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AC-ELISA and RT-PCR assays for the diagnosis of triatoma virus (TrV) in triatomines (Hemiptera: Reduviidae) species

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Abstract

Triatoma virus (TrV) is the only entomopathogenic virus found in triatomines. TrV replicates in cells of the midgut epithelium of triatomines, causing a high mortality rate and delayed development of the infected insect. In this work, we report an antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) and a reverse transcription-polymerase chain reaction (RT-PCR) assay for detection of TrV infection. For antiserum production, rabbits and hens where inoculated with purified TrV. Antiserum reactivity was checked by immunodiffusion, and its specificity was confirmed by western blot and AC-ELISA. Totally 90 fecal samples from T. infestans were analysed. AC-ELISA and RT-PCR results correlated well with transmission electron microscopy (EM) observations, which are considered the gold standard, with Kappa values of 0.73 for AC-ELISA and 0.93 for RT-PCR when compared with EM. Applications and complementary uses of the two techniques reported in this work are discussed.

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Acknowledgments

GAM thanks the staff of Cátedra de Virología, FCV-UNLP, and the Programa de Movilidad de Profesores de la UPV/EHU. During this work, DMAG was a visiting professor of the UPV/EHU. JJG is a Researcher of the CICPBA; MGE is a Researcher of the CONICET. This study was partially supported by CONICET and Agencia Nacional de Promoción Científica y Tecnológicas, Argentina (PICT No 32618/05). We are very gratefully to Dr Eduardo Mórtola FCV-UNLP and Dr Félix Goñi (CSIC-UPV/EHU) for valuable comments on a draft of this manuscript. We are indebted to Lic G. Rozas-Dennis (UNS, Bahía Blanca, Argentina) for providing TrV-free insects and triatomine feces for AC-ELISA and RT-PCR analyses.

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Authors and Affiliations

  1. Centro Regional de Investigaciones Científicas y Transferencias Tecnológicas La Rioja (CRILAR, CONICET) Entre Ríos and Mendoza, 5310, Anillaco, La Rioja, Argentina

    Gerardo A. Marti

  2. Centro de Estudios Parasitológicos y de Vectores (CEPAVE-CCT-La Plata-CONICET-UNLP) 2 #584, 1900, La Plata, Argentina

    Gerardo A. Marti & Juan J. García

  3. Cátedra de Virología de la Facultad de Ciencias Veterinarias (FCV-UNLP and CONICET), 60 and 118, CC 296, 1900, La Plata, Argentina

    Ester T. González & María G. Echeverría

  4. Unidad de Biofísica (CSIC-UPV/EHU), Barrio Sarriena S/N. 48940, Leioa, Vizcaya, Spain

    Ana R. Viguera & Diego M. A. Guérin

Authors
  1. Gerardo A. Marti
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  2. Ester T. González
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  3. Juan J. García
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  4. Ana R. Viguera
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  5. Diego M. A. Guérin
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  6. María G. Echeverría
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Corresponding authors

Correspondence to Gerardo A. Marti or Diego M. A. Guérin.

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Marti, G.A., González, E.T., García, J.J. et al. AC-ELISA and RT-PCR assays for the diagnosis of triatoma virus (TrV) in triatomines (Hemiptera: Reduviidae) species. Arch Virol 153, 1427–1432 (2008). https://doi.org/10.1007/s00705-008-0130-x

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  • DOI: https://doi.org/10.1007/s00705-008-0130-x

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