence analysis of nucleocapsid protein coding sequences among S segment of Puumala virus was performed and phylogenetic tree was constructed. Results The research showed the existence of Puumala virus in Fusong district, the positive rate was 7. 34% for the samples collected in 2005, and 14. 29% in 2006, which showed a increment trend year after year. Sequence analysis showed that the nucleotide homology between Fusong virus and other PUUV was 81.1% - 83.3%. Phylogenetic analysis revealed that the Puumala virus found in Fusong was related to the isolates from Japan and Korea, and occupied more ancestral node in the evolution. Conclusion A new subtype of Puumala virus is probably existed in Fusong district of Jilin province.