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A Real Time PCR strategy for the detection and quantification of Candida albicans in human blood

2020; UNIVERSIDADE DE SÃO PAULO; Volume: 62; Linguagem: Inglês

10.1590/s1678-9946202062009

1678-9946

Felipe Delatorre Busser, Vivian Caso Coelho, Claudia de Abreu Fonseca, Gilda Maria Bárbaro Del Negro, Maria Aparecida Shikanai‐Yasuda, Marta Heloísa Lopes, Marcello Mihailenko Chaves Magri, Vera Lúcia Teixeira de Freitas,

Antimicrobial Resistance in Staphylococcus

Candidemia is a significant cause of bloodstream infections (BSI) in nosocomial settings. The identification of species can potentially improve the quality of care and decrease human mortality. Quantitative PCR (qPCR) was evaluated for Candida albicans detection using culture suspensions containing C. albicans , spiked human blood, the cloned qPCR target fragment (ITS2 region) and the results of these assays were compared. The assays showed a good detection limit: C. albicans DNA extracted from yeast (sensitivity 0.2 CFU/µL), spiked human blood (sensitivity 10 CFU/mL), and cloned fragment of ITS2 region (sensitivity 20 target copies/μL). The efficiency of ITS2 fragment-qPCR ranged from 89.67 to 97.07, and the linearity (R2) of the standard curve ranged from 0.992 to 0.999. The results showed that this ITS2-qPCR has a great potential as a molecular prototype model for the development of a test to be applied in clinical practice, greatly reducing the time of candidemia diagnosis, which is extremely important in this clinical setting.