For those North American isolates that are VGII by molecular type, the subtype-specific assays should be performed for typing VGIIa, VGIIb, or VGIIc. As we further our understanding of C. gattii populations around the world and their genotype-phenotype relationships, additional subtype specific assays can be similarly developed for local and global research purposes. Conclusions These PCR-based assays are an affordable,

efficient, and sensitive means of genotyping C. gattii isolates. Both the assay methods and results can be easily transferred among laboratories. Assay results are based on real-time PCR cycle threshold values and are therefore objective and straightforward for local analysis. The assay panel selleck kinase inhibitor presented here is a useful tool for conducting large-scale molecular epidemiological studies by public BVD-523 health and research laboratories. Ethics statement This study does not involve subjects or materials that would require approval by an ethics committee. Acknowledgements The findings and conclusions of this article are those of the authors and do not necessarily represent the views of the Centers for Disease Control and Prevention.

The authors wish to thank the members of the Cryptococcus gattii Public Health PD 332991 Working Group for submission of many of the isolates used in this study. This work was supported by funds from the National Institutes of Health: R21AI098059. References 1. Bovers M, Hagen F, Boekhout T: Diversity of the Cryptococcus neoformans-Cryptococcus gattii species complex. Rev Iberoam Micol 2008,25(1):S4-S12.PubMedCrossRef 2. D’Souza CA, Kronstad JW, Taylor G, Warren R, Yuen M, Hu G, Jung WH, Sham A, Kidd SE, Tangen K, Lee N, Zeilmaker T, Sawkins J, McVicker G, Shah S, Gnerre S, Griggs A, Zeng Q, Bartlett K, Li W, Wang X, Heitman J, Stajich JE, Fraser JA, Meyer

W, Carter D, Schein J, Krzywinski M, Kwon-Chung KJ, Varma A, et al.: Genome variation in Cryptococcus gattii , an emerging pathogen of immunocompetent hosts. MBio 2011, 2:e00342–10.PubMedCentralPubMed 3. Lockhart Protein Tyrosine Kinase inhibitor SR, Iqbal N, Bolden CB, DeBess EE, Marsden-Haug N, Worhle R, Thakur R, Harris JR: Epidemiologic cutoff values for triazole drugs in Cryptococcus gattii : correlation of molecular type and in vitro susceptibility. Diagn Microbiol Infect Dis 2012,73(2):144–148.PubMedCrossRef 4. Stephen CSL, Black W, Fyfe M, Raverty S: Multispecies outbreak of cryptococcosis on southern Vancouver Island, British Columbia. Can Vet J 2002,43(10):792–794.PubMedCentralPubMed 5. Iqbal N, DeBess EE, Wohrle R, Sun B, Nett RJ, Ahlquist AM, Chiller T, Lockhart SR: Correlation of genotype and in vitro susceptibilities of Cryptococcus gattii strains from the Pacific Northwest of the United States. J Clin Microbiol 2010,48(2):539–544.PubMedCentralPubMedCrossRef 6.