Rapid and specific identification of Sporothrix schenckii by PCR targeting the DNA topoisomerase II gene.

BACKGROUND

In order to study the genotype variation of Sporothrix schenckii, we previously analyzed the genomic sequences of the DNA topoisomerase II genes of this fungus and S. schenckii-related species, such as S. schenckii var. luriei and Ceratocystis stenoceras.

OBJECTIVE

To develop a PCR-based identification system that can distinguish S. schenckii from its related species for clinical diagnosis of sporotrichosis.

METHODS

Based on the nucleotide sequences of the DNA topoisomerase II genes of S. schenckii, S. schenckii var. luriei and C. stenoceras, three gene-specific primers (SSHF31 and SSHR97 for S. schenckii, and SSLF64 for S. schenckii var. luriei) were designed and evaluated for their specificities in PCR amplifications.

RESULTS

The primer set SSHF31/SSHR97 amplified PCR products of 663-817 bp from S. schenckii and approximately 660 bp from S. schenckii var. luriei, while SSLF64/SSHR97 exclusively amplified a major product of 305 bp from S. schenckii var. luriei.

CONCLUSION

PCR targeting the DNA topoisomerase II gene specifically and rapidly distinguished S. schenckii from its related species.