Effect of extrinsic factors on the detection of PRRSV and a porcine-specific internal sample control in serum, oral fluid, and fecal specimens tested by RT-rtPCR

Author:

Munguía-Ramírez Berenice1ORCID,Armenta-Leyva Betsy1ORCID,Henao-Díaz Alexandra2,Cheng Ting-Yu3ORCID,Zhang Jianqiang1,Rawal Gaurav1,Ye Fangshu4ORCID,Giménez-Lirola Luis1ORCID,Zimmerman Jeffrey J.1ORCID

Affiliation:

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

2. Pig Improvement Company México, Santiago de Querétaro, Querétaro, México

3. Department of Veterinary Preventive Medicine, The Ohio State University, Columbus, OH, USA

4. Department of Statistics, College of Liberal Arts and Sciences, Iowa State University, Ames, IA, USA

Abstract

We characterized the effect of 1) temperature × time, 2) freeze-thaw cycles, and 3) high porcine reproductive and respiratory syndrome virus (PRRSV) RNA concentrations on the detection of PRRSV and a porcine-specific internal sample control (ISC) in serum, oral fluid, and fecal samples using a commercial PRRSV RT-rtPCR assay (Idexx). In study 1, the effect of temperature × time on PRRSV and ISC detection was shown to be specimen dependent. In serum stored at 4, 10, or 20°C, PRRSV detection was consistent for up to 168 h, but storage at 30°C reduced detectable PRRSV RNA. ISC RNA was stable in serum held at 4 and 10°C, but not at 20 and 30°C. In contrast, PRRSV and ISC RNAs in oral fluid and fecal samples continuously decreased at all temperature × time treatments. Based on these data, serum samples should be stored at ≤ 20°C to optimize PRRSV RNA detection. Oral fluid and fecal samples should be frozen in a non–self-defrosting freezer until tested. In study 2, freeze-thaw cycles had little impact on PRRSV and ISC detection, but more so in oral fluids than serum or fecal samples. Thus, freeze-thaw cycles in oral fluids should be minimized before RT-rtPCR testing. In study 3, the ISC was not affected by high concentrations of PRRSV RNA in serum, oral fluid, or fecal samples. It should not be assumed that data from our PRRSV study are applicable to other pathogens; additional pathogen-specific studies are required.

Publisher

SAGE Publications

Subject

General Veterinary

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