有效的RNA提取与现成的试剂：替代方案的比较分析

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods Pub Date : 2025-08-19 DOI:10.1016/j.jviromet.2025.115242

Sandra González G , Ana Dias , Rodrigo Reis , Shirley Villalba , Cynthia Vazquez , Marilda Mendonça Siqueira , Fernando do Couto Motta

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引用次数: 0

摘要

SARS-CoV-2大流行暴露了诊断实验室工作流程方面的重大挑战，试剂和自动化的短缺加剧了对检测需求激增的管理。本研究评估了四种直接样品制备方案，并提出了一种经济的替代RNA提取方法，使用通用硅柱作为资源短缺的可行解决方案。虽然测试的非提取方案产生假阴性的风险较高，但替代方法显示的结果在统计上与标准手动和自动提取方案相当（方差分析，F=0.69, p = 0.415），平均Ct差异仅为一个周期。该方案表明，只有在极端病毒载量下效率才会降低，并为在日益普遍的流行病压力下面临供应链限制的公共卫生实验室提供了一种具有成本效益和可获得的选择。

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Effective RNA extraction with readily available reagents: A comparative analysis of alternative protocols

The SARS-CoV-2 pandemic exposed significant challenges in diagnostic laboratory workflows, where shortages of reagents and automation worsened the management of surges in testing demand. This study evaluates four direct sample preparation protocols and proposes an economical alternative RNA extraction method using generic silica columns as a feasible solution during resource scarcity. While the non-extraction protocols tested presented an elevated risk of producing false negatives, the alternative method demonstrated results statistically comparable to standard manual and automated extraction protocols (ANOVA, F=0.69, p = 0.415), with an average Ct difference of only one cycle. The protocol demonstrated reduced efficiency only at extreme viral loads and offers a cost-effective, accessible option for public health laboratories facing supply chain constraints during increasingly common epidemic pressures.

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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.