Comparative analysis of beet western yellows virus detection methods: Development and validation of a novel real-time RT-PCR assay also targeting beet leaf yellowing virus

IF 1.7 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods Pub Date : 2026-05-01 Epub Date: 2026-01-22 DOI:10.1016/j.jviromet.2026.115352

Hellin Pierre , Everaert Ellen , Demonty Elisabeth , Richet Pauline , Muhovski Yordan , De Jonghe Kris , Hautier Louis , Steyer Stéphan

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引用次数: 0

Abstract

Beet western yellows virus (BWYV – Polerovirus BWYV) is a polerovirus associated with yellowing and stunting symptoms in sugar beet and several other crops. Its close relationship with other beet-infecting poleroviruses, has long complicated accurate diagnosis due to serological cross-reactivity and limited molecular data. To overcome these issues, we developed a novel real-time RT-PCR assay enabling the specific and sensitive detection of BWYV and beet leaf yellowing virus (BLYV – Polerovirus BLYV). The assay was designed from a curated genomic dataset to ensure inclusivity and tested in parallel with validated reference methods, including a semi-generic ELISA and a generic polerovirus RT-PCR. Comprehensive validation following the EPPO PM 7/98(5) standard confirmed the method’s high analytical sensitivity, specificity, repeatability, and reproducibility. The assay proved robust to methodological variations and compatible with multiplex diagnostic workflows. Compared with existing approaches, it offers faster, more reliable detection and eliminates cross-reactivity with non-target poleroviruses. This method provides a valuable tool for accurate BWYV surveillance and supports improved management of beet virus yellows in agricultural and phytosanitary contexts.

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甜菜西部黄病毒检测方法的比较分析：一种新的实时RT-PCR检测方法的建立和验证，也针对甜菜叶黄病毒。

甜菜西部黄病毒（BWYV - Polerovirus BWYV）是一种与甜菜和其他几种作物的发黄和发育迟缓症状相关的Polerovirus。它与其他感染甜菜的多极病毒关系密切，由于血清学交叉反应性和有限的分子数据，长期以来使准确诊断复杂化。为了克服这些问题，我们开发了一种新的实时RT-PCR检测BWYV和甜菜叶变黄病毒（BLYV - Polerovirus BLYV）的特异性和敏感性。该检测是根据精心设计的基因组数据集设计的，以确保包容性，并与经过验证的参考方法（包括半通用ELISA和通用多病毒RT-PCR）并行测试。按照EPPO PM 7/98(5)标准进行全面验证，证实该方法具有较高的分析灵敏度、特异性、重复性和再现性。该试验证明了方法变化的稳健性，并与多种诊断工作流程兼容。与现有方法相比，它提供了更快、更可靠的检测，并消除了与非靶标多病毒的交叉反应。该方法为准确监测BWYV提供了有价值的工具，并支持在农业和植物检疫环境下改进甜菜病毒黄的管理。

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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.