Open AccessPCR-Restriction Fragment Length Polymorphism Analysis of the Phospholipase B ( PLB1 ) Gene for Subtyping of Cryptococcus neoformans Isolates
Author(s) -
GN Latouche,
Matthew Huynh,
Tania C. Sorrell,
Wieland Meyer
Publication year - 2003
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.69.4.2080-2086.2003
Subject(s) - serotype , cryptococcus neoformans , subtyping , restriction fragment length polymorphism , biology , restriction enzyme , genetics , polymorphism (computer science) , gene , microbiology and biotechnology , genotype , computer science , programming language
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.