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Selective Detection of Viable Pathogens in Water Using Propidium Monoazide Combined with qPCR and Its Application

Author: ZuoTieZuo
Tutor: HeMiao
School: Tsinghua University
Course: Environmental Science and Engineering
Keywords: PMA-qPCR Active bacteria Pathogen detection Sewage treatment Disinfection characteristics
CLC: R123.1
Type: Master's thesis
Year: 2010
Downloads: 113
Quote: 0
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Rapid and accurate detection of pathogens in water activity on the prevention of water-borne disease outbreaks and protect water quality and safety is important. In this study, the conventional techniques can not distinguish between active bacteria qPCR and the lack of non-active bacteria, the nucleic acid dye established a PMA (propidium monoazide) combined with qPCR detection method (PMA-qPCR), the full activity of the membrane can be pathogens selective detection, studies using this method to analyze typical pathogen indicator bacteria in the sewage treatment process to remove features, especially for common disinfection disinfection properties of E. coli do in depth. The main conclusions are: (1) PMA dye in qPCR analysis to effectively remove bacteria from non-active DNA, and the DNA not active bacteria significant impact on the detection result, PMA-qPCR method can complete the sample cell activity for selective detection of bacteria, but the method for different types of active bacteria selective detection results vary. (2) PMA-qPCR method is applicable to sewage treatment plant influent, secondary effluent, secondary clarifier sludge activity in real samples of different selective detection of bacteria, but the primary sludge will significantly inhibited PMA-qPCR choice detection results, the application of this method has limitations. (3) PMA-qPCR method of sewage treatment process pathogen indicator bacteria culture test results and test results had a good linear correlation, and the difference was not significant, compared with the conventional qPCR, the method of detection results more close to the culture, indicating that PMA-qPCR method can meet the sewage treatment process active pathogen testing requirements. (4) PMA-qPCR, qPCR and conventional culture method test results together indicate that a treatment process for E. coli and Enterococcus faecalis were no significant removal, secondary treatment process of these two pathogenic indicator bacteria removal In 1.5 log10 or so. (5) PMA-qPCR method to characterize two kinds of chlorine and chloramine disinfection an inactivation of E. coli. The method of chlorine and monochloramine inactivation curves of E. coli meet a kinetic equation, inactivation rate constants were 2.24 L / (mg · min) and 0.0175 L / (mg · min), lower than that for France inactivation rate constants obtained, indicating that the pathogen chlorine and chloramines a rate lower than its destruction of cell membranes may develop resistance to the pathogen destruction rate. However, current research indicates that PMA-qPCR is still unable to accurately assess the UV disinfection for the inactivation of E. coli.

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CLC: > Medicine, health > Preventive Medicine,Health > Environmental health,environmental medicine > Water and water supply sanitation > Health surveillance of the health standards of water and water
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