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腺病毒气溶胶的实时荧光定量PCR检测和绿色荧光蛋白活细胞检测
  • ISSN号:0253-2468
  • 期刊名称:《环境科学学报》
  • 时间:0
  • 分类:X830.2[环境科学与工程—环境工程]
  • 作者机构:[1]广州医学院实验医学研究中心,广州510182, [2]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
  • 相关基金:国家自然科学基金重点项目(No.40332024)
中文摘要:

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.

英文摘要:

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-tlme fluorescence quantitative PCR. The number of virus-infected PK15 cells was determined by counting cells with green fluorescence under a fluorescence microscope. The PCR results showed that viral DNA existed in stages 3 - 6 of the sampler, corresponding to aerosols 1.1 - 3.31μm 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.

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期刊信息
  • 《环境科学学报》
  • 中国科技核心期刊
  • 主管单位:中国科学院
  • 主办单位:中国科学院生态环境研究中心
  • 主编:汤鸿霄
  • 地址:北京2871信箱
  • 邮编:100085
  • 邮箱:hjkxxb@rcees.ac.cn
  • 电话:010-62941073
  • 国际标准刊号:ISSN:0253-2468
  • 国内统一刊号:ISSN:11-1843/X
  • 邮发代号:82-625
  • 获奖情况:
  • 国内外数据库收录:
  • 美国化学文摘(网络版),荷兰地学数据库,荷兰文摘与引文数据库,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),英国英国皇家化学学会文摘,中国北大核心期刊(2000版)
  • 被引量:56074