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


ABSTRACT: 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/?L). 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.

SUBMITTER: Busser FD 

PROVIDER: S-EPMC7014565 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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

Busser Felipe Delatorre FD   Coelho Vivian Caso VC   Fonseca Claudia de Abreu CA   Del Negro Gilda Maria Barbaro GMB   Shikanai-Yasuda Maria Aparecida MA   Lopes Marta Heloisa MH   Magri Marcello Mihailenko Chaves MMC   Freitas Vera Lucia Teixeira de VLT  

Revista do Instituto de Medicina Tropical de Sao Paulo 20200207


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  ...[more]

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