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诊断样本中猪病毒RNA靶标的保存策略评估

Assessment of Strategies for Preserving Swine Viral RNA Targets in Diagnostic Specimens.

作者信息

Munguía-Ramírez Berenice, Giménez-Lirola Luis, Zimmerman Jeffrey

机构信息

Veterinary Diagnostic and Production Animal Medicine Department, College of Veterinary Medicine, Iowa State University, Ames, IA 50011, USA.

出版信息

Microorganisms. 2024 Feb 18;12(2):410. doi: 10.3390/microorganisms12020410.

Abstract

Successful downstream molecular analyses of viral ribonucleic acid (RNA) in diagnostic laboratories, e.g., reverse transcription-quantitative polymerase chain reaction (RT-qPCR) or next-generation sequencing, are dependent on the quality of the RNA in the specimen. In swine specimens, preserving the integrity of RNA requires proper sample handling at the time the sample is collected on the farm, during transport, and in the laboratory until RNA extraction is performed. Options for proper handling are limited to maintaining the cold chain or using commercial specimen storage matrices. Herein, we reviewed the refereed literature for evidence that commercial specimen storage matrices can play a role in preserving swine viral RNA in clinical specimens. Refereed publications were included if they compared RNA detection in matrix-treated vs. untreated samples. At present, the small number of refereed studies and the inconsistency in reported results preclude the routine use of commercial specimen storage matrices. For example, specimen storage matrices may be useful under specific circumstances, e.g., where it is mandatory to render the virus inactive. In a broader view, statistically sound side-by-side comparisons between specimens, viral RNA targets, and storage conditions are needed to establish if, when, and how commercial specimen storage matrices could be used in diagnostic medicine.

摘要

在诊断实验室中,对病毒核糖核酸(RNA)进行成功的下游分子分析,例如逆转录定量聚合酶链反应(RT-qPCR)或下一代测序,取决于样本中RNA的质量。在猪样本中,要保持RNA的完整性,需要在农场采集样本时、运输过程中以及实验室中进行RNA提取之前进行适当的样本处理。适当处理的选择仅限于保持冷链或使用商业样本储存基质。在此,我们查阅了相关文献,以寻找商业样本储存基质可在保存临床样本中猪病毒RNA方面发挥作用的证据。如果参考文献比较了经基质处理和未经处理的样本中的RNA检测情况,则将其纳入。目前,由于参考文献研究数量较少且报告结果不一致,商业样本储存基质无法常规使用。例如,样本储存基质可能在特定情况下有用,例如在必须使病毒失活的情况下。从更广泛的角度来看,需要对样本、病毒RNA靶点和储存条件进行统计学上合理的并列比较,以确定商业样本储存基质是否、何时以及如何用于诊断医学。

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