• 关键词：复制缺陷型重组腺病毒  气溶胶  实时荧光定量PCR  绿色荧光

• 基金项目：国家自然科学基金重点项目(No40332024)

• 李晓岩
• 广州医学院实验医学研究中心, 广州 510182

• 刘启才
• 广州医学院实验医学研究中心, 广州 510182

• 彭燕
• 广州医学院实验医学研究中心, 广州 510182

• 毕新慧
• 中国科学院广州地球化学研究所有机地球化学国家重点实验室, 广州 510640

• 李冰
• 广州医学院实验医学研究中心, 广州 510182

• 摘要：将带有绿色荧光蛋白的复制缺陷型重组腺病毒,作为模拟病毒建立一种检测病毒侵袭力的方法.在钢化玻璃箱中通过TK-3型微生物气溶胶发生器将腺病毒形成气溶胶,用FA-1型多级撞击式空气微生物采样器进行气溶胶采样,对采样样品分别进行实时荧光定量PCR检测和表达了绿色荧光蛋白的PK15活细胞定时检测.实时荧光定量PCR检测可测定病毒在大气中存在的相对基因拷贝数,通过在荧光显微镜下计数带绿色荧光的PK15细胞数可直观检测病毒的感染力及活力.结果表明,重组腺病毒气溶胶主要分布在采样器第五级,腺病毒气溶胶与病毒粒子相比较大.

• Abstract：Replication deficient recombinant adenovirus carrying green fluorescent protein (GFP) was used as a model virus to establish a method to analyze the invasiveness of virus aerosols in vital cells. An adenovirus aerosol was generated using a TK-3 aerosol generator attached to a stalinite chamber. Virus aerosol specimens were collected with a FA-1 six-stage impact sampler. The relative genome copy number of viruses in the aerosol were detected by real-time fluorescence quantitative PCR. The number of virus-infected PK15 cells was determined by counting cells with green fluorescence under a fluorescence microscope. The PCRresults showed that viral DNAexisted in stages 3～6 of the sampler, corresponding to aerosols 1.1～ 3.3μm in size, and was most abundant in stage 5. Cells with green fluorescence were also found in stages 3～6, and were most abundant in stage 5. These results provide evidence that the viruses are present in the atmosphere as virus aerosols, which are much larger than their own particle size. Whether the virus distribution in the aerosol is affected by atmospheric conditions,and the relationship between transmission of virus and atmospheric conditions will be an important issue of investigation.

• Key words：Replication deficient recombinant adenovirus  aerosol  real-time fluorescence quantitative PCR  green fluorescence

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