Molecular epidemiology of otomycosis in Isfahan revealed a large diversity in causative agents

Aboutalebian, Shima; Mahmoudi, Shahram; Mirhendi, Hossein; Okhovat, Ahmadreza; Abtahi, Hamidreza; Chabavizadeh, Javaher

doi:10.1099/jmm.0.000985

Cited by 27 publications

(37 citation statements)

References 25 publications

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“…A total of 120 patients clinically suspected of OE attending the otorhinolaryngology clinic of Al-Zahra Teaching Hospital, Isfahan, Iran, whose ear aspirations or ear swab specimens documented infective fungal and/or bacterial elements on direct microscopic examination by 10% Potassium Hydroxide and Giemsa mounts, were included in this study. For the identification of the yeasts isolated on CHROM agar Candida medium, the internal transcribed spacer (ITS) PCR-RFLP using the restriction enzymes MspI was performed as described in our previous report (Aboutalebian et al, 2019). Aspergillus/ Penicillium species which grew on the medium Sabouraud's dextrose agar supplemented with 50 mg L -1 chloramphenicol, were identified by partial b-tubulin gene sequencing.…”

Section: Patient and Isolatesmentioning

confidence: 99%

“…Aspergillus/ Penicillium species which grew on the medium Sabouraud's dextrose agar supplemented with 50 mg L -1 chloramphenicol, were identified by partial b-tubulin gene sequencing. Unidentified yeasts or molds (other than Aspergillus/ Penicillium) were subjected to the ITS sequencing (Aboutalebian et al, 2019). Furthermore, for the identification of bacterial agents, neither culture-based methods nor molecular procedures were applied; therefore, they were blindly investigated in the present study.…”

Section: Patient and Isolatesmentioning

confidence: 99%

“…Furthermore, for the identification of bacterial agents, neither culture-based methods nor molecular procedures were applied; therefore, they were blindly investigated in the present study. Written informed consent and detailed clinical data of the patient have been presented in our former report (Aboutalebian et al, 2019…”

Section: Patient and Isolatesmentioning

confidence: 99%

“…Pseudomonas aeruginosa and Staphylococcus aureus are frequently listed as the most predominant agents causing an acute form of OE (Schaefer and Baugh, 2012). On the other hand, fungal agents, most commonly Aspergillus and Candida species, are associated with prolonged antibiotic usage and chronic OE (Sarwestani et al, 2018;Aboutalebian et al, 2019;Punia et al, 2019). Complaints of acute onset of unilateral ear pain, itching, and a sense of aural fullness are the typical manifestations of acute otitis externa caused by bacteria.…”

Section: Introductionmentioning

confidence: 99%

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Direct Detection and Identification of the Most Common Bacteria and Fungi Causing Otitis Externa by a Stepwise Multiplex PCR

Aboutalebian

Ahmadikia

Fakhim

et al. 2021

Front. Cell. Infect. Microbiol.

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BackgroundConsidering the importance of differential diagnosis of infectious otitis externa (OE), a stepwise PCR-based assay using universal and genus- or species-specific primers for the detection/identification of the most prevalent bacterial and fungal OE was developed and evaluated on the ear aspiration specimens of clinically suspected patients.Methods and MaterialsA total of 120 ear aspiration specimens with otomycosis suspicion were subjected to manual DNA extraction using phenol–chloroform extraction after tissue digestion with a lysis buffer. The multiplex PCR was initially performed using pan-fungal and bacterial homemade primers. Pseudomonas and Staphylococcus specific primers were simultaneously used in one reaction mixture to identify the bacterial genera. Furthermore, for the identification of fungal agents, Candida species-specific multiplex primers targeting the most clinically important Candida species causing OE (i.e., C. albicans, C. parapsilosis, and C. auris), as well as Aspergillus related multiplex PCR identifying the most prevalent Aspergillus species were used in two separate reaction mixtures. All the results of multiplex PCR were interpreted based on the amplicon size.ResultsThe overall multiplex PCR-based detection rate of bacterial (n = 88; 73.3%) and fungal (n = 97; 81%) OE was documented to be 100% along with and complete consistency with the results of direct examination and Giemsa staining. Double amplicon bands of bacterial and fungal pathogens were evidenced in 76 specimens (63.3%). Moreover, the positivity rate of pan-fungal PCR was higher than that of the culture result. Out of 88 pan-bacterial positive PCR specimens, 66 and 47 ones were positive for Staphylococcus and Pseudomonas, respectively. In addition, 30 samples exhibited mixed infection of both, and five specimens remained negative. Out of 97 pan-fungal positive PCR specimens, 67 and 51 ones contained Candida and Aspergillus species, respectively. It should be noted that dual amplicon bands of Candida and Aspergillus-related multiplex PCR were yielded in 30 specimens.ConclusionThe stepwise multiplex PCR assay proved to be more sensitive, more rapid, as well as less cumbersome in detection and identification of fungal and bacterial OE, compared to culture.

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Section: Patient and Isolatesmentioning

confidence: 99%

Section: Patient and Isolatesmentioning

confidence: 99%

Section: Patient and Isolatesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Direct Detection and Identification of the Most Common Bacteria and Fungi Causing Otitis Externa by a Stepwise Multiplex PCR

Aboutalebian

Ahmadikia

Fakhim

et al. 2021

Front. Cell. Infect. Microbiol.

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“…Regarding the mold isolates, DNA was extracted by the phenol-chloroform method [19], and a fragment of ITS rDNA or beta-tubulin gene, depending on the genus of the isolate, was PCR-amplified and sent for sequencing as described previously [20,21]. Definitive identification of isolates was accomplished by the BLAST analysis (https://blast.ncbi.nlm.nih.gov/Blast.…”

Section: Identification Of Isolated Fungimentioning

confidence: 99%

Molecular characterization of fungi causing colonization and infection in organ transplant recipients: a one-year prospective study

Zarei¹,

Hashemi²,

Salehi³

et al. 2020

CMM

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Background and Purpose: Organ transplant recipients are vulnerable to fungal infections. The aim of this study was to determine the prevalence of fungal colonization and infections among patients who underwent various transplantations and molecularly characterize the etiological agents. Materials and Methods: This study was conducted on candidates for transplantation in Imam Khomeini Hospital, Tehran, Iran, from April 2017 to April 2018. All patients were monitored for fungal colonization or infections before and after transplantation. Isolated fungi were identified using molecular methods. Results: A total of 125 patients, including 86 males and 39 females, with the mean age of 52.2 years participated in the study (age range: 15-75 years). Out of 125 patients, 84 (67.2%) cases had fungal colonization that appeared pre- and post-transplantation in 21 and 63 cases, respectively (alone or concurrent with another infection in 55 and 29 cases, respectively). In addition, a total of 39 episodes of fungal infections were diagnosed in 36 (28.8%) recipients (alone or concurrent with colonization in 7 and 29 cases, respectively). Out of the 39 fungal infections, 9 cases appeared pre-transplantation, while the other 30 cases occurred post-transplantation. However, no fungal colonization or infection was observed in 34 (27.2%) patients. Oral candidiasis (n=20) was the most common type of infection, followed by funguria (n=7), onychomycosis (n=5), candidemia (n=3), rhinocerebral mucormycosis (n=1), cutaneous mucormycosis (n=1), cutaneous aspergillosis (n=1), and peritonitis (n=1). Six yeast species were recovered from colonization cases with the dominance of Candida albicans both before and after transplantation. The observed fungal infections were caused by 11 distinct species, including the members of Candida (i.e., C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, and C. krusei), Aspergillus (i.e., A. oryzae and A. candidus), Rhizopus (i.e., R. oryzae and R. microsporus), Trichosporon asahii, and Trichophyton interdigitale. The results also indicated that the development of a fungal infection post-transplantation was associated with fungal colonization (r=0.0184; P=0.043). Conclusion: Based on the results, fungal colonization was a common finding in transplant recipients at Imam Khomeini Hospital. However, the incidence of fungal infections was comparable with those of other centers. As the oral cavity was the most common site of colonization and infection, it might be beneficial to take further care about the oral health of patients using effective mouthwash.

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Combination of direct boiling and glass beads increases the purity and accuracy of bacterial DNA extraction

Liu

et al. 2023

Biotechnology Journal

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Extraction of DNA is a key step in molecular biology experiments and important for counting tiny microbial individuals. Direct boiling and mechanical cell lysis like glass beads are two independent physical extraction methods, thus crossing the barriers of thresholds of magnitude in popular reagent kits or traditional spread plate method when non‐equilibrium phenomenon is ongoing. The two approaches above were combined to generate a new one. In three typical microbial species, direct boiling with glass beads significantly increased the purity of DNA solution compared with some other methods (p < 0.05). The qPCR results of them were closer to direct microscopy counting than some other methods. Therefore, it provides a new choice in extracting bacterial DNA for specific circumstances.

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Molecular epidemiology of otomycosis in Isfahan revealed a large diversity in causative agents

Cited by 27 publications

References 25 publications

Direct Detection and Identification of the Most Common Bacteria and Fungi Causing Otitis Externa by a Stepwise Multiplex PCR

Direct Detection and Identification of the Most Common Bacteria and Fungi Causing Otitis Externa by a Stepwise Multiplex PCR

Molecular characterization of fungi causing colonization and infection in organ transplant recipients: a one-year prospective study

Combination of direct boiling and glass beads increases the purity and accuracy of bacterial DNA extraction

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