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Updated: May 19, 2026

Development of Multiplex Real-Time RT-qPCR Assays for the Detection of SARS-CoV-2, Influenza A/B, and MERS-CoV
Published on: November 10, 2023
[Methods of detection of selected respiratory viruses]
Ilona Stefańska1, Magdalena Romanowska, Lidia B Brydak
1Zakład Badania Wirusów Grypy. Krajowy Ośrodek ds. Grypy, Narodowy Instytut Zdrowia Publicznego - Państwowy Zakład Higieny w Warszawie. i.stefanska@gmail.com
Abstract:
Respiratory viruses contribute to significant morbidity and mortality in healthy and immunocompromised individuals and are considered as a significant economic burden in the healthcare system. The similar clinical symptoms in the course of different viral and bacterial respiratory infections make the proper diagnosis difficult. An accurate and prompt diagnostics is crucial for infection control and patient management decisions, especially regarding the use of antibacterial or antiviral therapy and hospitalization. Moreover, the identification of the causative agent eliminates inappropriate use of antibiotics and may reduce the cost of healthcare. A wide variety of diagnostic procedures is applied for the detection of viral agents responsible for respiratory tract infections. For many years, the viral antigen detection and standard isolation technique in cell culture was the main method used in routine diagnostics. However, in recent years the nucleic acid amplification techniques have become widely used and have significantly improved the sensitivity of viral detection in clinical specimens. Molecular diagnostic assays have contributed to revealing high rates of co-infection (multiplex reactions) and allow identification of agents that are difficult to culture. This paper discusses a number of technical aspects of the current most commonly used techniques, their general principles, main benefits and diagnostic value, but also some of their limitations.
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