title: Easy and inexpensive molecular detection of dengue, chikungunya and zika viruses in febrile patients.
abstract: Dengue (DENV), chikungunya (CHIKV) and zika (ZIKV) are arthropod-borne viruses (arboviruses) sharing a common vector, the mosquito Aedes aegypti. At initial stages, patients infected with these viruses have similar clinical manifestations, however, the outcomes and clinical management of these diseases are different, for this reason early and accurate identification of the causative virus is necessary. This paper reports the development of a rapid and specific nested-PCR for detection of DENV, CHIKV and ZIKV infection in the same sample. A set of six outer primers targeting the C-preM, E1, and E gene respectively was used in a multiplex one-step RT-PCR assay, followed by the second round of amplification with specific inner primers for each virus. The specificity of the present assay was validated with positive and negative serum samples for viruses and supernatants of infected cells. The assay was tested using clinical samples from febrile patients. In these samples, we detected mono and dual infections and a case of triple co-infection DENV-CHIKV-ZIKV. This assay might be a useful and an inexpensive tool for detection of these infections in regions where these arboviruses co-circulate.
authors.affiliation: Laboratorio de Virología, Universidad El Bosque, Bogotá, Colombia. Electronic address: calvoeliana@unbosque.edu.co.
authors.forename: Eliana P
authors.initials: EP
authors.lastname: Calvo
authors.affiliation: Laboratorio de Virología, Universidad El Bosque, Bogotá, Colombia; Grupo Patogénesis Infecciosa, Universidad Nacional de Colombia, Colombia.
authors.forename: Fernando
authors.initials: F
authors.lastname: Sánchez-Quete
authors.affiliation: Laboratorio de Virología, Universidad El Bosque, Bogotá, Colombia.
authors.forename: Sandra
authors.initials: S
authors.lastname: Durán
authors.affiliation: Laboratorio de Virología, Universidad El Bosque, Bogotá, Colombia.
authors.forename: Isabel
authors.initials: I
authors.lastname: Sandoval
authors.affiliation: Laboratorio de Virología, Universidad El Bosque, Bogotá, Colombia; Grupo Patogénesis Infecciosa, Universidad Nacional de Colombia, Colombia.
authors.forename: Jaime E
authors.initials: JE
authors.lastname: Castellanos
mesh_terms: D065632:Chikungunya Fever / Q000175:diagnosis / Q000821:virology; D002646:Chikungunya virus / Q000302:isolation & purification; D017931:DNA Primers; D003715:Dengue / Q000175:diagnosis / Q000821:virology; D003716:Dengue Virus / Q000302:isolation & purification; D005334:Fever; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D012327:RNA Virus Infections / Q000175:diagnosis* / Q000821:virology; D012328:RNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D000071244:Zika Virus / Q000302:isolation & purification; D000071243:Zika Virus Infection / Q000175:diagnosis / Q000821:virology
chemical_list: D017931:DNA Primers
keywords: Arbovirus; Chikungunya; Dengue; Detection; Nested PCR; Zika