Development of a High-Throughput QPCR Assay for Detecting Waterborne Protozoa and Helminths Across Different Environmental Media in China

Overview

Journal China CDC Wkly

Date 2025 Jan 27

PMID 39867816

Authors

Fuming Duan

Ziming Han

Tiantian Tian

Huican Zhang

Min Yang

Yu Zhang

Affiliations

Soon will be listed here.

Abstract

Introduction: The establishment of a high-throughput quantification approach for waterborne pathogenic protozoa and helminths is crucial for rapid screening and health risk assessment.

Methods: We developed a high-throughput quantitative polymerase chain reaction (HT-qPCR) assay targeting 19 waterborne protozoa and 3 waterborne helminths and validated its sensitivity, specificity, and repeatability. The assay was then applied to test various environmental media samples.

Results: The HT-qPCR assay's limit of detection (LOD) was 5×10 copies/μL DNA, and its specificity was confirmed using and standards. Repeatability, assessed through intra- and inter-group experiments, yielded a coefficient of variation () of 1.0%-4.6% and 1.2%-6.4% at concentrations of 1×10 and 1×10 copies/μL, respectively. The values of the 22 standard curves ranged from 0.983 to 0.998, with amplification efficiencies between 80% and 107%. In drinking water sources, sludge from municipal wastewater treatment plants (MWTPs), and livestock manure samples, 17 of 22 targets were detected, with genus (50.0%), (11.8%), and (11.8%) showing high prevalence. spp., , and were simultaneously found in all three sample types.

Discussion: This study presents a useful tool for the rapid detection of waterborne protozoa and helminths in complex environmental microbiomes, providing scientific data for monitoring cross-media transmission and controlling microbial risk from a One Health perspective.

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