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J Clin Lab Anal. 2020 Feb 11:e23226. doi: 10.1002/jcla.23226. [Epub ahead of print]

Rapid identification of the Candida glabrata species complex by high-resolution melting curve analysis.

Author information

1
Department of Clinical Laboratory, Jinhua Municipal Central Hospital, Zhejiang, China.
2
State Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
3
School of Public Health, Wuhan University of Science and Technology, Hubei, China.
4
Department of Dermatology, Beijing Hospital, National Center of Gerontology, Beijing, China.

Abstract

BACKGROUND:

Candida glabrata is a common pathogen that causes invasive candidiasis. Among non-albicans Candida infections, C glabrata infections are associated with the highest fatality rates. Candida glabrata sensu stricto, Candida nivariensis, and Candida bracarensis have been identified and together form the C glabrata species complex. It is difficult to detect the two rare species by traditional laboratory methods. This study established a method for the rapid identification of members of the C glabrata species complex based on high-resolution melting curve (HRM) analysis and evaluated its practical application.

METHODS:

The internal transcribed spacer (ITS) region was used as target gene region to design specific primers. HRM analysis was performed with three subspecies of the C glabrata species complex and negative controls to test its specificity and sensitivity. To evaluate its practical application, the HRM technique was tested with clinical isolates, and the results were compared with the DNA sequencing results.

RESULTS:

Differences were detected among the melting profiles of the members of the C glabrata species complex. The negative controls were not amplified, indicating the high specificity of the method. The minimum detection limits of C glabrata sensu stricto, C nivariensis, and C bracarensis were approximately 1 × 101  copies/µL or less. The results of the HRM analysis of the clinical isolates were consistent with the DNA sequencing results.

CONCLUSIONS:

The HRM method is sensitive and can be used to rapidly identify the members of the C glabrata species complex. The method can allow early and targeted treatment of patients with invasive candidiasis.

KEYWORDS:

Candida glabrata ; high-resolution melting curve; identification; internal transcribed spacer; molecular typing

PMID:
32048348
DOI:
10.1002/jcla.23226

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