Abstract

REYES O, Jesús; COMACH P, Guillermo; FRANCO, Leticia  and  CAMACHO G, Daría. STANDARIZATION OF MOLECULAR TECHNIQUES FOR DIAGNOSIS OF FLAVIVIRUS AND ALPHAVIRUS. Comunidad y Salud [online]. 2016, vol.14, n.1, pp.27-32. ISSN 1690-3293.

Flavivirus and alphavirus affect the health of humans. In Venezuela and particularly in Aragua, the communities have been affected by some members of these genus, such as dengue (DENV), and Chikungunya (CHIKV). DENV circulating in Aragua since 1989 generating outbreaks of clinical importance, while CHIKV made its appearance in 2014. In Aragua state, the diagnosis had been directed toward the detection of DENV, however the situation with CHIKV generates the need to expand the diagnosis spectrum to other viral agents. In the LARDIDEV we adapted two methodology of RT-PCR´s previously de scribed to detect members of these genus using flavivirus (DENV y Zika) and alphavirus (CHIKV) control strains. Both techniques were modified in the concentration of some reactants (MgCl2, dNTP´s, and primers) in the second reaction of PCR. The other conditions are kept equal to those originally described. Both techniques allowed amplified up to 1 fg of RNA of viral strains (Zika and CHIKV virus), but it could to amplify smaller amounts. In all cases, sharp bands according to the expected sizes using previously identified strains of DENV, CHIKV and Zika virus were obtained. The implementation of these methodologies will strengthen the timely diagnosis of members of this genus within the system of epidemiological surveillance of viral diseases.

Keywords : Flavivirus; Alphavirus; Zika; Dengue; Chikungunya; Molecular diagnosis.

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