Comparative Analysis of RT-qPCR Assay Sensitivity and Process Limit of Detection for Japanese Encephalitis Virus (JEV) Detection in Piggery Wastewater.

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods Pub Date : 2025-10-01 DOI:10.1016/j.jviromet.2025.115272

Angela Freddi, Metasebia Gebrewold, Wendy J M Smith, Manini Elana, Stuart L Simpson, Warish Ahmed

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引用次数: 0

Abstract

Japanese encephalitis virus (JEV) poses a significant public health threat in Asia, the Western Pacific, and Australia, necessitating robust surveillance and management strategies. This study evaluates three RT-qPCR assays (Universal JEV, JEV G4, and VIDRL2 JEV G4) for detecting JEV in piggery wastewater, a promising approach for early outbreak detection. We assessed assay limit of detection (ALOD), process limit of detection (PLOD), and recovery efficiency using gamma-irradiated JEV seeded into wastewater samples, alongside field-derived samples from an Australian piggery. The JEV G4 assay demonstrated superior sensitivity, with an ALOD of 2.2 to 5.7 copies/reaction and PLOD of 72 to 282 copies/10mL of piggery wastewater, detecting JEV in 24/30 field samples compared to 17/30 for Universal JEV and 0/30 for VIDRL2 JEV G4. Recovery efficiencies varied, with JEV G4 showing consistent performance (14.9-26.6%) across concentrations. McNemar's test confirmed JEV G4's higher sensitivity (p < 0.05). Based on the results obtained in this study, the JEV G4 assay is recommended for wastewater surveillance in genotype 4 regions, with a dual-assay approach suggested for broader genotype coverage. These findings enhance JEV surveillance strategies, supporting early detection and control.

查看原文本刊更多论文

猪舍废水中流行性乙型脑炎病毒（JEV） RT-qPCR检测灵敏度及工艺限的比较分析

日本脑炎病毒（JEV）在亚洲、西太平洋和澳大利亚构成重大公共卫生威胁，需要强有力的监测和管理战略。本研究评估了三种RT-qPCR方法（通用乙脑病毒、乙脑病毒G4和VIDRL2乙脑病毒G4）检测猪场废水中的乙脑病毒，这是一种很有前景的早期爆发检测方法。我们利用γ辐照的乙脑病毒注入废水样品，以及从澳大利亚养猪场提取的样品，评估了检测限（ALOD）、过程检测限（PLOD）和回收率。JEV G4检测显示出卓越的灵敏度，ALOD为2.2 ~ 5.7拷贝/反应，PLOD为72 ~ 282拷贝/10mL猪场废水，在24/30的现场样品中检测到JEV，而Universal JEV为17/30，VIDRL2 JEV G4为0/30。在不同浓度下，乙脑病毒G4的回收率保持一致（14.9 ~ 26.6%）。McNemar试验证实乙脑病毒G4具有较高的敏感性（p < 0.05）。基于本研究的结果，建议采用乙脑病毒G4检测方法对基因型4区域的废水进行监测，建议采用双检测方法以扩大基因型覆盖范围。这些发现加强了乙脑病毒监测战略，支持早期发现和控制。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of virological methods 医学-病毒学

CiteScore

5.80

自引率

0.00%

发文量

209

审稿时长

41 days

期刊介绍： The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.