In Situ Hybridization of PRRSV-1 Combined with Digital Image Analysis in Lung Tissues of Pigs Challenged with PRRSV-1

Abstract

Betaarterivirus suid 1 and 2 are the causative agents of porcine reproductive and respiratory syndrome (PRRS), which is one of the most significant diseases of the swine industry, causing significant economic losses in the main pig producing countries. Here, we report the development of a novel, RNA-based in situ hybridization technique (RNAscope) to detect PRRS virus (PRRSV) RNA in lung tissues of experimentally infected animals. The technique was applied to lung tissues of 20 piglets, which had been inoculated with a wild-type, highly pathogenic PRRSV-1 strain. To determine the RNAscope’s applicability as a semi-quantitative method, we analysed the association between the proportion of the virus-infected cells measured with an image analysis software (QuPath) and the outcome of the real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) tests performed in parallel. The results of the quantitative approach of these two molecular biological methods show significant association (pseudo R2 = 0.3894, p = 0.004). This is the first time RNAscope assay has been implemented for the detection of PRRSV-1 in experimental animals.

Relationship between Uveal Inflammation and Viral Detection in 30 Cats with Feline Infectious Peritonitis

“…Here, while in high agreement with IHC and RT-qPCR, it has proven to have the highest detection rate among the three diagnostic techniques used (10/30). The sensitivity and specificity of this technology has been well described and validated for other pathogens [36][37][38][39][40][41][42][43][44][45] and therefore could be a promising tool for the diagnosis of FIP in tissue sections. Further evaluation of this method in extraocular tissues from FIP-infected cats is warranted to determine its performance.…”

Glycyrrhizic Acid-Loaded Poly-<i>ɛ</i>-Caprolactone Nanoparticles Decrease PRRSV Infection in MARC-145 Cells

“…Porcine reproductive and respiratory syndrome (PRRS) is a viral disease that af-fects domestic pigs of all ages, causing significant economic losses in the swine industry due to its devastating effect on production [1] [2]. The causative agent Betaarterivirus suid (PRRSV) shows great genetic and antigenic variability with a marked increase in virulence, so vaccines should be useful in controlling the problems caused [3] [4] [5].…”