Key result
The PCV2 real-time RPA and RPA LFD assays detected PCV2 with a sensitivity of 10^2 copies per reaction within 20 minutes at 37°C, showing high specificity with no cross-reactions.
The developed RPA assays provide a rapid, sensitive, and specific alternative for the detection of Porcine Circovirus Type 2.
RPA assays enable rapid PCV2 detection in labs; leaves open field validation before veterinary adoption.
Porcine circovirus virus type II (PCV2) is the etiology of postweaning multisystemic wasting syndrome (PMWS), porcine dermatitis, nephropathy syndrome (PDNS), and necrotizing pneumonia. Rapid diagnosis tool for detection of PCV2 plays an important role in the disease control and eradication program. Recombinase polymerase amplification (RPA) assays using a real-time fluorescent detection (PCV2 real-time RPA assay) and RPA combined with lateral flow dipstick (PCV2 RPA LFD assay) were developed targeting the PCV2 ORF2 gene. The results showed that the sensitivity of the PCV2 real-time RPA assay was 102copies per reaction within 20 min at 37°C and the PCV2 RPA LFD assay had a detection limit of 102copies per reaction in less than 20 min at 37°C. Both assays were highly specific for PCV2, with no cross-reactions with porcine circovirus virus type 1, foot-and-mouth disease virus, pseudorabies virus, porcine parvovirus, porcine reproductive and respiratory syndrome virus, and classical swine fever virus. Therefore, the RPA assays provide a novel alternative for simple, sensitive, and specific identification of PCV2.
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Yang et al. (2017) studied Porcine circovirus virus type II (PCV2). Recombinase polymerase amplification (RPA) assays was evaluated on Sensitivity and specificity of PCV2 real-time RPA and RPA LFD assays. The PCV2 real-time RPA and RPA LFD assays detected PCV2 with a sensitivity of 10^2 copies per reaction within 20 minutes at 37°C, showing high specificity with no cross-reactions.
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