Detection of Substitutions at 98th, 121st, and 289th Amino Acid Residues in Cyp51A using Cycling Probes

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  • Toyotome Takahito
    Department of Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine Diagnostic Center for Animal Health and Food Safety, Obihiro University of Agriculture and Veterinary Medicine Division of Clinical Research, Medical Mycology Research Center, Chiba University
  • Arai Teppei
    Division of Clinical Research, Medical Mycology Research Center, Chiba University
  • Watanabe Akira
    Division of Clinical Research, Medical Mycology Research Center, Chiba University
  • Kamei Katsuhiko
    Division of Clinical Research, Medical Mycology Research Center, Chiba University

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Azole-resistant Aspergillus fumigatus containing unique mutation(s) of cyp51A with tandem repeats in the promoter region has emerged and has become dispersed in environments worldwide. For this study, we designed primers and cycling probes to detect mutations associated with tandem repeats. Substitutions at the 293rd nucleotide (leucine or histidine at the 98th amino acid residue), at the 362nd nucleotide (tyrosine or phenylalanine at the 121st amino acid residue), and at the 865th nucleotide (threonine or alanine at the 289th amino acid residue) in cyp51A were detected using these primers and probes. These results suggest that the primer and probe sets are helpful in detecting these mutations and in differentiating the types of tandem repeats in cyp51A.

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