A Double Lysis Method for Animal Rotavirus RNA Extraction From Stool Samples

Current protocols Pub Date : 2024-10-04 DOI:10.1002/cpz1.70011

Maame Ekua Acquah, Samuel Mawuli Adadey, Sylvester Languon, Osbourne Quaye

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Abstract

Globally, porcine rotavirus is a leading cause of gastroenteritis in nursing and post-weaning piglets, as well as adult pigs. Between February 2015 and June 2016, 156 fecal samples were collected from pigs in the Northeastern part of Accra, Ghana, and screened for Group A rotavirus using the Proflow^TM Kit. Here, we describe different extraction methods that were employed to recover high-quality RNA for downstream analysis, with emphasis on a novel hybrid extraction method. The hybrid approach with a kit and manual extraction method led to a 10-fold greater RNA yield versus the kit-based method alone. The new extraction method gave an average purity ratio (A₂₆₀/A₂₈₀) of 1.8, which was also significantly higher than that obtained solely from the manual or kit-based extraction methods. Our novel hybrid approach will be useful in the extraction of rotavirus from animal fecal samples, thus improving the yield of RNA for downstream analysis. © 2024 Wiley Periodicals LLC.

Basic Protocol: Hybrid 2: A double lysis method for RNA extraction from animal stool samples

Support Protocol 1: The GenElute extraction method

Support Protocol 2: Hybrid 1 extraction method

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从粪便样本中提取动物轮状病毒 RNA 的双重裂解法。

在全球范围内，猪轮状病毒是哺乳仔猪、断奶后仔猪以及成年猪肠胃炎的主要病因。2015 年 2 月至 2016 年 6 月期间，我们在加纳阿克拉东北部收集了 156 份猪的粪便样本，并使用 ProflowTM 套件进行了 A 组轮状病毒筛查。在此，我们介绍了用于回收高质量 RNA 以进行下游分析的不同提取方法，并重点介绍了一种新型混合提取方法。与单独使用试剂盒提取方法相比，混合使用试剂盒和人工提取方法的RNA产量提高了10倍。新提取方法的平均纯度比（A260/A280）为 1.8，也明显高于单纯的人工或试剂盒提取方法。我们的新型混合方法将有助于从动物粪便样本中提取轮状病毒，从而提高下游分析的 RNA 产量。© 2024 Wiley Periodicals LLC.基本方案：混合 2：从动物粪便样本中提取 RNA 的双重裂解法支持协议 1：GenElute 提取法支持协议 2：混合 1 提取法。

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

Current protocols

CiteScore

4.00

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0.00%

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