Acta Medica Colombiana
Print version ISSN 0120-2448
OSPINA, William Pascual and CORTES, Jorge Alberto. Molecular diagnosis of candidemia by seminested PCR in critically ill patients. Acta Med Colomb [online]. 2011, vol.36, n.3, pp. 135-140. ISSN 0120-2448.
Objectives: to standardize a molecular test based on semi-nested PCR for diagnosis of candidemia. Methods: we developed a pilot study by means of optimizing the extraction protocol and use of ribosomal DNA, amplifying the complete ITS-2 region of Candida spp. And by semi-nested PCR reamplifying specific segments of nine clinically relevant Candida species. Clinical samples of plasma of critically ill patients with microbiologically proven candidemia from a third level hospital in Bogotá. Another sample of simulated candidemia was used with Candida strain form a urine sample. Results: we were able to standardize this technique and achieved from clinical samples of plasma a likely sensitivity of 90% and a theoretical specificity of 100%. The detection capability of this test is up to 1 cell per 200 μl of plasma. Conclusion: a molecular test for detection of candidemia among critically ill patients was standardized (Acta Med Colomb 2011; 36: 135-140).
Keywords : (DeCS); candidemia/microbiology; candida; candida/isolation & purification; candidemia; candidemia/diagnosis; candidemia/microbiology; molecular diagnostic technology; polymerase chain reaction; nested PCR.