PCR-RFLP on β-tubulin gene for rapid identification of the most clinically important species of Aspergillus

Nasri, Tuba; Hedayati, Mohammad Taghi; Abastabar, Mahdi; Pasqualotto, Alessandro C.; Armaki, Mojtaba Taghizadeh; Hoseinnejad, Akbar; Nabili, Mojtaba

doi:10.1016/j.mimet.2015.08.007

Cited by 40 publications

(42 citation statements)

References 20 publications

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“…(n ϭ 2), Aspergillus uvarum (n ϭ 1), and Trichophyton tonsurans (n ϭ 1), were recovered from patients suffering from fingernail (n ϭ 70) and toenail (n ϭ 100) infections. Isolates were cultured on Sabouraud dextrose agar (Difco) supplemented with chloramphenicol for 2 to 7 days at 30°C and identified to the species level by PCR restriction fragment length polymorphism and DNA sequencing as previously described (23)(24)(25)(33)(34)(35)(36). In vitro antifungal susceptibility testing for filamentous fungi and yeast were performed according to Clinical and Laboratory Standards Institute (CLSI) documents M38-A2 and M27-A3, respectively (26,27).…”

mentioning

confidence: 99%

Low In Vitro Antifungal Activity of Tavaborole against Yeasts and Molds from Onychomycosis

Abastabar

Haghani

Shokohi

et al. 2018

Antimicrob Agents Chemother

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The activity of tavaborole, an FDA-approved antifungal drug, was compared to that of four antifungal agents against 170 clinical fungal isolates originating from patients with onychomycosis. Tavaborole had low activity against all isolates compared to itraconazole, terbinafine, and fluconazole, the principal choices for treatment of onychomycosis. Thus, it appears that tavaborole is not a candidate for the treatment of onychomycosis due to species, species, and dermatophytes.

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confidence: 99%

Low In Vitro Antifungal Activity of Tavaborole against Yeasts and Molds from Onychomycosis

Abastabar

Haghani

Shokohi

et al. 2018

Antimicrob Agents Chemother

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“…After centrifugation, the sediment was then divided into two parts; one for For molecular identification, genomic DNAs were extracted from all mold isolates, then universal primers including ITS1 and ITS4 as well as Bt2a and Bt2b (16) were used for identification of studied fungi to the species level. Herein, PCR reactions were prepared 17 and then the PCR products were analyzed by gel electrophoresis and visualized by UV illumination after safe staining. The amplified products of the ITS and β-tubulin fragments of isolated molds were conducted to automated DNA purification platform, and the purified amplicons were then sequenced.…”

Section: Sputum Processingmentioning

confidence: 99%

Prevalence of allergic bronchopulmonary aspergillosis in cystic fibrosis patients using two different diagnostic criteria

Maleki

Mortezaee

Hassanzad

et al. 2020

Eur Ann Allergy Clin Immunol

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Objective: There are different diagnostic criteria for the diagnosis of Allergic bronchopulmonary aspergillosis (ABPA) in CF patients. In this present study we evaluated the prevalence of ABPA in Iranian CF patients by two more usual diagnostic criteria as ISHAM working criteria (A) and CF Foundation Consensus Conference criteria (B). Methods: Eighty six CF patients were included in the study. All CF patients underwent for Aspergillus skin prick test (AST), Aspergillus-specific IgE (sIgEAf) and Aspergillus-specific IgG (sIgGAf), total IgE. The ABPA prevalence was estimated by two diagnostic criteria, (A) and (B) and compared.

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“…MG322299-322309) using two primers (Bt2a: 5¢-GGTAACCAAATCGGTGCTGCTTTC-3¢) and reverse (Bt2b: 5-ACCCTCAGTGTAGTGACCCTTGGC-3¢) as described previously. 21 Total DNA of A. clavatus strains was extracted from a hyphal plug taken from a fresh colony cultured on media using previously described procedures. 21,22 The coding sequences of the cyp51A and cyp51B genes were amplified for the isolates with low and high triazole MIC values using specific primer sets: Acla-A1 (5¢-CTGTATGGTTTGGGATTATAGG-3¢) and Acla-A2 (5¢-CCTCTGACACTTAGCTTAGG-3¢) for the cyp51A gene and Acla-B1 (5¢-CATGGTGCCTCTGTCTTATAG-3¢) and Acla-B2 (5¢-TCAGAAGGTCTCAAGGGTG-3¢) for the cyp51B gene.…”

Section: Pcr Amplification and Analysis Of Cyp51a And Cyp51b Gene Seqmentioning

confidence: 99%

“…21 Total DNA of A. clavatus strains was extracted from a hyphal plug taken from a fresh colony cultured on media using previously described procedures. 21,22 The coding sequences of the cyp51A and cyp51B genes were amplified for the isolates with low and high triazole MIC values using specific primer sets: Acla-A1 (5¢-CTGTATGGTTTGGGATTATAGG-3¢) and Acla-A2 (5¢-CCTCTGACACTTAGCTTAGG-3¢) for the cyp51A gene and Acla-B1 (5¢-CATGGTGCCTCTGTCTTATAG-3¢) and Acla-B2 (5¢-TCAGAAGGTCTCAAGGGTG-3¢) for the cyp51B gene. PCRs were carried out in 25 mL containing 2 mL genomic DNA, 1 mL of 25 pmol of each primer, 400 mM deoxynucleotide triphosphates, 2.5 U of Taq DNA polymerase, 2.5 mM MgCl 2 , and water.…”

Section: Pcr Amplification and Analysis Of Cyp51a And Cyp51b Gene Seqmentioning

confidence: 99%

Novel Point Mutations in cyp51A and cyp51B Genes Associated with Itraconazole and Posaconazole Resistance in Aspergillus clavatus Isolates

Abastabar

Hosseini

Valadan

et al. 2019

Microbial Drug Resistance

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Aspergillus clavatus is a common environmental species known to cause occupational allergic disease in grain handlers. We have recently observed azole-resistant isolates of this fungus as a cause of onychomycosis. To further characterize the cause of resistance, the genes encoding 14 a-sterol demethylase enzyme (cyp51A and cyp51B) were characterized and analyzed in 9 ITC-susceptible isolates and 6 isolates with high minimum inhibitory concentrations (MICs) of clinical (nail and sputum) and environmental A. clavatus strains. We found that six isolates with itraconazole MIC >16 mg/L demonstrated nonsynonymous mutations, including V51I, L378P, E483K, and E506G, and synonymous mutations, including F53F, A186A, Q276Q, and H359H. Moreover, P486S was detected in five strains with ITR MIC >16 mg/L. One mutation, F324S, was detected in an isolate with posaconazole MIC >16 mg/L. The effect of E483K and P486S mutations of CYP51A on azole resistance was further investigated using homology modeling and molecular dynamics. We found that E483K and P486S mutations were located near the ligand access channel of CYP51A that could partly lead to narrowing the entry of the ligand access channels. Therefore, we concluded that E483K and P486S mutations may potentially contribute to the limited access of inhibitors to the binding pocket and therefore confer resistance to azole agents.

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PCR-RFLP on β-tubulin gene for rapid identification of the most clinically important species of Aspergillus

Cited by 40 publications

References 20 publications

Low In Vitro Antifungal Activity of Tavaborole against Yeasts and Molds from Onychomycosis

Low In Vitro Antifungal Activity of Tavaborole against Yeasts and Molds from Onychomycosis

Prevalence of allergic bronchopulmonary aspergillosis in cystic fibrosis patients using two different diagnostic criteria

Novel Point Mutations in cyp51A and cyp51B Genes Associated with Itraconazole and Posaconazole Resistance in Aspergillus clavatus Isolates

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