title: Quantification of viable bacteria in wastewater treatment plants by using propidium monoazide combined with quantitative PCR (PMA-qPCR).
abstract: 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 micromol/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.
authors: Dan
authors: D
authors: Li
authors: Tiezheng
authors: T
authors: Tong
authors: Siyu
authors: S
authors: Zeng
authors: Yiwen
authors: Y
authors: Lin
authors: Shuxu
authors: S
authors: Wu
authors: Miao
authors: M
authors: He
mesh_terms: D001386:Azides; D016983:Enterococcus; D004926:Escherichia coli; D016133:Polymerase Chain Reaction; D011419:Propidium / Q000031:analogs & derivatives*; D012722:Sewage / Q000382:microbiology*; D062065:Wastewater / Q000382:microbiology*; D014871:Water Microbiology
chemical_list: D001386:Azides; D011419:Propidium; D012722:Sewage; D062065:Wastewater; C533957:propidium monoazide