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	Quantification of viable bacteria in wastewater treatment plants by using propidium monoazide combined with quantitative PCR(PMA-qPCR)
	Dan Li ; Tiezheng Tong ; Siyu Zeng ; Yiwen Lin ; Shuxu Wu ; Miao He ; Dan Li ; Tiezheng Tong ; Siyu Zeng ; Yiwen Lin ; Shuxu Wu ; Miao He
	2016-03-30 ; 2016-03-30
关键词	propidium monoazide antitative PCR TPs E.coli terococci X832 X703
其他题名	Quantification of viable bacteria in wastewater treatment plants by using propidium monoazide combined with quantitative PCR(PMA-qPCR)
中文摘要	The detection of viable bacteria in wastewater treatment plants(WWTPs) is very important for public health, as WWTPs are a medium with a high potential for waterborne disease transmission. The aim of this study was to use propidium monoazide(PMA) combined with the quantitative polymerase chain reaction(PMA-qPCR) to selectively detect and quantify viable bacteria cells in full-scale WWTPs in China. PMA was added to the concentrated WWTP samples at a final concentration of 100 μmol/L and the samples were incubated in the dark for 5 min, and then lighted for 4 min prior to DNA extraction and qPCR with specific primers for Escherichia coli and Enterococci, respectively. The results showed that PMA treatment removed more than 99% of DNA from non-viable cells in all the WWTP samples, while matrices in sludge samples markedly reduced the effectiveness of PMA treatment. Compared to qPCR, PMA-qPCR results were similar and highly linearly correlated to those obtained by culture assay, indicating that DNA from non-viable cells present in WWTP samples can be eliminated by PMA treatment, and that PMA-qPCR is a reliable method for detection of viable bacteria in environmental samples. This study demonstrated that PMA-qPCR is a rapid and selective detection method for viable bacteria in WWTP samples, and that WWTPs have an obvious function in removing both viable and non-viable bacteria. The results proved that PMA-qPCR is a promising detection method that has a high potential for application as a complementary method to the standard culture-based method in the future.; The detection of viable bacteria in wastewater treatment plants(WWTPs) is very important for public health, as WWTPs are a medium with a high potential for waterborne disease transmission. The aim of this study was to use propidium monoazide(PMA) combined with the quantitative polymerase chain reaction(PMA-qPCR) to selectively detect and quantify viable bacteria cells in full-scale WWTPs in China. PMA was added to the concentrated WWTP samples at a final concentration of 100 μmol/L and the samples were incubated in the dark for 5 min, and then lighted for 4 min prior to DNA extraction and qPCR with specific primers for Escherichia coli and Enterococci, respectively. The results showed that PMA treatment removed more than 99% of DNA from non-viable cells in all the WWTP samples, while matrices in sludge samples markedly reduced the effectiveness of PMA treatment. Compared to qPCR, PMA-qPCR results were similar and highly linearly correlated to those obtained by culture assay, indicating that DNA from non-viable cells present in WWTP samples can be eliminated by PMA treatment, and that PMA-qPCR is a reliable method for detection of viable bacteria in environmental samples. This study demonstrated that PMA-qPCR is a rapid and selective detection method for viable bacteria in WWTP samples, and that WWTPs have an obvious function in removing both viable and non-viable bacteria. The results proved that PMA-qPCR is a promising detection method that has a high potential for application as a complementary method to the standard culture-based method in the future.
语种	英语 ; 英语
内容类型	期刊论文
源URL	[http://ir.lib.tsinghua.edu.cn/ir/item.do?handle=123456789/144155]
专题	清华大学
推荐引用方式 GB/T 7714	Dan Li,Tiezheng Tong,Siyu Zeng,et al. Quantification of viable bacteria in wastewater treatment plants by using propidium monoazide combined with quantitative PCR(PMA-qPCR)[J],2016, 2016.
APA	Dan Li.,Tiezheng Tong.,Siyu Zeng.,Yiwen Lin.,Shuxu Wu.,...&Miao He.(2016).Quantification of viable bacteria in wastewater treatment plants by using propidium monoazide combined with quantitative PCR(PMA-qPCR)..
MLA	Dan Li,et al."Quantification of viable bacteria in wastewater treatment plants by using propidium monoazide combined with quantitative PCR(PMA-qPCR)".(2016).

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