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Vol 57(2023) N 3 p. 544-549; DOI 10.1134/S0026893323030068 Full Text

S.A. Lapa1*, S.A. Surzhikov1, S.A. Blagodatskikh2, V.E. Shershov1, A.V. Chudinov1

Recombinase Polymerase Amplification for Rapid Detection of Human Bacterial Pneumonia Pathogens

1Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia
2Scientific Center of Applied Microbiology and Biotechnology, Obolensk, Moscow oblast, 142279 Russia

*lapa@biochip.ru
Received - 2022-11-02; Revised - 2022-11-19; Accepted - 2022-11-20

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 that are similar in size. Identification of the pathogen was carried out by visual analysis of an 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 H37rv, and amounted to 100%. The execution time of the analysis is less than an 1 h, including the electrophoretic reaction control. The test system can be used in specialized clinical laboratories for rapid analysis of samples from patients with suspected pneumonia.

recombinase polymerase amplification, RPA, bacterial pneumonia



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