Consensus multi-locus sequence typing scheme forCryptococcus neoformansandCryptococcus gattii

Abstract

This communication describes the consensus multi-locus typing scheme established by the Cryptococcal Working Group I (Genotyping of Cryptococcus neoformans and C. gattii) of the International Society for Human and Animal Mycology (ISHAM) using seven unlinked genetic loci for global strain genotyping. These genetic loci include the housekeeping genes CAP59, GPD1, LAC1, PLB1, SOD1, URA5 and the IGS1 region. Allele and sequence type information are accessible at http://www.mlst.net/.

Cryptococcus neoformans isolates from Yaoundé human immunodeficiency virus-infected patients exhibited intra-individual genetic diversity and variation in antifungal susceptibility profiles between isolates from the same patient

“…grubii AFLP1/VNI Â C. gattii AFLP4/VGI (serotype AB, genotype AFLP9) and C. neoformans var. grubii AFLP1/VNI Â C. gattii AFLP6/VGII (serotype AB, genotype AFLP11) (Meyer et al, 2009;Trilles et al, 2012;Cogliati et al, 2013;Kaocharoen et al, 2013;Hagen et al, 2015). These major molecular types have subsequently been confirmed using RFLP analysis of the orotidine monophosphate pyrophosphorylase (URA5) or phospholipase B (PLB1) genes, amplified fragment-length polymorphism (AFLP) and multi-locus sequence typing (MLST) analysis.…”

“…The URA5 gene was amplified using a specific pair of primers, URA5 and SJO1, as provided by Meyer et al (2009). The PCR mix comprised 1Â buffer (Promega), 1.5 mM MgCl 2 (Promega), 0.2 mM dNTPs (Sigma Aldrich), 0.2 µM each primer (Sigma Aldrich), 0.05 U Go Taq polymerase (Promega) µl À1 and 1 ng genomic DNA µl À1 .…”

“…Multiple molecular-typing methods have been described to study the epidemiology of the C. neoformans species complex. The most commonly used approaches to date involve AFLP, PCR fingerprinting and PCR-RFLP (Meyer et al, 2009;Illnait-Zaragozí et al, 2010b;Hagen et al, 2012). PCR fingerprinting and PCR-RFLP have proven useful in discriminating between the different serotypes and mating types, but have some limits for discrimination within specific C. neoformans species complex members.…”

“…The global incidence varies from 0.04 to 12 % and accounts for 13-40 % of the annual death rate among HIV-infected patients in areas of sub-Saharan Africa (Park et al, 2009). The fungal agent involved in the disease is the C. neoformans/Cryptococcus gattii species complex, belonging to the phylum Basidiomycota and subdivided into seven haploid and four hybrid genotypes (Trilles et al, 2008;Meyer et al, 2009;Cogliati et al, 2013;Hagen et al, 2015). C. neoformans var.…”

In vitro antifungal susceptibility profiles and genotypes of 308 clinical and environmental isolates of Cryptococcus neoformans var. grubii and Cryptococcus gattii serotype B from north-western India

“…grubii and C. gattii tested here were identified by standard mycological procedures (Randhawa et al, 2006(Randhawa et al, , 2008Hiremath et al, 2008). Their genotypes were determined based on two methods: (i) PCR fingerprinting using (GACA) 4 and M13 phage core sequences as single primers (Meyer et al, 1999); and (ii) DNA sequences at the URA5 locus (Meyer et al, 2009). The mating type of isolates was determined by PCR amplification, using primer pairs designed from the sequences of the mating type-specific STE12 and STE20 genes (Hiremath et al, 2008).…”