空气中流感病毒富集于采样介质中的存活情况与监测应用

Detection of the Survival of the Influenza Viruses Enriched in the Sampling Media and Applications of the Sampling Methods in Air Monitoring

目的建立流感病毒气溶胶富集方法,检测富集病毒气溶胶存活情况并评价空气微生物采样器的采样效率。方法通过气溶胶发生器模拟空气中流感病毒,使用30μm孔径超细静电纤维膜、0.2μm孔径Teflon静电膜和AGI-30大气采样器分别对流感病毒进行富集监测,运用荧光定量PCR和MDCK细胞培养方法,分析探讨不同采样介质中流感病毒存活情况与采样效率。结果在一定的条件下富集空气中的流感病毒,超细静电纤维膜和Teflon膜收集到的病毒存活率低,而使用7%胎牛血清处理过的超细静电纤维膜和Teflon膜采样,则病毒存活率高;使用静电膜富集空气中的流感病毒,疫点和医院发热门诊流感病毒RNA阳性检出率分别为100%和20%,而使用AGI-30采样病毒RNA检测全为阴性。结论监测空气中流感病毒的活性是有效预防流感的关键,空气中流感病毒气溶胶的富集,采用较大孔径和小孔径的静电膜都具有较高的富集效能,可应用于空气环境监测。

Objective To establish a method to enrich the influenza viruses in aerosol, and to detect the survival of the enriched viruses and to evaluate the sampling efficiency of the microbial air sampler. Methods Influenza viruses were simulat- ed into the air through the aerosol generator. The viruses were concentrated by ultrafine electrostatic fiber membrane （aperture size = 30μm）, teflon electrostatic membrane （aperture size= 0.2μm） and the AGI - 30 atmosphere sampler separately. Then the viruses were detected by fluorescence PCR and MDCK - cell culture. The survival of the viruses collected by different sampling media were detected and the sampling efficiency was analyzed. Results The survival rates of the influenza viruses concentrat- ed by ultrafine electrostatic fiber membrane and teflon electrostatic membrane were low. However, after these two membranes were pretreated by 7 % fetal bovine serum, the survival rates were increased. When the air influenza viruses in the epidemic focus and the fever clinic of a hospital were enriched by the electrostatic fiber membrane, the detection rates of the influenza viral RNA were 100 % and 20 % respectively, while when the viruses were concentrated by the AGI - 30 atmosphere sampler, the influenza viral RNA was not detected. Conclusions For acquisition of influenza viruses in the air, the electrostatic membrane of both the large and small aperture size have higher enrichment efficiency and can be used in air monitoring. However, the enrichment efficiency of the AGI - 30 atmosphere sampler is low and should not be used for a long time with a high flow rate sampling.