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

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.

[1]  T. Jombart,et al.  Nonrandom Distribution of Azole Resistance across the Global Population of Aspergillus fumigatus , 2019, mBio.

[2]  T. Toyotome,et al.  A simple method to detect the tandem repeat of the cyp51A promoter in azole-resistant strains of Aspergillus fumigatus. , 2018, Medical mycology.

[3]  A. Chowdhary,et al.  Triazole resistance surveillance in Aspergillus fumigatus. , 2018, Medical mycology.

[4]  M. Bougnoux,et al.  Molecular Diagnosis of Invasive Aspergillosis and Detection of Azole Resistance by a Newly Commercialized PCR Kit , 2017, Journal of Clinical Microbiology.

[5]  D. Hagiwara,et al.  First clinical isolation report of azole-resistant Aspergillus fumigatus with TR34/L98H-type mutation in Japan. , 2017, Journal of infection and chemotherapy : official journal of the Japan Society of Chemotherapy.

[6]  W. Melchers,et al.  Genotype-phenotype complexity of the TR46/Y121F/T289A cyp51A azole resistance mechanism in Aspergillus fumigatus. , 2015, Fungal genetics and biology : FG & B.

[7]  P. White,et al.  Analytical and Clinical Evaluation of the PathoNostics AsperGenius Assay for Detection of Invasive Aspergillosis and Resistance to Azole Antifungal Drugs during Testing of Serum Samples , 2015, Journal of Clinical Microbiology.

[8]  G. Simons,et al.  Validation of a New Aspergillus Real-Time PCR Assay for Direct Detection of Aspergillus and Azole Resistance of Aspergillus fumigatus on Bronchoalveolar Lavage Fluid , 2015, Journal of Clinical Microbiology.

[9]  W. Melchers,et al.  The structure-function relationship of the Aspergillus fumigatuscyp51A L98H conversion by site-directed mutagenesis: the mechanism of L98H azole resistance. , 2011, Fungal genetics and biology : FG & B.

[10]  R. Saito,et al.  Rapid and Specific Detection of Amantadine-Resistant Influenza A Viruses with a Ser31Asn Mutation by the Cycling Probe Method , 2009, Journal of Clinical Microbiology.

[11]  Ken Noda,et al.  Rapid detection of quinolone-resistant Salmonella by real time SNP genotyping. , 2004, Journal of microbiological methods.

[12]  D. Hagiwara,et al.  Current Status of Azole-resistant Aspergillus fumigatus Isolates in East Asia. , 2018, Medical mycology journal.

[13]  D. Hagiwara,et al.  Azole-resistant Aspergillus fumigatus Containing a 34-bp Tandem Repeat in cyp51A Promoter is Isolated from the Environment in Japan. , 2017, Medical mycology journal.