Simultaneous detection of various pathogenic Escherichia coli in water by sequencing multiplex PCR amplicons.
Environ Monit Assess
; 195(2): 264, 2023 Jan 05.
Article
em En
| MEDLINE
| ID: mdl-36600083
ABSTRACT
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.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Contexto em Saúde:
2_ODS3
/
3_ND
Base de dados:
MEDLINE
Assunto principal:
Microbiologia da Água
/
Escherichia coli
/
Reação em Cadeia da Polimerase Multiplex
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Revista:
Environ Monit Assess
Ano de publicação:
2023
Tipo de documento:
Article