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

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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.

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The authors declare no conflict of interest.

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Authors and Affiliations

  1. 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

    Manel Ben M’hadheb & Myriam Harrabi

  2. Institut Supérieur de Biotechnologie de Monastir, Université de Monastir, Avenue Tahar Hadded, BP 74, 5000, Monastir, Tunisia

    Manel Ben M’hadheb & Jawhar Gharbi

  3. Laboratoire des Maladies Transmissibles, Faculté de Pharmacie de Monastir, Avenue Avicenne, 5000, Monastir, Tunisia

    Amira Souii, Nadia Jrad-Battikh & Jawhar Gharbi

  4. Institut Supérieur des Sciences Biologiques Appliquées de Tunis, Université de Tunis El Manar, 9 Rue Zouhair Essafi, 1006, Tunis, Tunisia

    Amira Souii

Authors
  1. Manel Ben M’hadheb
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  2. Myriam Harrabi
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  3. Amira Souii
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  4. Nadia Jrad-Battikh
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  5. Jawhar Gharbi
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Correspondence to Jawhar Gharbi.

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Ben M’hadheb, M., Harrabi, M., Souii, A. et al. Multiplex RT-PCR and Indirect Immunofluorescence Assays for Detection and Subtyping of Human Influenza Virus in Tunisia. Curr Microbiol 70, 324–329 (2015). https://doi.org/10.1007/s00284-014-0719-0

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  • DOI: https://doi.org/10.1007/s00284-014-0719-0

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