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.


CONCLUSIONS
PCR targeting the DNA topoisomerase II gene specifically and rapidly distinguished S. schenckii from its related species.
Kanbe
Toshio
T
Division of Molecular Mycology and Medicine, Department of Advanced Medical Science, Center for Neural Disease and Cancer, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya, Aichi 466-8550, Japan. tkanbe@med.nagoya-u.ac.jp
Natsume
Luna
L
Goto
Itsuko
I
Kawasaki
Masako
M
Mochizuki
Takashi
T
Ishizaki
Hiroshi
H
Kikuchi
Akihiko
A
D004247:DNA / Q000737:chemistry; D017931:DNA Primers / Q000737:chemistry; D004250:DNA Topoisomerases, Type II / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D005800:Genes, Fungal; D014644:Genetic Variation; D005838:Genotype; D005947:Glucose / Q000737:chemistry; D016533:Mycological Typing Techniques; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013175:Sporothrix / Q000235:genetics*
D017931:DNA Primers; D004271:DNA, Fungal; D004272:DNA, Mitochondrial; D004247:DNA; D004250:DNA Topoisomerases, Type II; D005947:Glucose