Simultaneous detection of various pathogenic Escherichia coli in water by sequencing multiplex PCR amplicons

Waterborne diseases due to pathogen contamination in water are a serious problem all over the world. Accurate and simultaneous detection of pathogens in water is important to protect public health. In this study, we developed a method to simultaneously detect various pathogenic Escherichia coli by s...

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Bibliographic Details
Published in:	Environmental monitoring and assessment Vol. 195; no. 2; p. 264
Main Authors:	Suzuki, Yoshihiro, Shimizu, Hiroki, Tamai, Shouichiro, Hoshiko, Yuki, Maeda, Toshinari, Nukazawa, Kei, Iguchi, Atsushi, Masago, Yoshifumi, Ishii, Satoshi
Format:	Journal Article
Language:	English
Published:	Cham Springer International Publishing 01-02-2023 Springer Nature B.V
Subjects:	Atmospheric Protection/Air Quality Control/Air Pollution Contamination Detection E coli Earth and Environmental Science Ecology Ecotoxicology Environment Environmental Management Environmental monitoring Environmental Monitoring - methods Environmental science Escherichia coli Escherichia coli - genetics Escherichia coli - pathogenicity Genes LT gene Monitoring/Environmental Analysis Multiplex Polymerase Chain Reaction - methods Multiplexing Nucleotide sequence Pathogens PCR Public health River water Rivers Sequence analysis Sequencing Sth gene Wastewater Water Microbiology Water pollution Water sampling Waterborne diseases Sequencing analysis Detection and determination Read number Pathogenic genes Multiplex PCR amplification
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Summary:	Waterborne diseases due to pathogen contamination in water are a serious problem all over the world. Accurate and simultaneous detection of pathogens in water is important to protect public health. In this study, we developed a method to simultaneously detect various pathogenic Escherichia coli by sequencing the amplicons of multiplex PCR. Our newly designed multiplex PCR amplified five genes for pathogenic E. coli ( uidA , stx1 , stx2 , STh gene, and LT gene). Additional two PCR assays (for aggR and eae ) were also designed and included in the amplicon sequencing analysis. The same assays were also used for digital PCR (dPCR). Strong positive correlations were observed between the sequence read count and the dPCR results for most of the genes targeted, suggesting that our multiplex PCR-amplicon sequencing approach could provide quantitative information. The method was also successfully applied to monitor the level of pathogenic E. coli in river water and wastewater samples. The approach shown here could be expanded by targeting genes for other pathogens.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0167-6369 1573-2959
DOI:	10.1007/s10661-022-10863-6