Background: Canine distemper, caused by Canine distemper virus (CDV), is a highly contagious and fatal systemic\ndisease in free-living and captive carnivores worldwide. Recombinase polymerase amplification (RPA), as an\nisothermal gene amplification technique, has been explored for the molecular detection of diverse pathogens.\nMethods: A real-time reverse transcription RPA (RT-RPA) assay for the detection of canine distemper virus\n(CDV) using primers and exo probe targeting the CDV nucleocapsid protein gene was developed. A series of\nother viruses were tested by the RT-RPA.Thirty-two field samples were further tested by RT-RPA, and the\nresuts were compared with those obtained by the real-time RT-PCR.\nResults: The RT-RPA assay was performed successfully at 40 Ã?°C, and the results were obtained within 3 minââ?¬â??\n12 min. The assay could detect CDV, but did not show cross-detection of canine parvovirus-2 (CPV-2), canine\ncoronavirus (CCoV), canine parainfluenza virus (CPIV), pseudorabies virus (PRV) or Newcastle disease virus\n(NDV), demonstrating high specificity. The analytical sensitivity of RT-RPA was 31.8 copies in vitro transcribed\nCDV RNA, which is 10 times lower than the real-time RT-PCR. The assay performance was validated by testing 32\nfield samples and compared to real-time RT-PCR. The results indicated an excellent correlation between RT-RPA\nand a reference real-time RT-PCR method. Both assays provided the same results, and R2 value of the positive\nresults was 0.947.\nConclusions: The results demonstrated that the RT-RPA assay offers an alternative tool for simple, rapid, and\nreliable detection of CDV both in the laboratory and point-of-care facility, especially in the resource-limited\nsettings
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