Utility of two PCR‐RFLP‐based techniques for identification of Candida parapsilosis complex blood isolates

Autor:	Emilia Cantón, Elena Eraso, Estibaliz Mateo, Cristina Marcos-Arias, Javier Pemán, Nerea Pena‐Fernández, Guillermo Quindós, Irene Jurado-Martín
Rok vydání:	2020
Předmět:	0301 basic medicine Species complex Candida parapsilosis 030106 microbiology Microbial Sensitivity Tests Dermatology Biology Polymerase Chain Reaction Microbiology 030207 dermatology & venereal diseases 03 medical and health sciences 0302 clinical medicine Candida metapsilosis CANDIDA ORTHOPSILOSIS Candida krusei Prevalence Humans Mycological Typing Techniques Candida albicans Phylogeny Candidemia Reproducibility of Results Sequence Analysis DNA General Medicine Ribosomal RNA biology.organism_classification Infectious Diseases RNA Ribosomal Restriction fragment length polymorphism Polymorphism Restriction Fragment Length
Zdroj:	Mycoses. 63:461-470
ISSN:	1439-0507 0933-7407
DOI:	10.1111/myc.13061
Popis:	Background Candida parapsilosis is the second or third most frequently isolated Candida species related to nosocomial infections, even overtaking Candida albicans in some hospitals. C. parapsilosis constitutes a complex of closely related species: Candida parapsilosis sensu stricto, Candida orthopsilosis and Candida metapsilosis. Accurate detection of these species is of importance, as the incidence of C. orthopsilosis has been reported to surpass that of Candida krusei. Objective To evaluate the diagnostic utility of two PCR-RFLP methods targeting the SADH and FKS1 genes and to determine the prevalence of cryptic species in 96 bloodstream isolates of C. parapsilosis from 93 patients. Methods Restriction patterns of the SADH and FKS1 genes were analysed, and sequencing of the D1/D2 regions of the ribosomal RNA was used to evaluate the reliability of both PCR-RFLP methods. Results In our study, 77 C. parapsilosis sensu stricto, 13 C. orthopsilosis and five C. metapsilosis were identified by sequencing. Both PCR-RFLP methods demonstrated strong agreement with D1/D2 sequencing in the identification of C. parapsilosis and C. orthopsilosis, while both methods were unable to identify the C. metapsilosis isolates. Moreover, unexpected restriction patterns were observed for two isolates on SADH PCR-RFLP and for four isolates on FKS1 PCR-RFLP. Mixed bloodstream infections of C. parapsilosis sensu stricto and C. orthopsilosis were detected for three patients, for which differential growth characteristics were observed. Conclusion The molecular method chosen for identification could have an impact on determination of the real prevalence of C. metapsilosis in candidaemia, and mixed fungaemias can remain undetected.
Databáze:	OpenAIRE
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