A multiplex real-time PCR assay for detection of oseltamivir-resistant strains of influenza virus

作者：Nidzworski Dawid^*; Dobkowska Joanna; Hoysz Marcin; Gromadzka Beata; Szewczyk Bogusaw

来源：Central European Journal of Biology, 2014, 9(6): 628-633.

DOI：10.2478/s11535-014-0296-z

• Influenza
• Detection
• Oseltamivir
• H275Y
• Real-time PCR

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.

• 出版日期2014-6