Article

New strain typing method with Sporothrix schenckii using mitochondrial DNA and polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) technique.

Department of Dermatology, School of Medicine, Kanazawa Medical University, Ishikawa, Japan.
The Journal of Dermatology (impact factor: 1.49). 09/2011; 39(4):362-5. DOI:10.1111/j.1346-8138.2011.01379.x pp.362-5
Source: PubMed

ABSTRACT The complete sequences of mitochondrial DNA (mtDNA) from two strains of different genotypes, American Type Culture Collection 10268 of mtDNA type 1 and KMU2025 of mtDNA type 4, were determined. These are circular molecules, 27 125 and 26 095 bp in length, respectively. The greatest difference between the two strains was found in the region encompassed by atp9 and cox2 genes, which was amplified with polymerase chain reaction (PCR) and used for preliminary restriction fragment length polymorphism (RFLP) analysis. Eight isolates of five mtDNA types were used and RFLP patterns obtained with the restriction enzyme AseI showed that this method seems to have greater discrimination power than the other PCR-RFLP typing method using internal transcribed spacer regions of nuclear DNA.

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Keywords

American Type Culture Collection 10268
 
atp9
 
complete sequences
 
different genotypes
 
greater discrimination power
 
greatest difference
 
internal transcribed spacer regions
 
mitochondrial DNA
 
mtDNA
 
mtDNA type 1
 
mtDNA type 4
 
mtDNA types
 
nuclear DNA
 
PCR-RFLP typing method
 
preliminary restriction fragment length polymorphism
 
restriction enzyme AseI