title: Detection of dengue, west Nile virus, rickettsiosis and leptospirosis by a new real-time PCR strategy.
abstract: Dengue virus (DENV) infection causes sudden fever along with several nonspecific signs and symptoms and in severe cases, death. DENV is transmitted to people by Aedes aegypti and Ae. albopictus mosquitoes, whose populations increase during rainy season. West Nile Virus (WNV), Rickettsia spp. and Leptospira spp. are fever-causing pathogens that share many of the initial symptoms of DENV infection and also thrive in the rainy season. Outbreaks in some regions may be due to any of these pathogens that can co-circulate. Plus, they are clinically indistinguishable until severe symptoms appear, even though these diseases should be treated differently. An effective differential diagnosis would help clinicians and vector control departments to make right decisions for control and treatment of these diseases. Therefore, we developed four different SYBR green (®) -based reverse transcription quantitative PCR (RT-qPCR) assays for simultaneous detection of DENV, WNV, Rickettsia spp. and Leptospira spp. The assay has been optimized to yield results in less than 1 h; and in order to reduce contamination risk, all reagents were premixed and lyophilized on 96 well plates and thus only requires the addition of water and total nucleic acids from the sample. Sensitivities of the assays were less than 100 copies of nucleic acid targeted for these four pathogens. Assays did not show cross reactivity with any of the four pathogens nor to human nucleic acids. We are presenting a sensitive and selective kit that detects four relevant pathogens from tropical regions, that is quick, cost-effective and easy to use.
authors.affiliation: Dirección de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ), Avenida Normalistas No. 800, Colinas de la Normal, C.P. 44270 Guadalajara, Jalisco Mexico.
authors.forename: Daniel
authors.initials: D
authors.lastname: García-Ruíz
authors.affiliation: Dirección de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ), Avenida Normalistas No. 800, Colinas de la Normal, C.P. 44270 Guadalajara, Jalisco Mexico.
authors.forename: Marco A
authors.initials: MA
authors.lastname: Martínez-Guzmán
authors.affiliation: Dirección de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ), Avenida Normalistas No. 800, Colinas de la Normal, C.P. 44270 Guadalajara, Jalisco Mexico.
authors.forename: Albertina
authors.initials: A
authors.lastname: Cárdenas-Vargas
authors.affiliation: Dirección de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ), Avenida Normalistas No. 800, Colinas de la Normal, C.P. 44270 Guadalajara, Jalisco Mexico.
authors.forename: Erika
authors.initials: E
authors.lastname: Marino-Marmolejo
authors.affiliation: Dirección de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ), Avenida Normalistas No. 800, Colinas de la Normal, C.P. 44270 Guadalajara, Jalisco Mexico.
authors.forename: Abel
authors.initials: A
authors.lastname: Gutiérrez-Ortega
authors.affiliation: Departamento de Infectología, Hospital Civil Fray Antonio Alcalde, Calle Hospital No. 270, El Retiro, 44200 Guadalajara, Jalisco Mexico.
authors.forename: Esteban
authors.initials: E
authors.lastname: González-Díaz
authors.affiliation: Departamento de Infectología, Hospital Civil Fray Antonio Alcalde, Calle Hospital No. 270, El Retiro, 44200 Guadalajara, Jalisco Mexico.
authors.forename: Rayo
authors.initials: R
authors.lastname: Morfin-Otero
authors.affiliation: Departamento de Infectología, Hospital Civil Fray Antonio Alcalde, Calle Hospital No. 270, El Retiro, 44200 Guadalajara, Jalisco Mexico.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Rodríguez-Noriega
authors.affiliation: Departamento de Infectología, Hospital Civil Fray Antonio Alcalde, Calle Hospital No. 270, El Retiro, 44200 Guadalajara, Jalisco Mexico.
authors.forename: Hector
authors.initials: H
authors.lastname: Pérez-Gómez
authors.affiliation: Dirección de Biotecnología Médica y Farmacéutica, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco (CIATEJ), Avenida Normalistas No. 800, Colinas de la Normal, C.P. 44270 Guadalajara, Jalisco Mexico.
authors.forename: Darwin
authors.initials: D
authors.lastname: Elizondo-Quiroga
keywords: Dengue; Leptospira; Real time PCR; Rickettsia; West Nile Virus