title: PCR-restriction fragment length polymorphism analysis of the phospholipase B (PLB1) gene for subtyping of Cryptococcus neoformans isolates.
abstract: 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.
authors.affiliation: Molecular Mycology Laboratory, Westmead Hospital, New South Wales, Australia.
authors.forename: G Nicolas
authors.initials: GN
authors.lastname: Latouche
authors.forename: Matthew
authors.initials: M
authors.lastname: Huynh
authors.forename: Tania C
authors.initials: TC
authors.lastname: Sorrell
authors.forename: Wieland
authors.initials: W
authors.lastname: Meyer
mesh_terms: D003455:Cryptococcus neoformans / Q000145:classification* / Q000201:enzymology / Q000235:genetics; D015247:Deoxyribonuclease HindIII; D015252:Deoxyribonucleases, Type II Site-Specific; D005656:Fungal Proteins / Q000235:genetics; D006801:Humans; D008245:Lysophospholipase / Q000235:genetics*; D016533:Mycological Typing Techniques; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012703:Serotyping
chemical_list: D005656:Fungal Proteins; D008245:Lysophospholipase; D015247:Deoxyribonuclease HindIII; C028678:CYCGRG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific