Performance of a Real-Time PCR Assay for the Detection of Five Candida Species in Blood Samples from ICU Patients at Risk of Candidemia

Author:

Felix Gabriel N.1,de Freitas Vera L. T.2ORCID,da Silva Junior Afonso R.3ORCID,Magri Marcello M. C.34,Rossi Flavia3,Sejas Odeli N. E.5ORCID,Abdala Edson5ORCID,Malbouisson Luiz M. S.6,Guimarães Thais7ORCID,Benard Gil1ORCID,Del Negro Gilda M. B.1ORCID

Affiliation:

1. Laboratory of Medical Mycology (LIM 53), Instituto de Medicina Tropical, Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (HC-FMUSP), São Paulo 05403-000, Brazil

2. Laboratório de Investigação Médica em Imunologia (LIM 48), Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (HC-FMUSP), São Paulo 05403-000, Brazil

3. Central Laboratory Division (LIM 03), Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (HC-FMUSP), São Paulo 05403-010, Brazil

4. Departamento de Moléstias Infecciosas e Parasitárias, Faculdade de Medicina, Universidade de São Paulo, São Paulo 05403-010, Brazil

5. Cancer Institute of São Paulo State (ICESP), Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (HC-FMUSP), São Paulo 01246-000, Brazil

6. Discipline of Anesthesiology, Faculdade de Medicina, Universidade de São Paulo, São Paulo 05403-010, Brazil

7. Infectious Diseases Department, Hospital do Servidor Público Estadual de São Paulo (IAMSPE), São Paulo 04029-000, Brazil

Abstract

The gold standard for diagnosing invasive candidiasis still relies on blood cultures, which are inefficient and time-consuming to analyze. We developed an in-house qPCR assay to identify the 5 major Candida species in 78 peripheral blood (PB) samples from ICU patients at risk of candidemia. Blood cultures and (1,3)-β-D-glucan (BDG) testing were performed concurrently to evaluate the performance of the qPCR. The qPCR was positive for DNA samples from all 20 patients with proven candidemia (positive PB cultures), showing complete concordance with Candida species identification in blood cultures, except for detection of dual candidemia in 4 patients, which was missed by blood cultures. Additionally, the qPCR detected Candida species in six DNA samples from patients with positive central venous catheters blood (CB) but negative PB cultures. BDG values were similarly high in these six samples and the ones with proven candidemia, strongly suggesting the diagnosis of a true candidemia episode despite the negative PB cultures. Samples from patients neither infected nor colonized yielded negative results in both the qPCR and BDG testing. Our qPCR assay was at least as sensitive as blood cultures, but with a shorter turnaround time. Furthermore, negative results from the qPCR provided strong evidence for the absence of candidemia caused by the five major Candida species.

Funder

São Paulo Research Foundation

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil

Publisher

MDPI AG

Subject

Plant Science,Ecology, Evolution, Behavior and Systematics,Microbiology (medical)

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