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

Shankarnarayan, Shamanth A.; Shaw, Dipika; Sharma, Anil; Chakrabarti, Arunaloke; Dogra, Sunil; Kumaran, Muthu Sendhil; Kaur, Harsimran; Ghosh, Anup; Rudramurthy, Shivaprakash M.

doi:10.1038/s41598-020-58187-0

Cited by 39 publications

(37 citation statements)

References 21 publications

(33 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Equally important is the elucidation of mechanisms used by dermatophytes to invade and colonise their hosts tissues as well as the signalling mechanisms and metabolic routes activated in response to antifungal drugs 15,16,32,48,52 . As suggested by many researchers, these points are essential to develop new and more effective therapeutic strategies 1,11,26,33,53‐55 . Most of the available articles discussing the mechanisms involved in dermatophyte resistance to antifungals are focused only im anthropophilic pathogens and are limited to such species as T. rubrum and T. interdigitale .…”

Section: Discussionmentioning

confidence: 99%

Complementary effect of mechanism of multidrug resistance in Trichophyton mentagrophytes isolated from human dermatophytoses of animal origin

Gnat

Łagowski

Nowakiewicz

et al. 2021

Mycoses

View full text Add to dashboard Cite

Background Dermatophytoses have gained interest worldwide due to the increased resistance to terbinafine and azoles and difficulty in management of these refractory diseases. Objectives In this study, we identified and analysed Trichophyton mentagrophytes clinical isolates obtained from humans with infections of animal origin. Methods We used quantitative real‐time PCR (qRT‐PCR) to examine the transcriptional modulation of three MDR genes (PDR1, MDR2 and MDR4) and analysed squalene epoxidase (SQLE) gene sequences from multidrug‐resistant Trichophyton mentagrophytes isolates. Results The expression profile revealed a 2‐ to 12‐fold increase in mRNA accumulation in the presence of any of the antifungals, compared to cells incubated without drugs. A statistically significant relationship between the isolates exposed to itraconazole and increased expression of the tested genes was revealed. Substantially lower transcription levels were noted for cells exposed to luliconazole, that is, a third‐generation azole. Additionally, in the case of 50% of terbinafine‐resistant strains, Leu397Phe substitution in the SQLE gene was detected. Furthermore, the reduced susceptibility to itraconazole and voriconazole was overcome by milbemycin oxime. Conclusions In conclusion, our study shed more light on the role of the ABC transporter family in T. mentagrophytes, which, if overexpressed, can confer resistance to single azole drugs and even cross‐resistance. Finally, milbemycin oxime could be an interesting compound supporting treatment with azole drugs in the case of refractory dermatomycoses.

show abstract

Section: Discussionmentioning

confidence: 99%

Complementary effect of mechanism of multidrug resistance in Trichophyton mentagrophytes isolated from human dermatophytoses of animal origin

Gnat

Łagowski

Nowakiewicz

et al. 2021

Mycoses

View full text Add to dashboard Cite

show abstract

“…The rapid detection of resistance to various antifungal drugs (azoles, echinocandins, and terbinafine) has been reported in many medically important fungi based on mutations in different resistance-related genes [ 30 , 35 – 38 ]. Resistance detection in many yeasts and molds has already been performed by using allele-specific real-time molecular probes, DNA microarray, HRM analysis, real-time PCR with molecular beacon probes, pyrosequencing, PCR-restriction fragment length polymorphism (PCR-RFLP), fluorescence resonance energy transfer (FRET), rolling circle amplification (RCA), ARMS-PCR, etc.…”

Section: Discussionmentioning

confidence: 99%

“…Resistance detection in many yeasts and molds has already been performed by using allele-specific real-time molecular probes, DNA microarray, HRM analysis, real-time PCR with molecular beacon probes, pyrosequencing, PCR-restriction fragment length polymorphism (PCR-RFLP), fluorescence resonance energy transfer (FRET), rolling circle amplification (RCA), ARMS-PCR, etc. [ 30 , 35 – 38 ]. Although studies have been performed on the rapid detection of ERG11 mutations associated with azole resistance in other Candida species, C .…”

Section: Discussionmentioning

confidence: 99%

“…T-ARMS-PCR is an efficient approach used for SNP genotyping [ 29 , 31 , 40 , 41 ]. A study from our clinical setup has been performed for the rapid terbinafine resistance detection in Trichophyton species by using conventional ARMS-PCR [ 30 ]. In conventional ARMS-PCR, the wild and mutant type alleles are amplified in two independent PCR reactions, in contrast, T-ARMS-PCR amplifies both alleles along with the control fragment together [ 29 , 31 ].…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Rapid detection of ERG11 polymorphism associated azole resistance in Candida tropicalis

et al. 2021

Self Cite

View full text Add to dashboard Cite

Increasing reports of azole resistance in Candida tropicalis, highlight the development of rapid resistance detection techniques. Nonsynonymous mutations in the lanosterol C14 alpha-demethylase (ERG11) gene is one of the predominant mechanisms of azole resistance in C. tropicalis. We evaluated the tetra primer-amplification refractory mutation system-PCR (T-ARMS-PCR), restriction site mutation (RSM), and high-resolution melt (HRM) analysis methods for rapid resistance detection based on ERG11 polymorphism in C. tropicalis. Twelve azole-resistant and 19 susceptible isolates of C. tropicalis were included. DNA sequencing of the isolates was performed to check the ERG11 polymorphism status among resistant and susceptible isolates. Three approaches T-ARMS-PCR, RSM, and HRM were evaluated and validated for the rapid detection of ERG11 mutation. The fluconazole MICs for the 12 resistant and 19 susceptible isolates were 32–256 mg/L and 0.5–1 mg/L, respectively. The resistant isolates showed A339T and C461T mutations in the ERG11 gene. The T-ARMS-PCR and RSM approaches discriminated all the resistant and susceptible isolates, whereas HRM analysis differentiated all except one susceptible isolate. The sensitivity, specificity, analytical sensitivity, time, and cost of analysis suggests that these three methods can be utilized for the rapid detection of ERG11 mutations in C. tropicalis. Additionally, an excellent concordance with DNA sequencing was noted for all three methods. The rapid, sensitive, and inexpensive T-ARMS-PCR, RSM, and HRM approaches are suitable for the detection of azole resistance based on ERG11 polymorphism in C. tropicalis and can be implemented in clinical setups for batter patient management.

show abstract

“…SNPs have been identified in squalene epoxidase homologs of Trichophyton group 19 . Nucleotide substitution within the squalene epoxidase gene was observed in Trichophyton interdigitale all isolates that substitutes TTC with CTC at 1185 position 20 . In Trichophyton rubrum, nucleotide substitution occurred at 1380, leading to the replacement of CAT with TAT 8 .…”

Section: Single Nucleotide Polymorphisms (Snps) In Squalene Epoxidase Genementioning

confidence: 98%

Metadata analysis of the squalene epoxidase gene in dermatophytes

Ismail¹,

Ahmad²,

Javed³

2021

Int. j. endorsing health sci. res.

View full text Add to dashboard Cite

Background: Squalene epoxidase gene is an azole antifungal target enzyme in the ergosterol biosynthesis pathway in fungi-the inhibition of enzyme aggregate squalene, a type of ergosterol derivative depletion that leads to fungal growth inhibition. Squalene epoxidase widely diverges in three fungal groups of dermatophytes Trichophyton, Microsporum and Epidermophyton. Methodology:The study has demonstrated a variety of squalene epoxidase genes in the dermatophyte genome. Squalene epoxidase gene was also studied for possible single nucleotide polymorphism (SNPs) in the Trichophyton group's homologs and found multiple SNP positions that induce non-synonymous mutation and change the sequence of amino acid into protein structure that can change phenylalanine to leucine. Results: Squalene epoxidase is widely present in dermatophytes. Mutation in the squalene epoxidase gene can mislead normal sterol formation in the fungal cell membrane that induces fungal resistance against several drugs, including azoles and polyenes. Squalene epoxidase gene carries 2 -3 transcripts and 2 -3 number of exons. Squalene epoxidase gene has FAD-dependent oxidoreductases and NADP binding domain conserved domains in fungal groups of dermatophytes. Conclusion:This study analyzed the abundance of the squalene epoxidase gene, suggesting that squalene epoxidase gene resistant mutants can occur naturally. Thus squalene epoxidase gene should be extensively studied in order to increase the potential of available antifungals.

show abstract

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

Cited by 39 publications

References 21 publications

Complementary effect of mechanism of multidrug resistance in Trichophyton mentagrophytes isolated from human dermatophytoses of animal origin

Complementary effect of mechanism of multidrug resistance in Trichophyton mentagrophytes isolated from human dermatophytoses of animal origin

Rapid detection of ERG11 polymorphism associated azole resistance in Candida tropicalis

Metadata analysis of the squalene epoxidase gene in dermatophytes

Contact Info

Product

Resources

About