Field-Applicable Recombinase Polymerase Amplification Assay for Rapid Detection of Mycoplasma capricolum subsp. capripneumoniae

摘要: Contagious caprine pleuropneumonia (CCPP) is a highly contagious disease caused by Mycoplasma capricolum subsp. capripneumoniae that affects goats in Africa and Asia. Current available methods for the diagnosis of infection, including cultivation, serological assays, PCR, are time-consuming require fully equipped stationary laboratories, which make them incompatible with testing resource-poor settings most relevant to this disease. We report rapid, specific, sensitive assay employing isothermal DNA amplification using recombinase polymerase (RPA) detection M. capripneumoniae. developed specific target sequence capripneumoniae, as found genome field strain ILRI181 type F38 was further evidenced 10 strains from different geographical regions. Detection limits corresponding 5 × 10(3) 10(4) cells/ml were obtained genomic bacterial culture ILRI181, while no 71 related isolates or Acholeplasma Pasteurella isolates, demonstrating high degree specificity. The produces fluorescent signal within 15 20 min worked well pleural fluid directly CCPP-positive animals without prior extraction. demonstrate CCPP can be achieved, short sample preparation time simple read-out device powered car battery, <45 simulated setting.