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Journal of Clinical Microbiology, October 2001, p. 3466-3471, Vol. 39, No. 10
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.10.3466-3471.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Rapid Identification of Yeasts in Positive Blood Cultures by a Multiplex PCR Method

Hsein Chang Chang,1 Shiang Ning Leaw,1 Ay Huey Huang,2 Tsu Lan Wu,3 and Tsung Chain Chang4,*

Institute of Biomedical Engineering1 and Department of Medical Technology, College of Medicine,4 National Cheng Kung University, Department of Pathology, National Cheng Kung University Hospital,2 and Department of Clinical Pathology, Linko Medical Center, Chang Gung Memorial Hospital,3 Tainan 701, Taiwan, Republic of China

Received 6 December 2000/Returned for modification 30 April 2001/Accepted 20 July 2001

Yeasts are emerging as important etiological agents of nosocomial bloodstream infections. A multiplex PCR method was developed to rapidly identify clinically important yeasts that cause fungemia. The method amplified the internal transcribed spacer 1 (ITS1) region between the 18S and 5.8S rRNA genes and a specific DNA fragment within the ITS2 region of Candida albicans. With this method, C. albicans produced two amplicons, whereas other species produced only one. Through sequence analysis, the precise lengths of the PCR products were found to be as follows: C. glabrata (482 or 483 bp), C. guilliermondii (248 bp), C. parapsilosis (229 bp), C. albicans (218 or 219 and 110 bp), C. tropicalis (218 bp), Cryptococcus neoformans (201 bp), and C. krusei (182 bp). The PCR products could be effectively separated by disk polyacrylamide gel electrophoresis. The method was used to test 249 positive blood cultures (255 isolates), from which the following species (strain number) were isolated: C. albicans (128), C. tropicalis (51), C. glabrata (28), C. parapsilosis (23), C. neoformans (9), C. krusei (5), C. guilliermondii (3), and other, minor species (8). The test sensitivity of the method was 96.9% (247 of 255 isolates). The eight minor species were either misidentified (one strain) or not identified (seven strains). From the time at which a positive bottle was found, the multiplex PCR could be completed within 8 h; the present method is simpler than any previously reported molecular method for the identification of blood yeasts.

* Corresponding author. Mailing address: Department of Medical Technology, College of Medicine, National Cheng Kung University, 1 University Rd., Tainan 701, Taiwan, Republic of China. Phone: 886-6-235-3535, ext. 5790. Fax: 886-6-236-3956. E-mail: tsungcha{at}mail.ncku.edu.tw.

Journal of Clinical Microbiology, October 2001, p. 3466-3471, Vol. 39, No. 10
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.10.3466-3471.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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