Isolation of porcine reproductive and respiratory syndrome virus from feed ingredients and complete feed, with subsequent RT-qPCR analysis

从饲料原料和全价饲料中分离猪繁殖与呼吸综合征病毒,并进行后续的RT-qPCR分析

阅读:1

Abstract

We used virus isolation (VI) to determine tissue culture infectivity and reverse-transcription quantitative PCR (RT-qPCR) to determine the stability of porcine reproductive and respiratory syndrome virus 2 (PRRSV) strain P129 in solvent-extracted soybean meal (SBM), dried distillers grains with solubles (DDGS), complete swine feed (FEED), or medium (DMEM) at 4°C, 23°C, or 37°C for up to 3 d. Samples of each treatment were taken at regular intervals and processed. Supernatant was titrated and used to inoculate confluent MARC-145 cells to determine infectivity. RNA was extracted from each supernatant sample and tested by RT-qPCR to determine any change in detectable virus RNA across matrix type, temperature, and time. An interaction (p = 0.028) was observed for matrix × temperature × hour for live virus detected by VI. At 4°C, the concentration of infectious virus was greatest in DMEM, intermediate in SBM, and lowest in DDGS and FEED. DMEM also had the greatest concentration of infectious PRRSV at 23°C over time; a higher infectious virus concentration was maintained in SBM for longer than in DDGS or FEED. At 37°C, a greater concentration of infectious virus was sustained in DMEM than in the feedstuffs, with concentrations decreasing until 48 h post-inoculation. Only matrix type influenced the quantity of viral RNA detected by RT-qPCR (p = 0.032). More viral RNA was detected in the virus control than in DDGS; SBM and FEED were intermediate. By VI, we found that infectious virus could be harbored in SBM, DDGS, and FEED for a short time.

特别声明

1、本页面内容包含部分的内容是基于公开信息的合理引用;引用内容仅为补充信息,不代表本站立场。

2、若认为本页面引用内容涉及侵权,请及时与本站联系,我们将第一时间处理。

3、其他媒体/个人如需使用本页面原创内容,需注明“来源:[生知库]”并获得授权;使用引用内容的,需自行联系原作者获得许可。

4、投稿及合作请联系:info@biocloudy.com。