Epidemiological outbreak investigations were conducted on NADC30-like porcine reproductive and respiratory syndrome virus (PRRSV) to investigate the prevalence of the disease in south-east China in 2015. Two more provinces were found to have NADC30-like PRRSV circulating besides previously reported six provinces. Phylogenetic analysis showed that these virus isolates were clustered in an independent branch and shared high nucleotide similarity to NADC30, a type 2 PRRSV that has been isolated in Unite States in 2008. One NADC30-like PRRSV strain from Henan province was successfully isolated on porcine alveolar macrophages and was tested on 6-week-old specific pathogen-free pigs for pathogenic study. The virus-inoculated pigs showed typical PRRSV clinical symptoms, but all pigs survived throughout the study with a period of 14 days. At necropsy, the lungs of infected pigs developed PRRSV-specific interstitial pneumonia, and virus antigen was detected in lung samples. Therefore, our results indicated NADC30-like PRRSV has widely spread in China and could cause clinical disease on pigs.
Porcine reproductive and respiratory syndrome virus (PRRSV) has varied constantly and circulated in the pig industry worldwide. The prevention and control of porcine reproductive and respiratory syndrome (PRRS) is complicated. A visual, sensitive and specific diagnostic method is advantageous to the control of PRRS. The collateral cleavage activity of LwCas13a is activated to degrade non‐targeted RNA, when crRNA of LwCas13a bond to target RNA. The enhanced Cas13a detection is the combination of collateral cleavage activity of LwCas13a and recombinase polymerase amplification (RPA). In this study, the enhanced Cas13a detection for PRRSV was established. The novel method was an isothermal detection at 37°C, and the detection can be used for real‐time analysis or visual readout. The detection limit of the enhanced Cas13a detection was 172 copies/μl, and there were no cross‐reactions with porcine circovirus 2, porcine parvovirus, classical swine fever virus and pseudorabies virus. The enhanced Cas13a detection can work well in clinical samples. In summary, a visual, sensitive and specific nucleic acid detection method based on CRISPR‐Cas13a was developed for PRRSV.
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