Rapid identification of Sporothrix species by T3B fingerprinting

Abstract

This article describes PCR fingerprinting using the universal primer T3B to distinguish among species of the Sporothrix complex, S. brasiliensis, S. globosa, S. mexicana, and S. schenckii. This methodology generated distinct banding patterns, allowing the correct identification of all 35 clinical isolates at the species level, confirmed by partial calmodulin (CAL) gene sequence analyses. This methodology is simple, reliable, rapid, and cheap, making it an ideal routine identification system for clinical mycology laboratories.

Fig 1

Representative T3B PCR fingerprinting profiles of the Sporothrix complex. (1 and 7) Molecular marker DNA ladder, 100 bp (Invitrogen). (2) S. brasiliensis (IPEC 16490). (3) S. globosa (IPEC 27135). (4) S. mexicana (MUM 11.02). (5) S. schenckii (IPEC27722). (6) Negative control.

Fig 2

Dendrogram showing the degrees of similarity of T3B fingerprinting profiles among the Sporothrix isolates by using the Dice coefficient and the UPGMA cluster method. The cophenetic correlation coefficient (0.97) indicates a very good fit for this analysis.

Fig 3

Consensus tree of Sporothrix based on partial calmodulin (CAL) gene sequences of 35 strains and the NCBI public GenBank sequences AM398393.1 (S. mexicana), AM398382.1 (S. albicans), and AM117444.1 (S. schenckii) that was constructed with MEGA version 4.0.2 and 1,000 bootstrap replicates.