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Memorias del Instituto de Investigaciones en Ciencias de la Salud

versão On-line ISSN 1812-9528

Resumo

BERNAL-VERA, Cynthia Magdalena; CARDOZO-SEGOVIA, Fátima María  e  MENDOZA-TORRES, Laura Patricia. Standardization of a nested RT-PCR technique for alphavirus detection. Mem. Inst. Investig. Cienc. Salud [online]. 2017, vol.15, n.2, pp.30-36. ISSN 1812-9528.  https://doi.org/10.18004/mem.iics/1812-9528/2017.015(02)30-036.

The genus Alphavirus consists of RNA viruses of which several species are responsible for human and animal diseases, such as chikungunya, Mayaro and equine encephalitis viruses, and are therefore considered a regional public health problem. In Paraguay, human infections have been reported by chikungunya, but more studies are needed to increase knowledge on the circulation and ecoepidemiology of alphaviruses. Reverse RNA transcription followed by a nested polymerase chain reaction (RT-PCR) is very useful as a diagnostic tool and in epidemiological surveillance. The objective of this study was to define optimal reaction conditions and to determine the limit of detection for a nested RT-PCR for generic alphavirus detection. The detection limit obtained, of 0,47 PFU/mL, indicate high sensitivity, and the possibility of applying the technique to human and animals samples of serum, cerebrospinal fluid, organs and mosquito pools. This work will serve as a basis for other studies of detection and identification of alphavirus species circulating in our country, which would help strengthen the surveillance and prevention.

Palavras-chave : Alphavirus; nested RT-PCR; standardization..

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