河北省汉坦病毒的分离、复苏及分型

发布时间：2018-06-12 06:48

  本文选题：汉坦病毒 + 细胞培养　； 参考：《中国媒介生物学及控制杂志》2016年05期

【摘要】：目的对2012年河北省鼠类标本进行汉坦病毒(HV)分离并复苏保存的病毒株,鉴定型别,了解河北省疫区HV的型别分布特点。方法利用间接免疫荧光法(IFA)对鼠肺标本进行初筛,采用细胞培养法分离阳性标本病毒,并复苏已保存的病毒株。提取病毒RNA后进行分型鉴定。结果 4份阳性鼠肺标本的组织悬液接种Vero-E6细胞2~3代后,经IFA检测,细胞内出现病毒特异性荧光颗粒,但未发现致细胞病变效应。4份标本全部分离成功。取已保存的病毒株接种Vero-E6细胞1~2代后,经IFA检测,细胞内出现较强的病毒特异性荧光颗粒,6株病毒全部复苏成功。通过分型鉴定,10株病毒均属于家鼠型汉城型汉坦病毒(SEOV)。结论河北省HV以SEO型为主。SEOV在Vero-E6细胞虽无明显的细胞病变,但可在该细胞中增殖并被检测。在-80℃条件下,HV至少保存20余年。我国东方田鼠为SEOV的宿主之一。
[Abstract]:Objective to isolate and resuscitate the strain of Hantavirus (HVV) from rat samples in Hebei Province in 2012, to identify the type of HV and to understand the distribution characteristics of HV in epidemic area of Hebei Province. Methods indirect immunofluorescence assay (IFA) was used to screen the lung samples of mice. The positive specimens were isolated by cell culture and the preserved virus strains were resuscitated. The virus RNA was extracted and identified. Results Vero-E6 cells were inoculated with the tissue suspensions of 4 positive mice lung specimens for 2 ~ 3 passages. After IFA detection, virus specific fluorescent particles were found in the cells, but none of the 4 specimens were isolated successfully. After the preserved virus strain was inoculated into Vero-E6 cells for 1 and 2 passages, the results of IFA showed that all of the 6 virus strains with strong virus specific fluorescence particles were successfully resuscitated. It was identified by typing that all 10 strains belonged to the Seoul type of Hantavirus of domestic mouse type SEOVV. Conclusion HV of Hebei Province is mainly SEO type. SEOV has no obvious cytopathic effect in Vero-E6 cells, but it can proliferate and be detected in Vero-E6 cells. HV was preserved at -80 鈩，

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