Affiliation:
1. Institute of Biomedical Engineering1 and
2. Department of Pathology, National Cheng Kung University Hospital,2 and
3. Department of Clinical Pathology, Linko Medical Center, Chang Gung Memorial Hospital,3 Tainan 701, Taiwan, Republic of China
4. Department of Medical Technology, College of Medicine,4 National Cheng Kung University,
Abstract
ABSTRACT
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.
Publisher
American Society for Microbiology
Cited by
92 articles.
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