Molecular and microscopic analysis of bacteria and viruses in exhaled breath collected using a simple impaction and condensing method.

Exhaled breath condensate (EBC) is increasingly being used as a non-invasive method for disease diagnosis and environmental exposure assessment. By using hydrophobic surface, ice, and droplet scavenging, a simple impaction and condensing based collection method is reported here. Human subjects were...

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Autores principales:	Zhenqiang Xu, Fangxia Shen, Xiaoguang Li, Yan Wu, Qi Chen, Xu Jie, Maosheng Yao
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/f3c5f36ea800418bba13769238e8946f
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spelling	oai:doaj.org-article:f3c5f36ea800418bba13769238e8946f2021-11-18T07:11:10ZMolecular and microscopic analysis of bacteria and viruses in exhaled breath collected using a simple impaction and condensing method.1932-620310.1371/journal.pone.0041137https://doaj.org/article/f3c5f36ea800418bba13769238e8946f2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22848436/?tool=EBIhttps://doaj.org/toc/1932-6203Exhaled breath condensate (EBC) is increasingly being used as a non-invasive method for disease diagnosis and environmental exposure assessment. By using hydrophobic surface, ice, and droplet scavenging, a simple impaction and condensing based collection method is reported here. Human subjects were recruited to exhale toward the device for 1, 2, 3, and 4 min. The exhaled breath quickly formed into tiny droplets on the hydrophobic surface, which were subsequently scavenged into a 10 µL rolling deionized water droplet. The collected EBC was further analyzed using culturing, DNA stain, Scanning Electron Microscope (SEM), polymerase chain reaction (PCR) and colorimetry (VITEK 2) for bacteria and viruses.Experimental data revealed that bacteria and viruses in EBC can be rapidly collected using the method developed here, with an observed efficiency of 100 µL EBC within 1 min. Culturing, DNA stain, SEM, and qPCR methods all detected high bacterial concentrations up to 7000 CFU/m(3) in exhaled breath, including both viable and dead cells of various types. Sphingomonas paucimobilis and Kocuria variants were found dominant in EBC samples using VITEK 2 system. SEM images revealed that most bacteria in exhaled breath are detected in the size range of 0.5-1.0 µm, which is able to enable them to remain airborne for a longer time, thus presenting a risk for airborne transmission of potential diseases. Using qPCR, influenza A H3N2 viruses were also detected in one EBC sample. Different from other devices restricted solely to condensation, the developed method can be easily achieved both by impaction and condensation in a laboratory and could impact current practice of EBC collection. Nonetheless, the reported work is a proof-of-concept demonstration, and its performance in non-invasive disease diagnosis such as bacterimia and virus infections needs to be further validated including effects of its influencing matrix.Zhenqiang XuFangxia ShenXiaoguang LiYan WuQi ChenXu JieMaosheng YaoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 7, p e41137 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Zhenqiang Xu Fangxia Shen Xiaoguang Li Yan Wu Qi Chen Xu Jie Maosheng Yao Molecular and microscopic analysis of bacteria and viruses in exhaled breath collected using a simple impaction and condensing method.
description	Exhaled breath condensate (EBC) is increasingly being used as a non-invasive method for disease diagnosis and environmental exposure assessment. By using hydrophobic surface, ice, and droplet scavenging, a simple impaction and condensing based collection method is reported here. Human subjects were recruited to exhale toward the device for 1, 2, 3, and 4 min. The exhaled breath quickly formed into tiny droplets on the hydrophobic surface, which were subsequently scavenged into a 10 µL rolling deionized water droplet. The collected EBC was further analyzed using culturing, DNA stain, Scanning Electron Microscope (SEM), polymerase chain reaction (PCR) and colorimetry (VITEK 2) for bacteria and viruses.Experimental data revealed that bacteria and viruses in EBC can be rapidly collected using the method developed here, with an observed efficiency of 100 µL EBC within 1 min. Culturing, DNA stain, SEM, and qPCR methods all detected high bacterial concentrations up to 7000 CFU/m(3) in exhaled breath, including both viable and dead cells of various types. Sphingomonas paucimobilis and Kocuria variants were found dominant in EBC samples using VITEK 2 system. SEM images revealed that most bacteria in exhaled breath are detected in the size range of 0.5-1.0 µm, which is able to enable them to remain airborne for a longer time, thus presenting a risk for airborne transmission of potential diseases. Using qPCR, influenza A H3N2 viruses were also detected in one EBC sample. Different from other devices restricted solely to condensation, the developed method can be easily achieved both by impaction and condensation in a laboratory and could impact current practice of EBC collection. Nonetheless, the reported work is a proof-of-concept demonstration, and its performance in non-invasive disease diagnosis such as bacterimia and virus infections needs to be further validated including effects of its influencing matrix.
format	article
author	Zhenqiang Xu Fangxia Shen Xiaoguang Li Yan Wu Qi Chen Xu Jie Maosheng Yao
author_facet	Zhenqiang Xu Fangxia Shen Xiaoguang Li Yan Wu Qi Chen Xu Jie Maosheng Yao
author_sort	Zhenqiang Xu
title	Molecular and microscopic analysis of bacteria and viruses in exhaled breath collected using a simple impaction and condensing method.
title_short	Molecular and microscopic analysis of bacteria and viruses in exhaled breath collected using a simple impaction and condensing method.
title_full	Molecular and microscopic analysis of bacteria and viruses in exhaled breath collected using a simple impaction and condensing method.
title_fullStr	Molecular and microscopic analysis of bacteria and viruses in exhaled breath collected using a simple impaction and condensing method.
title_full_unstemmed	Molecular and microscopic analysis of bacteria and viruses in exhaled breath collected using a simple impaction and condensing method.
title_sort	molecular and microscopic analysis of bacteria and viruses in exhaled breath collected using a simple impaction and condensing method.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/f3c5f36ea800418bba13769238e8946f
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Molecular and microscopic analysis of bacteria and viruses in exhaled breath collected using a simple impaction and condensing method.

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