![]() ![]() The major function of NA is to remove sialic acid residues that are present on newly synthesised virions and the membrane of infected cells. The hemagglutinin (HA) and neuraminidase (NA) glycoproteins are the main targets of the humoral immune response raised after natural infection or vaccination, whereas NA is a target of the main antiviral drugs licensed for the treatment of human influenza infection. Influenza A and B viruses are major causes of respiratory tract infections in humans. Monitoring antiviral drug resistance in viruses and developing molecular methods for early diagnosis are considered to be of a priority level similar to that of the development of methods to detect bacterial strains with antimicrobial resistance. We also discuss the advantage of using an RT-ddPCR instead of qPCR method in the context of viral diagnostics and surveillance. Furthermore, reverse genetics viruses carrying this mutation were created to test the performance of the RT-ddPCR assay and compare it to the standard phenotypic NA assay. In this study, based on an existing reverse transcriptase real-time PCR (RT-qPCR) assay, we developed a reverse transcriptase droplet digital PCR assay (RT-ddPCR) to detect and quantify the frequency of the E119V-NA mutation. Once a mutation such as E119V-NA is known, highly sensitive PCR-based genotypic assays can be used to identify the prevalence of such mutant influenza viruses in clinical samples. The neuraminidase inhibition assay allows the phenotypical identification of resistant strains however, this test often has limited sensitivity with high variability depending on the virus strain, drugs and assays. Early detection of resistant influenza viruses is important for patient management and for the rapid containment of antiviral resistance. Naturally occurring oseltamivir-resistant seasonal H3N2 influenza virus strains often carry a glutamate-to-valine substitution at position 119 in the neuraminidase (E119V-NA). ![]() The monitoring of antiviral-resistant influenza virus strains is important for public health given the availability and use of neuraminidase inhibitors and other antivirals to treat infected patients. ![]()
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