Development of Multiplex RT qPCR Assays for Simultaneous Detection and Quantification of Faecal Indicator Bacteria in Bathing Recreational Waters

In this study, we designed and validated in silico and experimentally a rapid, sensitive, and specific multiplex RT qPCR for the detection and quantification of faecal indicator bacteria (FIB) used as microbiological references in marine bathing water regulations ( and intestinal enterococci). The 1...

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Bibliographic Details
Published in:Microorganisms (Basel) 2024-06, Vol.12 (6), p.1223
Main Authors: Carrasco-Acosta, Marina, Garcia-Jimenez, Pilar
Format: Article
Language:English
Subjects:
16S rRNA
Allantoin
Bacteria
Bathing
Climatic changes
Copy number
Design
E coli
Environmental monitoring
Environmental regulations
Escherichia
Escherichia coli
Fecal coliforms
FIB
intestinal enterococci
Intestine
Low concentrations
Marine sediments
multiplex RT qPCR
Multiplexing
Polyethylene
Protein transport
Recreation
Recreational waters
Regulations
Respiratory tract diseases
rRNA 16S
Sea-water
Seawater
Sediments
Shigella
Water quality
Waterborne infections
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Summary:In this study, we designed and validated in silico and experimentally a rapid, sensitive, and specific multiplex RT qPCR for the detection and quantification of faecal indicator bacteria (FIB) used as microbiological references in marine bathing water regulations ( and intestinal enterococci). The 16S rRNA gene was used to quantify group-specific enterococci and and species-specific such as and . Additionally, a gene encoding allantoin transporter protein was used to detect . An assessment of marine coastal systems (i.e., marine water and sediment) revealed that intestinal enterococci were the predominant group compared to / . The low contribution of to the intestinal enterococci group was reported. As and were reported at low concentrations, it is assumed that other enterococci of faecal origin are contributing to the high gene copy number of this group-specific enterococci. Moreover, low 16S rRNA gene copy numbers with respect to and were reported in seawater compared to marine sediment. We conclude that marine sediments can affect the quantification of FIBs included in bathing water regulations. Valuing the quality of the marine coastal system through sediment monitoring is recommended.
ISSN:2076-2607
2076-2607
DOI:10.3390/microorganisms12061223