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Applied and Environmental Microbiology, April 2003, p. 2080-2086, Vol. 69, No. 4
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.4.2080-2086.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

PCR-Restriction Fragment Length Polymorphism Analysis of the Phospholipase B (PLB1) Gene for Subtyping of Cryptococcus neoformans Isolates

G. Nicolas Latouche,^1,2,3 Matthew Huynh,^1,2,3 Tania C. Sorrell,^2,3 and Wieland Meyer^1,2,3*

Molecular Mycology Laboratory,¹ Centre for Infectious Diseases and Microbiology, Westmead Hospital, Westmead,² Department of Medicine, University of Sydney, Sydney, New South Wales, Australia³

Received 16 July 2002/ Accepted 6 January 2003

Cryptococcus neoformans is a pathogenic yeast that is currently divided into three varieties, five serotypes, and eight molecular types. The following report describes the use of PCR-restriction fragment length polymorphism (RFLP) analysis of the phospholipase B gene (PLB1) as a simple tool to differentiate between C. neoformans subgroups. A PLB1 fragment, 1,970 bp, was amplified and digested with either AvaI or HindIII. Both sets of profiles grouped the isolates into their respective varieties, but only the AvaI profiles allowed for the identification of the eight molecular types via the corresponding RFLP profiles A1 to A8. Digestion of the same fragments with HindIII resulted in RFLP profiles H1 to H5, which distinguished only between serotype A, AD, D, and B/C. Neither enzyme distinguished serotype B from serotype C. The serotype AD profile was a composite of the serotype A and D profiles. Further investigation showed that the serotype AD isolates used in this study are heterozygous, with one allele of PLB1 originating from a serotype A parent and the other from a serotype D parent.

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* Corresponding author. Mailing address: Molecular Mycology Laboratory, Westmead Hospital, ICPMR, Level 3 Room 3114A, Darcy Rd., Westmead, New South Wales 2145, Australia. Phone: (61-2) 9845 6895. Fax: (61-2) 9891 5317. E-mail: w.meyer{at}usyd.edu.au.

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Applied and Environmental Microbiology, April 2003, p. 2080-2086, Vol. 69, No. 4
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.4.2080-2086.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

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