Journal
VETERINARY SCIENCES
Volume 8, Issue 10, Pages -Publisher
MDPI
DOI: 10.3390/vetsci8100235
Keywords
porcine reproductive and respiratory syndrome virus; RNAscope; in situ hybridization; qRT-PCR; QuPath
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Funding
- Boehringer Ingelheim Vetmedica GmbH
- [TKP2020-NKA-01]
- [2020-4.1.1-TKP2020]
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A novel RNA-based in situ hybridization technique (RNAscope) was developed to detect PRRSV RNA in lung tissues of experimentally infected animals, showing significant association with real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) test results.
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 R-2 = 0.3894, p = 0.004). This is the first time RNAscope assay has been implemented for the detection of PRRSV-1 in experimental animals.
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