title: Multiplex RT-PCR and indirect immunofluorescence assays for detection and subtyping of human influenza virus in Tunisia.
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.
authors.affiliation: Laboratoire de Génétique et de Valorisation des Bioresources (LR11-ES41), Institut Supérieur de Biotechnologie de Monastir, Université de Monastir, Avenue Taher Hadded, BP 74, 5000, Monastir, Tunisia.
authors.forename: Manel
authors.initials: M
authors.lastname: Ben M'hadheb
authors.forename: Myriam
authors.initials: M
authors.lastname: Harrabi
authors.forename: Amira
authors.initials: A
authors.lastname: Souii
authors.forename: Nadia
authors.initials: N
authors.lastname: Jrad-Battikh
authors.forename: Jawhar
authors.initials: J
authors.lastname: Gharbi
mesh_terms: D000293:Adolescent; D000328:Adult; D002648:Child; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods; D005838:Genotype; D006801:Humans; D007251:Influenza, Human / Q000175:diagnosis / Q000821:virology*; D008875:Middle Aged; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D009975:Orthomyxoviridae / Q000145:classification* / Q000235:genetics*; D014416:Tunisia; D055815:Young Adult