Abstract

PADILLA R, Carlos; MONTOYA P, Ysabel  and  CARRILLO P, Carlos. Estandarización de una prueba de PCR  para la detección de Brucella sp. . Rev. perú. med. exp. salud publica [online]. 2003, vol.20, n.2, pp.102-104. ISSN 1726-4634.

Objetive: To standardize a PCR assay for detecting Brucella spp. Materials and methods: Reported primers targeted at the 16S rRNA gene of Brucella were used. Two DNA isolation methods were assessed: phenol-chloroform-isoamyl alcohol method and a comercial kit based on DNA affinity column. Eight Peruvian isolates of Brucella spp. were used to assess sensitivity and 7 different bacterial isolates, including E. coli, Shigella, Proteus mirabilis, Salmonella paratyphi, Salmonella typhi, Citrobacter freundii and Vibrio cholerae were used to assess specificity. Results: Both DNA isolation methods were effective for DNA isolation. The analytical sensitivity of the test was high as we were able to detect at least 80 femtograms of purified DNA. All isolates of Brucella spp. were detected by the assay. Furthermore, the test was negative with 7 bacteria species evaluated in this study. Conclusion: The conditions of a PCR assay to detect Peruvian isolates of Brucella spp, which has been shown to be very sensitive and specific in the laboratory have been tandardized.

Keywords : Polymerase Chain Reaction; Brucella; DNA.

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