6-plex Crystal Digital PCR® 用于全面监测废水中的呼吸道和食源性细菌病原体

IF 7.6 2区环境科学与生态学 Q1 ENVIRONMENTAL SCIENCES

Environmental Pollution Pub Date : 2025-04-22 DOI:10.1016/j.envpol.2025.126298

Sadhana Shrestha, Bikash Malla, Eiji Haramoto

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引用次数: 0

摘要

尽管世界范围内细菌性呼吸道和胃肠道感染的负担很高，但与病毒性WS相比，细菌性废水监测（WS）是一个较少探索的领域。本研究建立了6-plex Crystal Digital PCR®（cdPCR）系统，对废水中的急性呼吸道病原菌a族化脓性链球菌、食源性疾病病原菌产气荚膜梭菌、沙门氏菌、空肠弯曲杆菌和大肠杆菌以及指示菌大肠杆菌进行综合监测。在2023年6月至2024年5月期间，每周从日本山梨县的一家污水处理厂收集52份抓取液样本进行离心，然后进行DNA提取和cdPCR。采用naica®系统（Stilla Technologies）进行cdPCR。6-plex cdPCR检测显示了较强的性能。52份样品中产气荚膜荚膜菌阳性率为100%，沙门氏菌阳性率为98%，空肠菌阳性率为56%，大肠杆菌阳性率为25%，化脓性链球菌阳性率为63%，浓度范围为4.2±0.3 ~ 7.5±0.2 log10 copies/L。产气荚膜荚膜梭菌浓度显著高于其他病原菌(单因素方差分析，p值<；0.05)，表明其优势性。沙门氏菌检出率较高，表明人群中沙门氏菌感染率较高。除大肠杆菌检出率外，其他病原菌均无季节变化。春季化脓性链球菌的浓度明显高于其他季节，与该流域化脓性链球菌咽炎病例的趋势一致。总之，6-plex cdPCR系统是一种有价值的综合WS工具，对公共卫生监测具有重要意义。

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6-plex Crystal Digital PCR® for comprehensive surveillance of respiratory and foodborne bacterial pathogens in wastewater

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6-plex Crystal Digital PCR® for comprehensive surveillance of respiratory and foodborne bacterial pathogens in wastewater

Bacterial wastewater surveillance (WS) is less explored area compared to viral WS despite high burden of bacterial respiratory and gastrointestinal infections worldwide. This study established a 6-plex Crystal Digital PCR® (cdPCR) system, to comprehensively monitor an acute respiratory pathogen - Group A Streptococcus (GAS) pyogenes, foodborne disease (FBD) pathogens - Clostridium perfringens, Salmonella spp., Campylobacter jejuni, and Campylobacter coli, and an indicator bacterium, Escherichia coli in wastewater. Fifty-two grab influent samples collected weekly from a wastewater treatment plant in Yamanashi Prefecture, Japan, between June 2023 and May 2024 were centrifuged, followed by DNA extraction and cdPCR. cdPCR was performed using the naica® system (Stilla Technologies). The 6-plex cdPCR assays showed strong performance. Among the 52 samples, 100 % of samples were positive for C. perfringens, 98 % for Salmonella spp., 56 % for C. jejuni, 25 % for C. coli, and 63 % for S. pyogenes, with concentrations ranging between 4.2 ± 0.3 to 7.5 ± 0.2 log₁₀ copies/L. The concentration of C. perfringens was significantly higher than that of other pathogens (p < 0.05), indicating its dominance. Salmonella spp. had high detection rate, implying increased Salmonella infection in the population. Seasonal variation was not observed in any of FBD pathogens, except for the detection rate of C. coli. S. pyogenes concentrations were significantly higher in spring than in other seasons, agreeing with the trend of GAS pharyngitis cases in the catchment. In conclusion, the 6-plex cdPCR system is a valuable tool for comprehensive WS, offering significant implications for public health monitoring.

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来源期刊

Environmental Pollution 环境科学-环境科学

CiteScore

16.00

自引率

6.70%

发文量

2082

审稿时长

2.9 months

期刊介绍： Environmental Pollution is an international peer-reviewed journal that publishes high-quality research papers and review articles covering all aspects of environmental pollution and its impacts on ecosystems and human health. Subject areas include, but are not limited to: • Sources and occurrences of pollutants that are clearly defined and measured in environmental compartments, food and food-related items, and human bodies; • Interlinks between contaminant exposure and biological, ecological, and human health effects, including those of climate change; • Contaminants of emerging concerns (including but not limited to antibiotic resistant microorganisms or genes, microplastics/nanoplastics, electronic wastes, light, and noise) and/or their biological, ecological, or human health effects; • Laboratory and field studies on the remediation/mitigation of environmental pollution via new techniques and with clear links to biological, ecological, or human health effects; • Modeling of pollution processes, patterns, or trends that is of clear environmental and/or human health interest; • New techniques that measure and examine environmental occurrences, transport, behavior, and effects of pollutants within the environment or the laboratory, provided that they can be clearly used to address problems within regional or global environmental compartments.