Different species of protozoa parasites from Leishmania genus lead to different forms of infections, ranging from cutaneous lesions with spontaneous healing and disfiguring mucocutaneous affections, such as American Tegumentary Leishmaniasis (ATL), to lethal forms with visceralization, as Visceral Leishmaniasis (VL). Leishmaniasis is a complex of neglected diseases worldwide distributed and endemic in more than 98 countries including the Americas and presents an overall prevalence of 1.2 million cases, according to the World Health Organization (WHO). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Ĭompeting interests: The authors have declared that no competing interests exist.
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Elisa Cupolillo-Coordenação de Aperfeiçoamento de Pessoal de Nível Superior-Brasil (CAPES)-Finance Code 001 CNPq-Researcher Fellow (302622/2017-9) Faperj-CNE-E26-202.569/2019 Apoio às Instituições Sediadas no estado do Rio de Janeiro - E-26/010.101083/2018 C Britto and OC Moreira – researcher fellows of CNPq ((308411/2017-0) and FAPERJ (CNE, JCNE- E-26/203.031/2018). This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: All relevant data are within the manuscript and its Supporting Information files.įunding: This work was supported by grants from Fiocruz (Inova Fiocruz – Edital Produtos Inovadores/VPPCB), CNPq and FAPERJ. Received: MaAccepted: AugPublished: October 12, 2020Ĭopyright: © 2020 Filgueira et al. PLoS Negl Trop Dis 14(10):Įditor: Claudia Ida Brodskyn, Centro de Pesquisa Gonçalo Moniz-FIOCRUZ/BA, BRAZIL (2020) Comparison and clinical validation of qPCR assays targeting Leishmania 18S rDNA and HSP70 genes in patients with American Tegumentary Leishmaniasis. With this approach, our main goal is to conclude the first step of a further multicenter study to propose the standardization of detection and quantification of Leishmania.Ĭitation: Filgueira CPB, Moreira OC, Cantanhêde LM, de Farias HMT, Porrozzi R, Britto C, et al. Following the methodology proposed herein, the results indicate the use of both qPCR assays, 18S rDNA and HSP70, to achieve a very good net sensitivity (98.5%) and specificity (100%), performing simultaneous or sequential testing, respectively. Additionally, we validated reactions by assaying 88 samples from patients presenting different clinical forms of leishmaniasis (cutaneous, mucosal, recent and old lesions), representing the diversity found in Brazil’s Amazon Region. In this sense, the purpose of the present work is to compare the performance of qPCR using two commonly used targets (18S rDNA and HSP70) with an internal control (RNAse P) in multiplex reactions. Moreover, a proper validation by the assay by the use of clinical samples is still required. Although many studies have been already published addressing the use of this technique, an improvement on these methodologies, including an analytical validation, standardization and data association is demanded. Molecular approaches, specially based on Real Time PCR (qPCR) technique, has been widely used to detect Leishmania infection and to quantify parasite load, once it is a simple, rapid and sensitive methodology, capable to detect low parasite concentrations and less prone to variability. The sensitivity of the method varies, and factors such as collection procedures interfere. Currently, ATL diagnosis is mainly made by parasite detection by microscopy. As a neglected disease, much effort is still needed in treatment and diagnosis. Leishmaniasis is a worldwide neglected disease, encompassing asymptomatic infections and different clinical forms, such as American Tegumentary Leishmaniasis (ATL) which is part of the complex of diseases caused by protozoan parasites from Leishmania genus, transmitted by sand fly vectors.