Publication: Assessment of a PCR technique for the detection and identification of cryptococcus neoformans
Issued Date
1996-12-01
Resource Type
ISSN
14602709
13693786
13693786
Other identifier(s)
2-s2.0-79961159291
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Mahidol University
Rights Holder(s)
SCOPUS
Bibliographic Citation
Medical Mycology. Vol.34, No.4 (1996), 251-258
Suggested Citation
C. Prariyachatigul, A. Chaiprasert, V. Meevootisom, S. Pattanakitsakul Assessment of a PCR technique for the detection and identification of cryptococcus neoformans. Medical Mycology. Vol.34, No.4 (1996), 251-258. doi:10.1080/02681219680000431 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/17686
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Title
Assessment of a PCR technique for the detection and identification of cryptococcus neoformans
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Abstract
The 18S ribosomal RNA gene of cryptococcus neoformans was amplified by polymerase chain reaction (PcR). The primers cPL1 and cPR4 were tested for their ability to amplify DNA from 30 strains of c. neoformans and 27 specimens of cerebrospinal fluid (cSF) from patients with cryptococcal meningitis. A 343 bp product was obtained and its specificity confirmed by Southern hybridization with an internal sequence (INSR4) probe. The sensitivity was 100 fg by Southern analysis and 1 pg using the PcR. Neither human nor a variety of other fungal and bacterial strains (n = 78) gave an amplified product. This PcR method can detect as few as 5 cells ml-1 of c. neoformans in spiked-cSF following a simple processing procedure. The developed system of PcR was more sensitive than the culture method and revealed a very high specificity. The PcR was easy to perform and needed only 4 h for all processes from receiving the cSF to detection of a specific DNA band after agarose gel electrophoresis. This would provide another rapid laboratory method for the diagnosis of cryptococcal meningitis. © 1996 Informa UK Ltd All rights reserved.