Development and evaluation of a colorimetric LAMP based-assay targeting the Bacteroides HF183 marker for tracking sewage pollution in environmental waters.

Autor: do Nascimento MCA; CSIRO Environment, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia; Department of Biology, São Paulo State University - UNESP, São José do Rio Preto, São Paulo 15054-000, Brazil., Smith WJM; CSIRO Environment, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia., Gebrewold M; CSIRO Environment, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia., Liu Y; CSIRO Environment, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia; State Key Laboratory of Marine Environmental Science, College of the Environment & Ecology, Xiamen University, Xiamen 361102, China., Simpson SL; CSIRO Environment, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia., Bivins A; Department of Civil & Environmental Engineering, Louisiana State University, Baton Rouge, LA 70803, USA., Rahal P; Department of Biology, São Paulo State University - UNESP, São José do Rio Preto, São Paulo 15054-000, Brazil., Ahmed W; CSIRO Environment, Ecosciences Precinct, 41 Boggo Road, QLD 4102, Australia. Electronic address: Warish.Ahmed@csiro.au.
Jazyk: angličtina
Zdroj: Water research [Water Res] 2024 Oct 15; Vol. 264, pp. 122202. Date of Electronic Publication: 2024 Jul 31.
DOI: 10.1016/j.watres.2024.122202
Abstrakt: Surface waters are vulnerable to contamination by human and animal feces, posing risks to human health due to potential exposure to enteric pathogens. This research developed a colorimetric loop-mediated isothermal amplification (cLAMP) assay to detect sewage associated Bacteroides dorei HF183/BacR287 (HF183) marker in wastewater and environmental water samples. The host sensitivity and host specificity of the assay were evaluated, and their performance was compared to the Bacteroides HF183 qPCR assay using control materials (gBlocks), environmental water samples seeded with untreated sewage, and ambient environmental water samples. In serial dilutions of control materials, qPCR produced quantifiable data across all dilutions, while cLAMP detected the marker down to 0.001 pg/µL of control materials, which was two orders of magnitude less sensitive than qPCR. All untreated sewage samples (n = 12) tested positive for HF183 by both the qPCR and cLAMP assays, demonstrating a host sensitivity value of 1.00 (maximum value of 1.00). The host specificity by analysing 70 non-human fecal nucleic acid samples revealed cLAMP's specificity value of 0.81 compared to qPCR's 0.64. When testing sewage-seeded environmental water samples, both methods detected HF183 for the lowest amount of sewage, indicating similar detection sensitivity. The application of cLAMP for tracking sewage pollution in environmental waters showed promising results, with moderate agreement between cLAMP and qPCR (κ = 0.510). However, cLAMP occasionally missed detections compared to qPCR, particularly in low-concentration samples. Overall, the cLAMP HF183 assay demonstrated promising potential as a rapid and sensitive method for detecting sewage pollution, offering a viable alternative to qPCR in certain environmental monitoring scenarios.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)
Databáze: MEDLINE
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