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Inclusion of Quality Controls on Leishmaniases Molecular Tests to Increase Diagnostic Accuracy in Research and Reference Laboratories

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Abstract

Early detection of leishmaniases and prompt institution of treatment are paramount for individuals and communities affected by these diseases. To overcome the remaining limitations inherent to molecular methods currently used and to ensure the accuracy of results in leishmaniases diagnosis, two triplex polymerase chain reaction (PCR) assays with quality controls for the reactions were developed. Validity indicators were assessed in 186 dog blood samples from endemic areas in Brazil. The level of agreement between the new tools and their singleplex protocols was assessed by kappa analysis. The triplex PCR for visceral leishmaniasis showed sensitivity (S) = 78.68 %, specificity (E) = 85.29 %, and efficiency (e) = 81.05 %. The cutaneous leishmaniasis protocol showed S = 97.29 %, E = 79.16 %, and e = 90.16 %. Both protocols showed good agreement with gold standards. These new tools enable, in a single reaction, the diagnosis of the diseases and the evaluation of the sample quality and DNA extraction process, thus reducing the cost of reagents and avoiding the eventual need for collecting a second sample.

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Acknowledgments

We appreciate Claudia Cafarchia´s help by sending the M. pachydermatis material from the Department of Veterinary Medicine, University of Bari, Italy; and Luciana Aguiar Figueredo, Filipe Dantas-Torres, and Domenico Otranto by the scientific opinion. Thanks to the Program of Technological Development in Health Supplies (PDTIS/FIOCRUZ) for allowing us to use its facilities, and to the State of Pernambuco Research Foundation (FACEPE), the National Council for Scientific and Technological Development (CNPq)/PAPES VI and Coordination for the Improvement of Higher Education Personnel (CAPES) for financial support.

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The authors declare no conflicts of interest in this publication.

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Authors and Affiliations

  1. Department of Immunology, Aggeu Magalhães Research Center, Oswaldo Cruz Foundation, Av. Prof. Moraes Rego, s/n, Recife, Pernambuco, 50670-420, Brazil

    Suênia da C. Gonçalves-de-Albuquerque, Rômulo Pessoa-e-Silva, Lays A. M. Trajano-Silva, Rayana C. S. de Morais, Sinval P. Brandão-Filho & Milena de Paiva-Cavalcanti

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  1. Suênia da C. Gonçalves-de-Albuquerque
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  2. Rômulo Pessoa-e-Silva
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  6. Milena de Paiva-Cavalcanti
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Correspondence to Milena de Paiva-Cavalcanti.

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da C. Gonçalves-de-Albuquerque, S., Pessoa-e-Silva, R., Trajano-Silva, L.A.M. et al. Inclusion of Quality Controls on Leishmaniases Molecular Tests to Increase Diagnostic Accuracy in Research and Reference Laboratories. Mol Biotechnol 57, 318–324 (2015). https://doi.org/10.1007/s12033-014-9825-2

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  • DOI: https://doi.org/10.1007/s12033-014-9825-2

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