使用管家基因进行生物气溶胶组分分析的初步研究
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摘要
空气中生物成分来源多样,对人类健康构成潜在威胁,目前缺少有效监测空气中不同生物成分含量的技术,通过检测不同物种的管家基因进行鉴别是可选的手段。本研究使用BIO Capturer-6病毒气溶胶采集富集仪收集环境样本28份,分别使用细菌、动物和植物管家基因特异性引物探针进行荧光定量PCR扩增,检测分析空气中各物种含量差异。结果表明:全部样本均检出细菌、动物和植物成分;不同高度(地面和楼顶)空气中各生物组分含量无统计学差异,温度高、湿度小时,空气中细菌成分含量最多。本研究创造性地将管家基因与生物气溶胶检测结合起来,建立了评价空气质量的新方法;并通过实验证明了该方法有良好的稳定性和灵敏度,相信可以为空气生物学调查分析提供新的手段。
The complexity of biological components in the air poses a potential threat to human health.There is currently no effective technology to monitor the content of different biological components in the air. Examining the housekeeping genes of different species is an optional means to identify these compo-nents. Herein, 28 environmental samples were collected using the BIO Capturer-6, and the real-time PCR amplification was performed using bacterial, animal and plant housekeeping gene primer probes to detect and analyze the differences in the content of each species. The results showed that bacterial, animal and plant components were detectable in all the samples, and that the contents of various biological components in the air at different heights(ground and roof) were not statistically different. The bacterial component was the most in the case of both high temperature and low humidity. This research creatively combined housekeeping genes with bioaerosol detection for evaluating air quality. It proved that the method has a good stability and sensitivity and could be used for the investigation and analysis of aerobiology.
The complexity of biological components in the air poses a potential threat to human health.There is currently no effective technology to monitor the content of different biological components in the air. Examining the housekeeping genes of different species is an optional means to identify these compo-nents. Herein, 28 environmental samples were collected using the BIO Capturer-6, and the real-time PCR amplification was performed using bacterial, animal and plant housekeeping gene primer probes to detect and analyze the differences in the content of each species. The results showed that bacterial, animal and plant components were detectable in all the samples, and that the contents of various biological components in the air at different heights(ground and roof) were not statistically different. The bacterial component was the most in the case of both high temperature and low humidity. This research creatively combined housekeeping genes with bioaerosol detection for evaluating air quality. It proved that the method has a good stability and sensitivity and could be used for the investigation and analysis of aerobiology.
引文
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[16]刘杨,马雪征,张丽萍,等.生物气溶胶采集富集仪用于医院空气中甲型流感病毒的调查[J].中国国境卫生检疫杂志(LIUYang,MA Xue-zheng,ZHANG Li-ping,et al.Survey on influenza a virus in hospital air by using biological aerosol collection and enrichment instrument[J].Chinese Journal of Frontier Health and Quarantine),2018,41(1):5-7.
[17]祁建华,武丽婧,高冬梅,等.青岛近海生物气溶胶中可培养微生物浓度及群落多样性的季节变化[J].环境科学(QI Jianhua,WU Li-jing,GAO Dong-mei,et al.Concentration and community diversity of microbes in bioaerosols in the Qingdao coastal region[J].Environmental Science),2014,35(3):801-809.
[18]李艾阳,东东,孟志航,等.典型高校公共场所室内空气微生物分布特征[J].环境科学学报(LI Ai-yang,DONG Dong,MENGZhi-hang,et al.Distribution of indoor air microorganisms in public rooms of a university[J].Acta Scientiae Circumstantiae),2016,36(8):2827-2831.
[19]杨永存,杨冬燕,李浩,等.实时荧光PCR检测动物源性基因鉴定地沟油[J].中国卫生检验杂志(YANG Yong-cun,YANGDong-yan,LI Hao,et al.Identification of hogwash oil through animal-original genes detection with real-time PCR[J].Chinese Journal of Health Laboratory Technology),2013,23(18):3514-3517.
[20]陈颖,吴亚君,袁飞,等.GB/T 25165-2010,明胶中牛、羊、猪源性成分的定性检测方法---实时荧光PCR法[S].中国检验检疫科学研究院(CHEN Ying,WU Ya-jun,YUAN Fei,et al.GB/T 25165-2010,Protocol of identification of bovine,caprine,ovine and porcine derived materials in gelation-Real time PCRmethod[S].Chinese Academy of Inspection and Quarantine).2010-09-26.
[21]岳巧云,陈定虎,伍朝晖,等.实时荧光PCR在鉴别奶粉中掺入大豆成分的应用研究[J].食品科学(YUE Qiao-yun,CHENDing-hu,WU Zhao-hui,et al.Real-time fluorescent PCR detection of soybean ingredients adulterate in milk powder[J].Food Science),2009,30(12):190-193.
[2]杜睿.大气生物气溶胶的研究进展[J].气候与环境研究(DU Rui.The progress of atmospheric bioaerosol research[J].Climatic and Environmental Research),2006,11(4):546-552.
[3]HAAS D,GALLER H,LUXNER J,et al.The concentrations of culturable microorganisms in relation to particulate matter in urban air[J].Atmospheric Environment,2013,65(2):215-222.
[4]江桂斌.快速发展的生物气溶胶学科[J].科学通报(JIANG Guibin.Rapid development of bioaerosol science[J].Chinese Science Bulletin),2018,63(10):875.
[5]ZHANG L,ZHANG Q,JIANG Y,et al.Reference genes identification for normalization of qPCR under multiple stresses in Hordeum brevisubulatum[J].Plant Methods,2018,14:110.
[6]万松华.食品中污染微生物通用型荧光PCR检测方法研究[D].广州:华南理工大学(WAN Song-hua.A Study of Detecting Microorganism in Food by Real-time PCR Method with Universal Primers and Probe[D].Guangzhou:South China University of Technology),2015.
[7]赵新,易萌,刘娜,等.DB12/T 653-2016,鲜冻畜、禽肉中动物源性成分的定量检测---实时荧光PCR法[S].天津市农业质量标准与检测技术研究所,天津市东丽区产品质量监督检验所(ZHAO Xin,YI Meng,LIU Na,et al.DB12/T 653-2016,Quantitative PCR method for animal derived materials in meat and meat products[S].Tianjin Institute of Agricultural Quality Standards and Testing Technology,Tianjin Dongli District Product Quality Supervision and Inspection Institute).2016-09-27.
[8]顾晓慧,姚琳,王联珠,等.冷冻鱼糜中植物成分的PCR检测方法[J].中国渔业质量与标准(GU Xiao-hui,YAO Lin,WANGLian-zhu,et al.PCR detection method of plant ingredient in frozen surimi[J].Chinese Fishery Quality and Standards),2014,4(2):44-49.
[9]KESAVAN J,SAGRIPANTI J L.Evaluation criteria for bioaerosol samplers[J].Environmental Science.Processes&Impacts,2015,17(3):638-645.
[10]MAMTA,SHRIVASTAVA J N,SATSANGI G P,et al.Assessment of bioaerosol pollution over Indo-Gangetic plain[J].Environmental Science and Pollution Research International,2015,22(8):6004-6009.
[11]刘民,马秋月.肺炎链球菌相关疾病流行病学研究进展[J].中国公共卫生(LIU Min,MA Qiu-yue.Progress in epidemiological studies on pneumococcal diseases[J].China Journal of Public Health),2018,34(11):1449-1452.
[12]宋柏龙,孙勇,许栋岳,等.蒿属花粉分布及对过敏性鼻炎患者气道反应性的影响分析[J].中国医学文摘(耳鼻咽喉科学)(SONG Bo-long,SUN Yong,XU Dong-yue,et al.Distribution of artemisia pollen and its effect on airway responsiveness in patients with allergic rhinitis[J].Chinese Medical Digest:Otorhinolaryngology),2018,33(5):385-387.
[13]孙凌聪,吴冬妮,张华勋,等.湖北省首例输入性疟疾混合感染的实验室诊断分析[J].公共卫生与预防医学(SUN Lingcong,WU Dong-ni,ZHANG Hua-xun,et al.Laboratory diag nosis of the first mixed infection of imported malaria in Hubei[J].Journal of Public Health and Preventive Medicine),2017,28(2):131-133.
[14]刘起勇.寨卡病毒媒介伊蚊控制策略和措施展望[J].中国媒介生物学及控制杂志(LIU Qi-yong.Prospects on Zika virus control measures and strategy[J].Chinese Journal of Vector Biology and Control),2016,27(2):93-98.
[15]高敏,仇天雷,贾瑞志,等.北京雾霾天气生物气溶胶浓度和粒径特征[J].环境科学(GAO Min,QIU Tian-lei,JIA Rui-zhi,et al.Concentration and size distribution of bioaerosols at nonhaze and haze days in Beijing[J].Environmental Science),2014,35(12):4415-4421
[16]刘杨,马雪征,张丽萍,等.生物气溶胶采集富集仪用于医院空气中甲型流感病毒的调查[J].中国国境卫生检疫杂志(LIUYang,MA Xue-zheng,ZHANG Li-ping,et al.Survey on influenza a virus in hospital air by using biological aerosol collection and enrichment instrument[J].Chinese Journal of Frontier Health and Quarantine),2018,41(1):5-7.
[17]祁建华,武丽婧,高冬梅,等.青岛近海生物气溶胶中可培养微生物浓度及群落多样性的季节变化[J].环境科学(QI Jianhua,WU Li-jing,GAO Dong-mei,et al.Concentration and community diversity of microbes in bioaerosols in the Qingdao coastal region[J].Environmental Science),2014,35(3):801-809.
[18]李艾阳,东东,孟志航,等.典型高校公共场所室内空气微生物分布特征[J].环境科学学报(LI Ai-yang,DONG Dong,MENGZhi-hang,et al.Distribution of indoor air microorganisms in public rooms of a university[J].Acta Scientiae Circumstantiae),2016,36(8):2827-2831.
[19]杨永存,杨冬燕,李浩,等.实时荧光PCR检测动物源性基因鉴定地沟油[J].中国卫生检验杂志(YANG Yong-cun,YANGDong-yan,LI Hao,et al.Identification of hogwash oil through animal-original genes detection with real-time PCR[J].Chinese Journal of Health Laboratory Technology),2013,23(18):3514-3517.
[20]陈颖,吴亚君,袁飞,等.GB/T 25165-2010,明胶中牛、羊、猪源性成分的定性检测方法---实时荧光PCR法[S].中国检验检疫科学研究院(CHEN Ying,WU Ya-jun,YUAN Fei,et al.GB/T 25165-2010,Protocol of identification of bovine,caprine,ovine and porcine derived materials in gelation-Real time PCRmethod[S].Chinese Academy of Inspection and Quarantine).2010-09-26.
[21]岳巧云,陈定虎,伍朝晖,等.实时荧光PCR在鉴别奶粉中掺入大豆成分的应用研究[J].食品科学(YUE Qiao-yun,CHENDing-hu,WU Zhao-hui,et al.Real-time fluorescent PCR detection of soybean ingredients adulterate in milk powder[J].Food Science),2009,30(12):190-193.