USE OF MULTIPLEX PCR METHOD WITH REAL-TIME DETECTION FOR DIFFERENTIAL DIAGNOSIS OF RESPIRATORY VIRAL INFECTIONS


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Abstract

Multiplex real-time polymerase chain reaction test-system with fluorescent detection (RT-PCR) for simultaneous identification of main agents of acute respiratory viral infections: influenza A (IAV) and B viruses (IBV), parainfluenza viruses types 1, 2, 3, 4 (PIV 1 — 4), adenoviruses (ADV), respiratory syncitial virus (RSV), rhinoviruses (RV) and enteroviruses (EV), in presence internal positive control (IPC) represented by vaccine strain of rubella virus RA 27/3. Using multiplex RT-PCR method, respiratory viruses were detected in 116 out of 226 clinical samples (nasal swabs) obtained from patients with symptoms of acute respiratory infection: in 68 (58.6%) samples — IBV; in 21 (18.1%) — IAV; in 12 (10.3%) — RV; in 6 (5.2%) — PIV 2; in 4 — (3.4%) ADV; in 3 (2.6%) — RSV; in 2 (1.7%) — EV; in 2 (1.7%) — PIV 4; in 1 (0.9%) — PIV 3; in 1 (0.9%) — PIV 1. Mixed infection was observed in 4 (3.4%) patients. PCR assay allowed to reveal various respiratory viruses in 51.3% of samples. At the same time samples were tested for the presence of 12 respiratory viruses — IAV, IBV, PIV 1 — 4, RSV, RV, metapneumoviruses, and coronaviruses NL63, 229E and OC43 — in the presence of IPC represented by equine arteritis virus using analogous PCR test-system provided by medical center of Leiden university. Results of tests for detection of IAV, IBV, RSV, PIV 1 — 4, and RV, analyzed by both systems, agreed in 94%. Multiplex format of RT-PCR performing significantly reduces time and cost of the test, which make it suitable and effective instrument of epidemiological studies.

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ПРИМЕНЕНИЕ МЕТОДА МУЛЬТИПЛЕКСНОЙ ПЦР С ДЕТЕКЦИЕЙ В РЕЖИМЕ РЕАЛЬНОГО ВРЕМЕНИ ДЛЯ ДИФФЕРЕНЦИАЛЬНОЙ ДИАГНОСТИКИ РЕСПИРАТОРНЫХ ВИРУСНЫХ ИНФЕКЦИЙ
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About the authors

A. A Nikonova

Mechnikov Research Institute of Vaccines and Sera, Moscow; Center of Hygiene and Epidemiology in Moscow region, Mytishchi, Russia; Medical Center of Leiden University, Netherlands

E. S Uspenskaya

Mechnikov Research Institute of Vaccines and Sera, Moscow; Center of Hygiene and Epidemiology in Moscow region, Mytishchi, Russia; Medical Center of Leiden University, Netherlands

S. A Lobodanov

Mechnikov Research Institute of Vaccines and Sera, Moscow; Center of Hygiene and Epidemiology in Moscow region, Mytishchi, Russia; Medical Center of Leiden University, Netherlands

A. S Oksanich

Mechnikov Research Institute of Vaccines and Sera, Moscow; Center of Hygiene and Epidemiology in Moscow region, Mytishchi, Russia; Medical Center of Leiden University, Netherlands

A. E Gorbalenya

Mechnikov Research Institute of Vaccines and Sera, Moscow; Center of Hygiene and Epidemiology in Moscow region, Mytishchi, Russia; Medical Center of Leiden University, Netherlands

C. J.Claa Eric

Mechnikov Research Institute of Vaccines and Sera, Moscow; Center of Hygiene and Epidemiology in Moscow region, Mytishchi, Russia; Medical Center of Leiden University, Netherlands

E. P Foshina

Mechnikov Research Institute of Vaccines and Sera, Moscow; Center of Hygiene and Epidemiology in Moscow region, Mytishchi, Russia; Medical Center of Leiden University, Netherlands

A. N Kaira

Mechnikov Research Institute of Vaccines and Sera, Moscow; Center of Hygiene and Epidemiology in Moscow region, Mytishchi, Russia; Medical Center of Leiden University, Netherlands

V. V Zverev

Mechnikov Research Institute of Vaccines and Sera, Moscow; Center of Hygiene and Epidemiology in Moscow region, Mytishchi, Russia; Medical Center of Leiden University, Netherlands

E. B Fayzuloev

Mechnikov Research Institute of Vaccines and Sera, Moscow; Center of Hygiene and Epidemiology in Moscow region, Mytishchi, Russia; Medical Center of Leiden University, Netherlands

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Copyright (c) 2009 Nikonova A.A., Uspenskaya E.S., Lobodanov S.A., Oksanich A.S., Gorbalenya A.E., Eric C.J., Foshina E.P., Kaira A.N., Zverev V.V., Fayzuloev E.B.

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