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New strain typing method with Sporothrix schenckii using mitochondrial DNA and polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) technique
Authors:Kawasaki Masako  Anzawa Kazushi  Mochizuki Takashi  Ishizaki Hiroshi
Affiliation:Department of Dermatology, School of Medicine, Kanazawa Medical University, Ishikawa, Japan. masako-k@kanazawa-med.ac.jp
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.
Keywords:complete sequence of mitochondrion  mitochondrial DNA  polymerase chain reaction  restriction fragment length polymorphism  Sporothrix schenckii  strain typing method
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