Multiplex PCR for Identification of Bacterial Pathogens of Infectious Pneumonia
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Multiplex PCR for Identification of Bacterial Pathogens of Infectious Pneumonia S. A. Lapaa, 1, E. S. Klochikhinaa, R. A. Miftakhova, A. M. Zolotova, A. S. Zasedateleva, and A. V. Chudinova aEngelhardt
Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia Received April 4, 2020; revised April 16, 2020; accepted April 20, 2020
Abstract—A multiplex PCR system has been developed and optimized for rapid detection of the five main pathogens of bacterial pneumonia. The system can be expanded to analyze viral pathogens of pneumonia (DNA- and RNA-containing viruses), as well as those of a fungal nature. Keywords: infectious pneumonia, multiplex PCR, diagnostics DOI: 10.1134/S1068162020050131
INTRODUCTION Infectious pneumonia is an acute inflammatory process of the lung tissue caused by a wide range of pathogens of bacterial, viral and fungal nature. Outbreaks of infectious pneumonia caused by SARS-CoV coronaviruses (2002–2003 yr), MERS-CoV (2012– 2013 yr), SARS-CoV-2 (2019–? yr), Influenza A and B viruses, as well as bacterial pneumonia, can become a real social threat. Bacterial pneumonia is characterized by a mortality rate of about 15%, while mortality in cases of the severe course of community-acquired pneumonia (SAP) can reach 21–58% [1], which is comparable to those of SARS (so-called atypical pneumonia), MERS [2] and exceeds this indicator for CoViD-19 and influenza. Specialized medical institutions that receive patients with a clinical diagnosis of pneumonia are faced with the problem of establishing the etiology of the disease and rapid identification of the pathogen, since often viral and bacterial pneumonia are characterized by a similar clinical picture. The situation sharply worsened with the onset of the CoViD-19 pandemic, which requires timely differential diagnosis of a secondary bacterial infection, which often arises as one of the complications of the course of the disease. The correct selection of drugs and the patient’s treatment strategy depend on the timely diagnosis [3]. The current situation signals an urgent need to create express methods of differential diagnostics. Abbreviations: SARS, severe acute respiratory syndrome; SAP, severe community-acquired pneumonia; CTAB, cetyl-trimethyl ammonium bromide. 1 Corresponding author: phone: +7 (495) 135-98-00; fax: +7 (495) 135-14-05; e-mail: [email protected].
Multiplex PCR was developed for the species determination of the five main pathogens of bacterial pneumonia in humans. The system is focused on the use in clinical diagnostics to determine the etiology of the disease due to the ability to differentiate bacterial from viral and fungal pneumonia. RESULTS AND DISCUSSION One of the most common pathogens of bacterial pneumonia are representatives of several genera: Staphylococcus aureus, Pseudomonas aeruginosa, Haemophilus influenza, Legionella pneumophila, Klebsiella pneumonia and some others [4]. For these five bacterial species, genetic targets were selected and primers were designed to perform multiplex
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