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Multiplex RT-PCR and Indirect Immunofluorescence Assays for Detection and Subtyping of Human Influenza Virus in Tunisia

tetano

Editor, Senior Moderator
Curr Microbiol. 2014 Nov 1. [Epub ahead of print]
Multiplex RT-PCR and Indirect Immunofluorescence Assays for Detection and Subtyping of Human Influenza Virus in Tunisia.
Ben M'hadheb M1, Harrabi M, Souii A, Jrad-Battikh N, Gharbi J.
Author information
Abstract

Influenza viruses are negative stranded segmented RNA viruses belonging to Orthomyxoviridae family. They are classified into three types A, B, and C. Type A influenza viruses are classified into subtypes according to the antigenic characters of the surface glycoproteins: hemagglutinin (H) and neuraminidase (N). The aim of the present study is to develop a fast and reliable multiplex RT-PCR technique for detecting simultaneously the subtypes A/H1N1 and A/H3N2 of influenza virus. Our study included 398 patients (mean age 30.33 ? 19.92 years) with flu or flu-like syndromes, consulting physicians affiliated with collaborating teams. A multiplex RT-PCR detecting A/H1N1 and A/H3N2 influenza viruses and an examination by indirect immunofluorescence (IFI) were performed. In the optimized conditions, we diagnosed by IFI a viral infection in 90 patients (22.6 %): 85 cases of influenza type A, four cases of influenza type B, and only one case of coinfection with types A and B. An evaluation of the technique was performed on 19 clinical specimens positive in IFI, and we detected eight cases of A/H3N2, five cases of A/H1N1, one case of influenza virus type A which is not an H1N1 nor H3N2, and five negative cases. Multiplex RT-PCR is a sensitive technique allowing an effective and fast diagnosis of respiratory infections caused by influenza viruses in which the optimization often collides with problems of sensibility.

PMID:
25366277
[PubMed - as supplied by publisher]

http://www.ncbi.nlm.nih.gov/pubmed/25366277
 
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