Rapid detection of terbinafine resistance in Trichophyton species by Amplified refractory mutation system-polymerase chain reaction

• Published in: Scientific reports Vol. 10; no. 1; p. 1297
• Main Authors: Shankarnarayan, Shamanth A., Shaw, Dipika, Sharma, Arunima, Chakrabarti, Arunaloke, Dogra, Sunil, Kumaran, Muthu Sendhil, Kaur, Harsimran, Ghosh, Anup, Rudramurthy, Shivaprakash M.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 28-01-2020; Nature Publishing Group
• Subjects: 45/22; 45/23; 45/29; 45/77; 631/326/193/2484; 692/4017; Amino acid substitution; Antifungal agents; Dermatomycosis; Disease resistance; DNA sequencing; Drug Resistance, Fungal - genetics; Fungal Proteins - genetics; Fungal Proteins - metabolism; Humanities and Social Sciences; Humans; multidisciplinary; Mutation; Point Mutation; Polymerase Chain Reaction; Science; Science (multidisciplinary); Single-nucleotide polymorphism; Species; Squalene; Squalene epoxidase; Squalene Monooxygenase - genetics; Squalene Monooxygenase - metabolism; Terbinafine; Tinea - drug therapy; Tinea - enzymology; Tinea - genetics; Transversion; Trichophyton - enzymology; Trichophyton - genetics
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-58187-0

Dermatophytosis has gained interest in India due to rise in terbinafine resistance and difficulty in management of recalcitrant disease. The terbinafine resistance in dermatophytes is attributed to single nucleotide polymorphisms (SNPs) in squalene epoxidase (
SE
) gene. We evaluated the utility of amplified refractory mutation system polymerase chain reaction (ARMS PCR) for detection of previously reported point mutations, including a mutation C1191A in the
SE
gene in
Trichophyton
species. ARMS PCR was standardized using nine non-wild type isolates and two wild type isolates of
Trichophyton
species. Study included 214 patients with dermatophyte infection from March through December 2017. Antifungal susceptibility testing of isolated dermatophytes was performed according to CLSI-M38A2 guidelines. Among dermatophytes isolated in 68.2% (146/214) patients,
Trichophyton
species were predominant (66.4%). High (>2 mg/L, cut off) minimum inhibitory concentrations to terbinafine were noted in 15 (15.4%)
Trichophyton mentagrophytes
complex isolates. A complete agreement was noted between ARMS PCR assay and DNA sequencing. C to A transversion was responsible for amino acid substitution in 397
th
position of
SE
gene in terbinafine resistant isolates. Thus, the ARMS PCR assay is a simple and reliable method to detect terbinafine-resistant
Trichophyton
isolates.