Improvement of bovine pestiviral diagnosis by the development of a cost-effective method for detecting viral RNA in fresh specimens and samples spotted in filter papers

IF 1.8 4区生物学 Q4 MICROBIOLOGY Revista Argentina de microbiologia Pub Date : 2023-04-01 DOI:10.1016/j.ram.2022.10.002

Paula M. Favaro , Ana Molineri , Maria J. Dus Santos , Luis F. Calvinho , Andrea Pecora

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Abstract

Bovine pestiviruses are the causative agents of bovine viral diarrhea, a disease that causes severe economic losses in cattle. The aim of this study was to improve their diagnosis by developing a RT-qPCR to detect bovine pestiviruses A, B and H; and to set up a protocol for collecting, shipping and preserving bovine pestiviral RNA on filter papers. The developed RT-qPCR showed high sensitivity in detecting these viruses in different matrices: viral stocks, semen and serum samples. With regard to the possibility of using the technique to test serum pools, it was possible to identify a positive serum sample within a pool containing 30 sera. In addition to evaluating the qPCR from fresh samples, the use of filter papers to sow bovine samples was analyzed. The sampling method on two different filter papers using bovine blood drops was a useful alternative for diagnostic purposes and allowed to preserve pestiviral RNA for up to 12 months under refrigeration.

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通过开发一种具有成本效益的方法来检测新鲜标本和滤纸上发现的样品中的病毒RNA，从而改进牛鼠疫病毒的诊断

牛鼠疫病毒是牛病毒性腹泻的病原体，这种疾病会给牛造成严重的经济损失。本研究的目的是建立RT-qPCR检测牛鼠疫病毒a、B和H;并建立一个收集、运输和保存牛瘟病毒RNA在滤纸上的协议。所建立的RT-qPCR在病毒库、精液和血清样品中检测这些病毒具有较高的灵敏度。关于使用该技术检测血清池的可能性，有可能在含有30份血清的血清池中鉴定出阳性血清样本。除了评估新鲜样品的qPCR外，还分析了使用滤纸播种牛样品的情况。使用牛血滴在两种不同的滤纸上取样的方法是诊断目的的一种有用的替代方法，并且允许在冷藏下保存鼠疫病毒RNA长达12个月。

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来源期刊

Revista Argentina de microbiologia MICROBIOLOGY-

CiteScore

3.30

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： La Revista Argentina de Microbiología es una publicación trimestral editada por la Asociación Argentina de Microbiología y destinada a la difusión de trabajos científicos en las distintas áreas de la Microbiología. La Asociación Argentina de Microbiología se reserva los derechos de propiedad y reproducción del material aceptado y publicado.