Key result
The MLPA assay demonstrated 100% sensitivity and 99.2% to 100% specificity for detecting 6 porcine reproductive failure-associated viruses compared to real-time PCR.
The MLPA assay provides a fast and accurate screening tool for porcine reproductive failure-associated viruses with comparable sensitivity to real-time PCR.
May enable faster multiplex screening in porcine research; leaves open clinical adoption without prospective validation.
We developed a multiplex ligation-dependent probe amplification (MLPA) assay for the simultaneous detection of 6 clinically relevant viral pathogens causing porcine reproductive failure, that is porcine reproductive and respiratory syndrome virus (PRRSV), Japanese encephalitis virus (JEV), classical swine fever virus (CSFV), porcine circovirus type 2 (PCV2), pseudorabies virus (PRV) and porcine parvovirus (PPV). The limits of detection for the assay varied among the 6 target organisms from 1 to 8 copies per MLPA assay. The MLPA assay was evaluated with 346 heparinized porcine umbilical cord blood specimens, and the results of the assay were compared to those of real-time PCR. The MLPA assay showed specificities and sensitivities of 99.2% and 100%, respectively, for PRRSV; 100% and 100%, respectively, for CSFV, PCV2, PRV and PPV. No sample was found to be positive for JEV by either the MLPA assay or the real-time PCR. In conclusion, the MLPA assay has comparable clinical sensitivity to that of real-time PCR assay and provides a useful tool for fast screening porcine reproductive failure-associated viruses.
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Zhou et al. (2020) studied Porcine reproductive failure-associated viruses (n=346). Multiplex ligation-dependent probe amplification (MLPA) assay vs. Real-time PCR was evaluated on Sensitivity and specificity for detecting 6 viral pathogens. The MLPA assay demonstrated 100% sensitivity and 99.2% to 100% specificity for detecting 6 porcine reproductive failure-associated viruses compared to real-time PCR.
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