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 co...

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Bibliographic Details
Published in:Revista do Instituto de Medicina Tropical de São Paulo
Main Authors: Felipe Delatorre Busser, Vivian Caso Coelho, Claudia de Abreu Fonseca, Gilda Maria Barbaro Del Negro, Maria Aparecida Shikanai-Yasuda, Marta Heloisa Lopes, Marcello Mihailenko Chaves Magri, Vera Lucia Teixeira de Freitas
Format: Article in Journal/Newspaper
Language:English
Published: Universidade de São Paulo (USP) 2020
Subjects:
Candida albicans
Candidemia
Detection
Invasive candidiasis
ITS2
Quantitative PCR
Arctic medicine. Tropical medicine
RC955-962
Infectious and parasitic diseases
RC109-216
Arctic
Online Access:https://doi.org/10.1590/s1678-9946202062009
https://doaj.org/article/bb7e8a6ba2ec4059a1ce8c391f8ff4ca
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Summary: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/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.

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