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Journal of Clinical Microbiology, October 2001, p. 3466-3471, Vol. 39, No. 10
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.
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
*
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.
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