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Recombinase Polymerase Amplification for Rapid Detection of Human Bacterial Pneumonia Pathogens

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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 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.

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Funding

The study was supported by a grant Russian Science Foundation (no. 22-14-00257).

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Authors and Affiliations

  1. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991, Moscow, Russia

    S. A. Lapa, S. A. Surzhikov, V. E. Shershov & A. V. Chudinov

  2. Scientific Center of Applied Microbiology and Biotechnology, 142279, Obolensk, Moscow oblast, Russia

    S. A. Blagodatskikh

Authors
  1. S. A. Lapa
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  2. S. A. Surzhikov
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  3. S. A. Blagodatskikh
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  4. V. E. Shershov
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  5. A. V. Chudinov
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Correspondence to S. A. Lapa.

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The authors declare that they have no conflicts of interest. This article does not contain a description of any research involving humans or animals as subjects.

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Lapa, S.A., Surzhikov, S.A., Blagodatskikh, S.A. et al. Recombinase Polymerase Amplification for Rapid Detection of Human Bacterial Pneumonia Pathogens. Mol Biol 57, 544–549 (2023). https://doi.org/10.1134/S0026893323030068

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  • DOI: https://doi.org/10.1134/S0026893323030068

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