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 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 $ 10^{2} $–$ 10^{3} $ 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 $ H37_{rv} $, 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..
| Medienart: |
Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:57 |
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| Enthalten in: |
Molecular biology - 57(2023), 3 vom: Juni, Seite 544-549 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Lapa, S. A. [VerfasserIn] |
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| Links: |
Volltext [lizenzpflichtig] |
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| BKL: |
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| Anmerkungen: |
© Pleiades Publishing, Inc. 2023. ISSN 0026-8933, Molecular Biology, 2023, Vol. 57, No. 3, pp. 544–549. © Pleiades Publishing, Inc., 2023. Russian Text © The Author(s), 2023, published in Molekulyarnaya Biologiya, 2023, Vol. 57, No. 3, pp. 539–545. |
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| doi: |
10.1134/S0026893323030068 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 520 | |a 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 $ 10^{2} $–$ 10^{3} $ 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 $ H37_{rv} $, 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. | ||
| 700 | 1 | |a Surzhikov, S. A. |4 aut | |
| 700 | 1 | |a Blagodatskikh, S. A. |4 aut | |
| 700 | 1 | |a Shershov, V. E. |4 aut | |
| 700 | 1 | |a Chudinov, A. V. |4 aut | |
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