嗜肺军团菌荧光定量PCR检测方法的建立_张琦

了采样的效率。三是混杂物质和环境的干扰,国外的

研究发现空气中细菌和真菌等混杂物质的干扰会降低撞击法的采样效率,通过添加高效过滤网,可以提高采样效率[12]。此外环境的温湿度对流感病毒的采集效率也有一定的影响[13]

。

本实验证实了使用新型静电场采样装置采集分析流感病毒是可行性的,可以进一步改进目前的采样器,通过延长气溶胶在采样器中的停留时间,提高其采集的效率。

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・专题论著・

嗜肺军团菌荧光定量PCR 检测方法的建立

张琦1,陈晓东1,2,张宝莹3,钱乐4,司朝宗4,张秀珍2,甄世祺2,刘凡3,陈连生2

(1.南京医科大学公共卫生学院,　江苏南京　210029;2.江苏省疾病预防控制中心,　江苏南京　210009;

3.中国疾病预防控制中心环境与健康相关产品安全所,　北京　100050;

4.东南大学公共卫生学院,　江苏南京　210009)

DOI :10.3969/j.issn.1006-9070.2010.03.002

基金项目:“十一五”国家科技支撑计划项目(2006BAI19B04)收稿日期:2010-03-22

作者简介:张琦(1984—

),女,内蒙古赤峰人,硕士研究生在读。 【摘　要】　目的:建立嗜肺军团菌mip 基因荧光定量PCR 检测方法。方法:利用Primer Express 软件设计特异性引物和探针,对质粒标准品、嗜肺军团菌、非嗜肺军团菌及其他菌株进行检测,验证方法的特异性、敏感性和重复性,最后采集环境样本进行检测。结果:该方法特异性好,嗜肺军团菌呈现阳性结果,而非嗜肺军团菌及其他菌株均为阴性结果。检测灵敏度达

293copies/μl ;重复性较好,Ct 值变异系数较小;检测的12份环境样本中5份阳性。结论:该方法快速且具有较好的特异性、

敏感性、重复性,适于外环境嗜肺军团菌污染调查及应急事件的快速检测。

【关键词】　嗜肺军团菌;荧光定量PCR 检测;mip 基因

【中图分类号】　R -331　【文献标识码】　A 【文章编号】　1006-9070(2010)03-0003-04