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.
Highlights
IntroductionPorcine reproductive and respiratory syndrome is one of the most widespread and economically devastating diseases in the global swine industry [1,2]
PRRS virus (PRRSV) genome was identified as multiAs multifocallydistributed, distributed, individual, individual, or or coalescing coalescing red red dots dots on on the the sections sections prepared prepared from the the focally lungs of the challenged animals
The results prove that PRRSV detection by RNAscope in situ hybridization (ISH) and subsequent digital image analysis can be a powerful tool to assess the viral burden in a histological slide, where the tissue structure is visible
Summary
Porcine reproductive and respiratory syndrome is one of the most widespread and economically devastating diseases in the global swine industry [1,2]. PRRSV is characterized by a high degree of genetic diversity and variability between and within the two species [4,5]. The hemoglobin/haptoglobin scavenger receptor CD163 has been identified as the primary receptor for virus entry into the target cells, as genome-edited animals with a knock-out of either the entire CD163 or just the virus interaction site were resistant to infection [10,11]. Clinical symptoms of PRRS can be very diverse, ranging from asymptomatic infections to outbreaks of high fever and hemorrhagic disease with high morbidity and mortality. Affected piglets and fatteners show respiratory symptoms and, in boars, the semen quality can deteriorate [12,13]
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