Detection limit, specificity and the ability to detect H. capsulatum DNA in environmental samples with the MLST analysis. Lane M: marker. Sensitivity assay with H. capsulatum DNA (from lane 1 to 7): lane 1: 2 ng/μl, lane 2: 1 ng/μl, lane 3: 500 pg/μl, lane 4: 200 pg/μl, lane 5: 100 pg/μl, lane 6: 50 pg/μl, lane 7: 20 pg/μl. Specificity assay with DNA from different fungi (from lane 8 to 13): lane 8: Paracoccidioides brasiliensis sensu stricto (Pb 18), lane 9: Paracoccidioides spp. (Pb 339), lane 10: Paracoccidioides restrepiensis (Pb 60855), lane 11: Coccidioides spp, lane 12: Emmonsia crecens, lane 13: Blastomyces dermatitidis, lane 14: DNA from a raw chicken manure positive by Hc100 nested PCR and culture in Mycosel, lane 15: PCR negative control with sterile water. Non edited figure can be reviewed in the supplemental material (S4, S5 and S6 Figures).