Revista Medica Herediana
versión On-line ISSN 1729-214X
HENRIQUEZ, César et al. Identificación de Bartonella bacilliformis por métodos moleculares. Rev Med Hered [online]. 2002, vol.13, n.2, pp. 58-63. ISSN 1729-214X.
Background: The PCR methodology amplifies a sequence of DNA with the Polimerase enzyme; the sensibility and specificity is very high. Objective: To develop a PCR methodology designed to work on extracts from whole blood samples rather than from isolated, cultured organisms. Methods: The whole blood of six patients with clinical and microbiologic diagnosis of acute bartonelosis by B. bacilliformis was used. The DNA was extracted with DNAzol® BD (lysis solution with guanidine detergent), and the extract subjected to PCR using primers 16S and 23S for the Intergenic Transcribed Spacer (ITS). The amplified products were subjected to electrophoresis on 1% agarose gels. Results: The concentration of the extracted product with DNAzol® BD was around 6 ng. The amplified single sized product of 1000 base pairs was identical to that from authentic cultures of B. bacilliformis and clearly different from that of other species. The dilutions detected by PCR were 1/5 and 1/10. Conclusion: The amplification of the DNA of B. bacilliformis extracted with DNAzol® BD from whole blood of patients with acute bartonelosis using the primers 16S and 23S is possible using this method for a fast etiologic diagnosis.
Palabras llave : PCR; Bartonella bacilliformis; DNAzol® BD; 16S 23S.