Recombinase Polymerase Amplification for Rapid Detection of Human Bacterial Pneumonia Pathogens

Abstract

A diagnostic system based on recombinase polymerase amplification (RPA) has been developed to identify six bacterial pathogens of human pneumonia. Species-specific primers have been designed and optimized to conduct a multiplex reaction in one common volume. Labeled primers were used for reliable discrimination of amplification products close in size. Identification of the pathogen was carried out by visual analysis of the electrophoregram. The analytical sensitivity of the developed multiplex RPA was 102‒103 copies of DNA. The specificity of the system was determined by the absence of cross-amplification of the studied DNA samples of pneumonia pathogens for each pair of primers, as well as for the DNA of Mycobacterium tuberculosis H37rν, and amounted to 100%. The analysis execution time is less than an 1 h, including electrophoretic reaction control. The test system can be used in specialized clinical laboratories for rapid analysis of samples from patients with suspected pneumonia.