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Updated: Dec 25, 2025
Author Spotlight: Advancements in Multiplex Detection of Respiratory Viruses
Published on: November 10, 2023
A multiplex real-time PCR assay for detection of oseltamivir-resistant strains of influenza virus
Dawid Nidzworski1,2, Joanna Dobkowska1,3, Marcin Hołysz2,4
11Department of Recombinant Vaccines, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Gdansk, Poland.
Abstract:
Influenza is a contagious disease of humans and animals caused by viruses belonging to the Orthomyxoviridae family. The influenza A virus genome consists of negative sense, single-stranded, segmented RNA. Influenza viruses are classified into subtypes based on two surface antigens known as hemagglutinin (H) and neuraminidase (N). The main problem with influenza A viruses infecting humans is drug resistance, which is caused by antigenic changes. A few antiviral drugs are available, but the most popular is the neuraminidase inhibitor - oseltamivir. The resistance against this drug has probably developed through antigenic drift by a point mutation in one amino acid at position 275 (H275Y). In order to prevent a possible influenza pandemic it is necessary to develop fast diagnostic tests. The aim of this project was to develop a new test for detection of influenza A virus and determination of oseltamivir resistance/sensitivity in humans. Detection and differentiation of oseltamivir resistance/sensitivity of influenza A virus was based on real-time PCR. This test contains two TaqMan probes, which work at different wavelengths. Application of techniques like multiplex real-time PCR has greatly enhanced the capability for surveillance and characterization of influenza viruses. After its potential validation, this test can be used for diagnosis before treatment.
Insights
A new real-time PCR test detects influenza A virus and determines oseltamivir resistance. This rapid diagnostic tool aids in timely treatment decisions for influenza patients, combating drug resistance.
Area of Science:
- Virology
- Molecular Biology
- Infectious Diseases
Background:
- Influenza A virus, an Orthomyxoviridae family member, poses a pandemic threat.
- Antigenic drift leads to drug resistance, notably against neuraminidase inhibitors like oseltamivir.
- The H275Y mutation is linked to oseltamivir resistance in influenza A viruses.
Purpose of the Study:
- To develop a novel diagnostic test for influenza A virus detection.
- To determine oseltamivir resistance or sensitivity in human influenza A virus infections.
- To enable rapid diagnosis for informed treatment strategies.
Main Methods:
- Real-time polymerase chain reaction (PCR) was employed for detection and differentiation.
- The assay utilizes two TaqMan probes operating at distinct wavelengths.
- Multiplex real-time PCR enhances surveillance and characterization capabilities.
Main Results:
- A new real-time PCR test was successfully developed.
- The test differentiates influenza A virus strains based on oseltamivir resistance/sensitivity.
- The assay leverages multiplexing for efficient viral characterization.
Conclusions:
- The developed real-time PCR test facilitates rapid influenza A virus diagnosis.
- It enables simultaneous determination of oseltamivir resistance/sensitivity.
- This diagnostic tool supports clinical decision-making and pandemic preparedness.
