Diagnosis and Molecular Characterization of Dermatophytosis: An Observational Study

Aditi Warghade; Gargi Mudey

doi:10.12688/f1000research.141657.2

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Study Protocol

Revised

Diagnosis and Molecular Characterization of Dermatophytosis: An Observational Study

[version 2; peer review: 1 approved, 1 not approved]

Previously titled: Study of molecular characterization for diagnosis of chronic and recurrent dermatophytosis

Aditi Warghade ¹, Gargi Mudey¹

PUBLISHED 08 Jul 2024

Author details Author details

¹ Department of Microbiology, Jawaharlal Nehru Medical College, Datta Meghe Institute of Higher Education and Research, Wardha, Maharashtra, 442001, India

Aditi Warghade
Roles: Conceptualization, Resources, Writing – Review & Editing

Gargi Mudey
Roles: Supervision, Validation, Writing – Original Draft Preparation

OPEN PEER REVIEW

REVIEWER STATUS

This article is included in the Datta Meghe Institute of Higher Education and Research collection.

Abstract

Dermatophytes are the keratinophilic fungi which infect humans and is the most recurring type of disease. The high level of transmissibility creates an epidemiological risk and emphasises the significance of these illnesses. However, a growing number of reports describing dermatophytes can cause deep infections in diabetic and immunocompromised patients, by invading deep layers like the dermis and hypodermis. Despite the prevalence and significance of dermatophytes in clinical mycology, it is not always possible to accurately diagnose this specific infection due to its overlapping structures among species of dermatophytes. Since it is difficult to identify species that exhibit weak characteristics in the morphological highlights, identification of the dermatophyte is often relied on its morphological analysis, which is a laborious process and demands skill. The massive shift in genetic variation, the source of infection, and epidemiological research can be discovered using molecular approaches. Therefore, the development of an accurate laboratory test for dermatophyte species identification is essential for the prevention and efficient management of dermatophytoses. One such methodology allows use of PCR technology which has many methods for molecular level characterization which is rapid, efficient, and capable of producing DNA polymorphisms specific to various dermatophyte species based on distinctive band patterns seen by agarose gel electrophoresis. The RAPD-PCR approach will be used in this study protocol to molecularly characterize the dermatophytes for precise speciation of the sample. In addition to improving knowledge of fungal biology and pathology with a focus on adaptive mechanisms to combat difficult conditions from host counteractions, there is a need to improve awareness of the importance of these diseases through accurate epidemiological data. The advantages of molecular approaches for characterizing objects over traditional methods are their sensitivity and specificity.

Keywords

Dermatophytosis, Molecular, Characterisation, RAPD-PCR, Electrophoresis, fungus, keratinophilic, Conventional

Corresponding authors: Aditi Warghade, Gargi Mudey

Competing interests: No competing interests were disclosed.

Grant information: An intramural grant from the Datta Meghe Institute of Higher Education and Research, Sawangi (Meghe), Wardha has been received.
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Copyright: © 2024 Warghade A and Mudey G. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

How to cite: Warghade A and Mudey G. Diagnosis and Molecular Characterization of Dermatophytosis: An Observational Study [version 2; peer review: 1 approved, 1 not approved]. F1000Research 2024, 13:136 (https://doi.org/10.12688/f1000research.141657.2) First published: 22 Feb 2024, 13:136 (https://doi.org/10.12688/f1000research.141657.1) Latest published: 08 Jul 2024, 13:136 (https://doi.org/10.12688/f1000research.141657.2)

Revised Amendments from Version 1

The article's title has been changed subject to some revisions suggested by the respected reviewers. Also recent changes in the T. rubrum to T. mentagrophytes that has occurred alongside the appearance of novel forms of tinea that are resistant to treatment. This dermatophyte is the primary cause of tinea cruris, tinea corporis, and tinea faciei in India, exceeding prior pathogens like T. rubrum. This information has been added with a reference of the same for further information. Also, study type and some new references have been added in the draft and cited as well.

See the authors' detailed response to the review by Saumya Panda
See the authors' detailed response to the review by Hassan Aboul-Ella

Introduction

Different clinical symptoms caused by dermatophytes range from superficial to subcutaneous affliction, including tinea corporis, tinea capitis, tinea pedis, and tinea unguium.¹ These diseases can range in severity clinically from moderate to severe depending on the host's immune system, the virulence of the strain, and other environmental factors.²^–⁴ Although dermatophytoses harm people throughout, they are more common in tropical regions due to high temperatures and humidity.⁵ Age, sex, time of year, socioeconomic and cultural circumstances, and geographic location are all factors that might influence the development of dermatophytosis.⁶ Potassium hydroxide (KOH) direct microscopy followed by selective medium culture is the standard procedure for dermatophyte screening in clinical samples. A quick screening technique for fungal structures is microscopy performed directly on clinical specimens, but this method lacks specificity.⁷ By using phenotypic techniques, dermatophyte isolates can be classified according to genus or species based on colony characteristics, microscopic assessments, and biochemical tests like growth patterns.⁸ Because there are changes from one isolate to another and overlaps in the features of several species, dermatophyte species identification by morphology in cultures may be difficult or imprecise.⁹ Molecular-based methods rely on identifying genotypic variations in pathogenic organisms.¹⁰^–¹² In India, a pathogen shift from T. rubrum to T. mentagrophytes has occurred alongside the appearance of novel forms of tinea that are resistant to treatment. This dermatophyte is the primary cause of tinea cruris, tinea corporis, and tinea faciei in India, exceeding prior pathogens like T. rubrum.¹³^,¹⁴ Molecular methods are being used to identify dermatophytes since they are more precise than conventional techniques.¹⁵ The development of molecular diagnostics were encouraged by the conventional techniques for identifying dermatophytes which are imprecise and have a delayed diagnostic character. Techniques that permit for both the early and accurate detection of dermatophytosis in order to provide timely antifungal treatment that prevents generic over-the-counter medication.¹⁶ Therefore, it is essential to create more reliable dermatophyte identification techniques.¹⁷ The current study's objective is to utilize RAPD-PCR method to identify and distinguish between the strains of fungi present in clinical isolates that cause chronic recurrent dermatophytosis.

Protocol

Study design

Oobservational study

Study participants: 100 participants having skin lesions, hair and nail positive for dermatophytic infection will be included in the study.

Study setting

• Samples (Skin scrapings, hair, nail clipping) will be collected from the patients visiting Dermatology OPD, Acharya Vinoba Bhave Rural Hospital, Sawangi (Meghe), Wardha.
• Samples will be processed in the Department of Microbiology, Jawaharlal Nehru Medical College, Sawangi (Meghe), Wardha.
• Method of selection of participants:
- 1. Patients having skin lesions will be included for taking the skin scrapings.
- 2. Patients with hair infections positive for fungal infection will be included for hair plucking.
- 3. Patients with nail infection will be considered for nail clippings.
All the samples will be collected in a black paper and stored in sterile container with proper labeling for further process.

Eligibility criteria

Patients having visible lesions and itching, positive for fungal infections are included in the study.

Data management

Samples will be collected directly from the patients and reports will be obtained from the microbiology laboratory. Data of all patients will be entered in Microsoft Excel taking proper precautions for wrapping the patient identifier information. The final deidentified data will be shared with statistician for further analysis.

Sample size

Sensitivity formula for calculating the sample size.

N \geq Z_{\underline{1 - α / 2}}^{\underline{2}} \frac{Sens (1 - Sens)}{d^{2} \times Pr ev}

Alpha (α) 0.05

Estimated sensitivity (Sens) 0.95

Prevalence of disease (Prev) 0.79¹⁸

Estimated error (d) 0.05

Minimum number of diseases needed: 73

Minimum total sample size needed: 93

Reagents required

1. Culture media Sabouraud Dextrose Agar (SDA)
2. Culture media Sabouraud Chloramphenicol + Cycloheximide Agar
3. Culture media Dermatophyte Agar Base
4. Dermato Supplement
5. Culture Media Corn meal agar
6. Distilled Water
7. Potassium Hydroxide Pellets
8. Normal Saline
9. 10% Glycerol
10. Lactophenol Cotton Blue Stain
11. Polyvinyl Alcohol
12. Ethyl Alcohol
13. Sodium Hypochlorite Solution
14. dNTP mix, 40mM
15. Hi-Temp PCR Master mix
16. Primers
17. AllPrep^® Fungal DNA isolation kit
18. PCR Block Plates
19. Pipette
20. Pipette tips (10μl, 20μl, 100μl, 1000μl)
21. Microscopic glass slides
22. Coverslip
23. Inoculating loop
24. Teasing needle
25. Petri dishes
26. Forceps
27. Glass Beakers
28. Flasks
29. Glass rod
30. Bunsen Burner
31. Gel electrophoresis chamber

Instruments required

1. Light Microscope
2. Autoclave
3. PCR
4. Biosafety Cabinet
5. Biological Oxygen Demand (BOD) Incubator
6. Electronic Weighing Machine

Protocol

A. Sample collection
- 1. The area of sample collection will be cleaned with 70% alcohol.
- 2. For skin samples, the lesion will be scraped around the corners using a scalpel blade or glass slide.
- 3. For nail samples, the nail clippings will be collected using a nail cutter in a sterile container
- 4. For hair samples, the hair will be plucked from the shaft of the hair having a lesion.
B. Sample processing
- 1. The skin scrapings will be dissolved in 10% KOH and nail clippings will be dissolved in 40% KOH for microscopic observation.
- 2. Samples will be inoculated on Dermatophyte Test Medium (DTM) as well as Sabouraud's dextrose agar (SDA) containing chloramphenicol and cycloheximide.
- 3. Samples will be incubated in BOD incubator, for observation of growth on the SDA and DTM slants.
- 4. Lactophenol cotton blue staining and slide culture technique will be used to view morphology and colony characteristics of the samples after growth.
C. Application of RAPD-PCR method for molecular characterization
- 1. Standardization of molecular assay will be done using the standard strains of Trichophyton (D15P127, CBS 118892, UCMS-IGIB-CI11), Microsporum (ATCC 36299), and laboratory-confirmed strains.
- 2. AllPrep® Fungal DNA isolation kit will be used for DNA isolation from fungal cultures.
- 3. The primer required for the RAPD-PCR reaction will be synthesized by Beacon designer probe/primer designer software from GeneX India Biosciences Velachery, Chennai, Tamil Nadu.
- 4. PCR assay mixture, reaction buffer, dNTPs, each primer set (GACA4) and novel primer (CTGT3), DNA template, using these PCR reaction cycles will be carried out.
  (39 cycles)
  - a) Denaturation at 93° - 1-minute,
  - b) Annealing step at 50° -1-minute,
  - c) Extension step at 72°- 1 minute
  - d) Final-extension step at 72°- 7 min
- 5. Final PCR products will be separated in 0.5X (Tris Borate-Ethylene diamine tetraacetic acid) Buffer and 1% Agarose and stained with the Ethidium-Bromide solution and then the image will be obtained.
- 6. Interspecies and intraspecies patterns and polymorphism for known strains will be studied.

Statistical analysis plan

All the results will be calculated using R version 4.3.2. Patients enrolled in the study sensitivity, specificity, positive predictive values (PPV), and negative predictive values (NPV) will be resulted from molecular methods used in comparison to the conventional method (gold standard).

Categorical variable will be summarized by the samples positive for fungal hyphae in KOH and culture on SDA (Sabouraud Dextrose Agar) and Dermatophyte Test Medium (DTM).

The percentage of agreement (overall, positive, and negative) between the two methods will be calculated using agreement analysis (primary and secondary endpoints), along with the Kappa coefficient, p-value, and 95% confidence interval.

Dissemination

Papers will be presented at relevant conferences and related studies will be published in indexed journals.

Study status

This research is ongoing. Using previously used primers, we are validating the protocol. The instruments, reagents, enzymes, and primers are set up. Clinical sample collection and processing of the sample by conventional method and standardization of molecular methods for the identification of species is ongoing.

Ethical considerations

This study has been granted by the ethics committee of Datta Meghe Institute of Higher Education and Research, Sawangi Meghe, Wardha with the approval number: DMIMS (DU)/IEC/2022/851, dated: 05/04/2022.

Written informed consent will be obtained from the study participants for participation in the study and publication of their data.

Discussion

Fungal infections in humans are becoming more common, especially in immunocompromised people, which has made them a global public health concern. The course of the disease, which can range from mild cutaneous or subcutaneous infections to invasive, widespread, and potentially fatal infections, is determined by the immunological health of the host.¹⁹ For epidemiological reasons, accurate antifungal treatment prevention of transmission includes exact separation between dermatophytosis and non-dermatophyte, and thorough identification of disease-causing organisms is vital.²⁰^,²¹ The most reliable approach to diagnose dermatophytosis is through the isolation and identification of dermatophytes from clinical samples. However, it typically takes a long time for the dermatophyte to grow in culture and sporulate, which delays diagnosis.²² Successful management of dermatophytes depends on prompt diagnosis and correct identification.²³ A study from Sweden by Ovrén, E et al. (2016) stated fluorescent staining method enhances the sensitivity and specificity in direct microscopy from skin, hair and nail samples and found that the specificity = (91.7–93.8%), positive predictive value (PPV) = (77.1–81.4%) and negative predictive value (NPV) = (83.7–89.9%).²⁴ Molecular methods are available for the characterization of the dermatophyte species namely restriction fragment length polymorphism (RFLP), (random amplified polymorphic DNA (RAPD), gene-specific-PCR,²⁵^,²⁶ chitin synthase encoding gene,²⁷ DNA hybridization,²⁸ and sequencing of the internal transcribed spacer region (ITS).⁷^,²⁹ A study by Li, H. C., Bouchara et al. (2007) from the United Kingdom stated the use of oligonucleotide array based on ITS-1 and ITS-2 sequence for identification of 17-dermatophyte species subsequently, hybridization of a series of oligonucleotides (17–30mers) digoxigenin-labeled PCR products immobilized on a nylon membrane of the 198 clinical dermal filamentous strains tested and 90 non-targeted strains, the array sensitivity and specificity were 99.5% and 97.8%, respectively.³⁰ Many molecular methods have made a way for early detection of dermatophytes to the species level.³¹ Along with these techniques serological methods have also detected the dermatophyte infection in the studies stated. A study by Higashi, Y et al. (2012) from Japan stated that the use of newly developed immunochromatographic strip-test for the diagnosis of dermatophytes found the overall sensitivity and specificity of the immunochromatographic test were 83.5% and 66.7%.³²^,³³

Data availability

No data are associated with this article.

Acknowledgments

We would like to acknowledge Department of Microbiology, JNMC and Central Research Laboratory (CRL) for their support.

References

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Comments on this article Comments (0)

Version 2

VERSION 2 PUBLISHED 22 Feb 2024

Author details Author details

¹ Department of Microbiology, Jawaharlal Nehru Medical College, Datta Meghe Institute of Higher Education and Research, Wardha, Maharashtra, 442001, India

Aditi Warghade
Roles: Conceptualization, Resources, Writing – Review & Editing

Gargi Mudey
Roles: Supervision, Validation, Writing – Original Draft Preparation

Competing interests

No competing interests were disclosed.

Grant information

An intramural grant from the Datta Meghe Institute of Higher Education and Research, Sawangi (Meghe), Wardha has been received.
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Article Versions (2)

version 2

Revised

Published: 08 Jul 2024, 13:136

https://doi.org/10.12688/f1000research.141657.2

version 1

Published: 22 Feb 2024, 13:136

https://doi.org/10.12688/f1000research.141657.1

© 2024 Warghade A and Mudey G. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Warghade A and Mudey G. Diagnosis and Molecular Characterization of Dermatophytosis: An Observational Study [version 2; peer review: 1 approved, 1 not approved]. F1000Research 2024, 13:136 (https://doi.org/10.12688/f1000research.141657.2)

NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article.

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Open Peer Review

Version 2

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PUBLISHED 08 Jul 2024

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Reviewer Report 22 Jul 2024

Saumya Panda, Jagannath Gupta Institute of Medical Sciences and Hospital, Kolkata, West Bengal, India

Not Approved

https://doi.org/10.5256/f1000research.168659.r300374

I thank the authors for their response. My comments on the current version are as follows:
1. While I appreciate the deletion of the characterization of dermatophytosis in the study as 'chronic and recurring', the subtitle "an observational study" is neither here nor there. While the earlier description of the study design as 'case-control' was plainly incorrect, the correct description would have been "a cross-sectional study". It is to be noted that 'case control' as well as 'cross-sectional' studies are all observational ones.
2. The reference given for the calculation of sample size is of a review article published in 2008. Other than being outdated, it is not possible to decipher exactly which Indian study has been taken as the prevalence reference.
3. The rationale of the study still remains inadequately addressed. One of the two references cited as explanation of the current epidemic was published in 1995!!!!

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Dermatology

I confirm that I have read this submission and believe that I have an appropriate level of expertise to state that I do not consider it to be of an acceptable scientific standard, for reasons outlined above.

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Reviewer Report 16 Jul 2024

Hassan Aboul-Ella, Cairo University, Giza, Giza Governorate, Egypt

Approved

https://doi.org/10.5256/f1000research.168659.r300373

The respected authors responded clearly to the previously addressed comments ... Continue reading

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Version 1

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PUBLISHED 22 Feb 2024

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Reviewer Report 14 May 2024

Saumya Panda, Jagannath Gupta Institute of Medical Sciences and Hospital, Kolkata, West Bengal, India

Approved with Reservations

https://doi.org/10.5256/f1000research.155121.r256151

This is an interesting and relevant protocol, given that there is an epidemic of chronic and recurrent dermatophytosis that is currently ongoing in our country as well as in several other countries and regions in the world. However, the protocol suffers from some serious lapses:
1. The protocol is titled as 'Study of molecular characterization for diagnosis of chronic and recurrent dermatophytosis'. However, the sampling methods and selection criteria of the cases do not mention these conditions at all!
2. Study design is mentioned as 'case control'. However, there is no description of the control group. The description matches that of a cross-sectional study, which is what it should be.
3. In sample size estimation, there is no reference to the prevalence value that has been taken, thus making the whole exercise infructuous.
4. The basic rationale of the study has not been mentioned - the current epidemic of recalcitrant dermatophytosis is due to the emergence of a new species called Trichophyton indotineae, which is morphologically and culturally indistinguishable from Trichohyton mentagrophytes and Trichophyton interdigitale. Thus its identification requires advanced molecular diagnostics like ITS genotyping or modified MALDI-TOF, which are not frequently available.

Is the rationale for, and objectives of, the study clearly described?

Partly
Is the study design appropriate for the research question?

No
Are sufficient details of the methods provided to allow replication by others?

Yes
Are the datasets clearly presented in a useable and accessible format?

Not applicable

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Dermatology

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above.

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Author Response 08 Jul 2024

Aditi Warghade, Department of Microbiology, Jawaharlal Nehru Medical College, Datta Meghe Institute of Higher Education and Research, Wardha, 442001, India

08 Jul 2024

Author Response

Respected Madam,
Thank you for considering our manuscript and suggesting the following changes to be processed for further publication in F1000 research subject to a satisfactory revision as per your ... Continue reading Respected Madam,
Thank you for considering our manuscript and suggesting the following changes to be processed for further publication in F1000 research subject to a satisfactory revision as per your suggestions. We have made the changes in our manuscript. Please consider the following changes.

This is an interesting and relevant protocol, given that there is an epidemic of chronic and recurrent dermatophytosis that is currently ongoing in our country as well as in several other countries and regions in the world. However, the protocol suffers from some serious lapses:
1. The protocol is titled as 'Study of molecular characterization for diagnosis of chronic and recurrent dermatophytosis'. However, the sampling methods and selection criteria of the cases do not mention these conditions at all!
Answer: Thank you for the suggestion mam. We have changed our title to "Diagnosis and Molecular Characterization of Dermatophytosis: An Observational Study". Track changes enabled and highlighted for the same.

2. Study design is mentioned as 'case-control'. However, there is no description of the control group. The description matches that of a cross-sectional study, which is what it should be.
Answer: Thank you for the suggestion, mam. We have stated the study design as an observational study as suggested.

3. In sample size estimation, there is no reference to the prevalence value taken, thus making the whole exercise infructuous.
Answer: Thank you for the suggestion mam. Mistakenly the study wasn’t added in the prevalence section. I have corrected the draft and added the study with Ref no-3 and highlighted the citation as it is not getting added in the reference list in sequence.

4. The basic rationale of the study has not been mentioned - the current epidemic of recalcitrant dermatophytosis is due to the emergence of a new species called Trichophyton indotineae, which is morphologically and culturally indistinguishable from Trichophyton mentagrophytes and Trichophyton interdigitale. Thus its identification requires advanced molecular diagnostics like ITS genotyping or modified MALDI-TOF, which are not frequently available.
Answer: Thank you for the suggestion, mam. I have added the reference information in the draft and quoted it as Ref no 1&2.
Respected Madam,
Thank you for considering our manuscript and suggesting the following changes to be processed for further publication in F1000 research subject to a satisfactory revision as per your suggestions. We have made the changes in our manuscript. Please consider the following changes.

This is an interesting and relevant protocol, given that there is an epidemic of chronic and recurrent dermatophytosis that is currently ongoing in our country as well as in several other countries and regions in the world. However, the protocol suffers from some serious lapses:
1. The protocol is titled as 'Study of molecular characterization for diagnosis of chronic and recurrent dermatophytosis'. However, the sampling methods and selection criteria of the cases do not mention these conditions at all!
Answer: Thank you for the suggestion mam. We have changed our title to "Diagnosis and Molecular Characterization of Dermatophytosis: An Observational Study". Track changes enabled and highlighted for the same.

2. Study design is mentioned as 'case-control'. However, there is no description of the control group. The description matches that of a cross-sectional study, which is what it should be.
Answer: Thank you for the suggestion, mam. We have stated the study design as an observational study as suggested.

3. In sample size estimation, there is no reference to the prevalence value taken, thus making the whole exercise infructuous.
Answer: Thank you for the suggestion mam. Mistakenly the study wasn’t added in the prevalence section. I have corrected the draft and added the study with Ref no-3 and highlighted the citation as it is not getting added in the reference list in sequence.

4. The basic rationale of the study has not been mentioned - the current epidemic of recalcitrant dermatophytosis is due to the emergence of a new species called Trichophyton indotineae, which is morphologically and culturally indistinguishable from Trichophyton mentagrophytes and Trichophyton interdigitale. Thus its identification requires advanced molecular diagnostics like ITS genotyping or modified MALDI-TOF, which are not frequently available.
Answer: Thank you for the suggestion, mam. I have added the reference information in the draft and quoted it as Ref no 1&2.
Competing Interests: Competing interests-none Close
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COMMENTS ON THIS REPORT

Author Response 08 Jul 2024

Aditi Warghade, Department of Microbiology, Jawaharlal Nehru Medical College, Datta Meghe Institute of Higher Education and Research, Wardha, 442001, India

08 Jul 2024

Author Response

Respected Madam,
Thank you for considering our manuscript and suggesting the following changes to be processed for further publication in F1000 research subject to a satisfactory revision as per your ... Continue reading Respected Madam,
Thank you for considering our manuscript and suggesting the following changes to be processed for further publication in F1000 research subject to a satisfactory revision as per your suggestions. We have made the changes in our manuscript. Please consider the following changes.

This is an interesting and relevant protocol, given that there is an epidemic of chronic and recurrent dermatophytosis that is currently ongoing in our country as well as in several other countries and regions in the world. However, the protocol suffers from some serious lapses:
1. The protocol is titled as 'Study of molecular characterization for diagnosis of chronic and recurrent dermatophytosis'. However, the sampling methods and selection criteria of the cases do not mention these conditions at all!
Answer: Thank you for the suggestion mam. We have changed our title to "Diagnosis and Molecular Characterization of Dermatophytosis: An Observational Study". Track changes enabled and highlighted for the same.

2. Study design is mentioned as 'case-control'. However, there is no description of the control group. The description matches that of a cross-sectional study, which is what it should be.
Answer: Thank you for the suggestion, mam. We have stated the study design as an observational study as suggested.

3. In sample size estimation, there is no reference to the prevalence value taken, thus making the whole exercise infructuous.
Answer: Thank you for the suggestion mam. Mistakenly the study wasn’t added in the prevalence section. I have corrected the draft and added the study with Ref no-3 and highlighted the citation as it is not getting added in the reference list in sequence.

4. The basic rationale of the study has not been mentioned - the current epidemic of recalcitrant dermatophytosis is due to the emergence of a new species called Trichophyton indotineae, which is morphologically and culturally indistinguishable from Trichophyton mentagrophytes and Trichophyton interdigitale. Thus its identification requires advanced molecular diagnostics like ITS genotyping or modified MALDI-TOF, which are not frequently available.
Answer: Thank you for the suggestion, mam. I have added the reference information in the draft and quoted it as Ref no 1&2.
Respected Madam,
Thank you for considering our manuscript and suggesting the following changes to be processed for further publication in F1000 research subject to a satisfactory revision as per your suggestions. We have made the changes in our manuscript. Please consider the following changes.

This is an interesting and relevant protocol, given that there is an epidemic of chronic and recurrent dermatophytosis that is currently ongoing in our country as well as in several other countries and regions in the world. However, the protocol suffers from some serious lapses:
1. The protocol is titled as 'Study of molecular characterization for diagnosis of chronic and recurrent dermatophytosis'. However, the sampling methods and selection criteria of the cases do not mention these conditions at all!
Answer: Thank you for the suggestion mam. We have changed our title to "Diagnosis and Molecular Characterization of Dermatophytosis: An Observational Study". Track changes enabled and highlighted for the same.

2. Study design is mentioned as 'case-control'. However, there is no description of the control group. The description matches that of a cross-sectional study, which is what it should be.
Answer: Thank you for the suggestion, mam. We have stated the study design as an observational study as suggested.

3. In sample size estimation, there is no reference to the prevalence value taken, thus making the whole exercise infructuous.
Answer: Thank you for the suggestion mam. Mistakenly the study wasn’t added in the prevalence section. I have corrected the draft and added the study with Ref no-3 and highlighted the citation as it is not getting added in the reference list in sequence.

4. The basic rationale of the study has not been mentioned - the current epidemic of recalcitrant dermatophytosis is due to the emergence of a new species called Trichophyton indotineae, which is morphologically and culturally indistinguishable from Trichophyton mentagrophytes and Trichophyton interdigitale. Thus its identification requires advanced molecular diagnostics like ITS genotyping or modified MALDI-TOF, which are not frequently available.
Answer: Thank you for the suggestion, mam. I have added the reference information in the draft and quoted it as Ref no 1&2.
Competing Interests: Competing interests-none Close
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Views

Reviewer Report 27 Mar 2024

Hassan Aboul-Ella, Cairo University, Giza, Giza Governorate, Egypt

Approved with Reservations

https://doi.org/10.5256/f1000research.155121.r251787

The current work describes a protocol for an observational study through collecting samples from dermatophytes-suffering patients followed by conventional and molecular diagnostic process, the obtained data will be very useful in improving and updating the epidemiological data in this field.

General comments

While the research focuses on an encouraging area of research and addresses an area of study with minimal existing research, it requires substantial editing of vocabulary in English especially in the following sections; abstract, introduction, and discussion.

Specific comments

- The title needs certain modifications to be more representable for the designed study. Therefore, I suggest "Diagnosis and molecular characterization of dermatophytosis: an observational study"

- The abstract, introduction, and methodological protocol sections are well designed, structured, and described.

- The discussion is missing certain necessary references to support the flow of the work and the researcher point of view in the following points;

1) The most reliable approach to diagnose dermatophytosis is through the isolation and identification of dermatophytes from clinical samples. However, it typically takes a long time for the dermatophyte to grow in culture and sporulate, which delays diagnosis.
- The suggested [Ref 2] in which a culture-dependent cross sectional study has been performed on dermatophytosis showing the importance of culture in dermatophytosis diagnosis as well as the extend of to which extend this technique is laborious and time consuming and that will support the researcher point of view.

2)  Many molecular methods have made a way for early detection of dermatophytes to the species level.
- The suggested [Ref 3] in which a comprehensive and collective mentioning and illustration of the molecular advancement and molecular diagnostic techniques that is have been developed in the field of dermatophytosis and the extend of the diagnostic level of those techniques and that of course will strongly support the researcher point of view.

3) A study by Higashi, Y et al. (2012) from Japan stated that the use of newly developed immunochromatographic strip-test for the diagnosis of dermatophytes found the overall sensitivity and specificity of the immunochromatographic test were 83.5% and 66.7%.
- The suggested [Ref 1] with all respect of the cited study as Higashi et al. 2012, study in the field of immunochromatographic diagnosis of dermatophytosis is really a pioneering study in this field but to be more accurate and more updated I suggest adding the suggested reference in addition to the Higashi et al. 2012, study as the suggested reference represents a newest designed and developed immunochromatographic assay for dermatophytosis diagnosis with a competitive results and certain important production modification over that of Higashi kits.

Is the rationale for, and objectives of, the study clearly described?

Yes
Is the study design appropriate for the research question?

Yes
Are sufficient details of the methods provided to allow replication by others?

Yes
Are the datasets clearly presented in a useable and accessible format?

Not applicable

References

1. Aboul-Ella H, Sayed RH, Abo-Elyazeed HS: Development, preparation, and evaluation of a novel dotted lateral flow immunochromatographic kit for rapid diagnosis of dermatophytosis.Sci Rep. 2023; 13 (1): 248 PubMed Abstract | Publisher Full Text
2. A. Hassanien H, H. Sayed R, S. Abo Elyazeed H: Cross-Sectional Study on Dermatological Affections of Companion Animals Caused by Dermatophytes and other Keratinophilic Fungi in Greater Cairo Area, Egypt. Advances in Animal and Veterinary Sciences. 2020; 9 (4). Publisher Full Text
3. Aboul-Ella H, Hamed R, Abo-Elyazeed H: Recent trends in rapid diagnostic techniques for dermatophytosis.Int J Vet Sci Med. 2020; 8 (1): 115-123 PubMed Abstract | Publisher Full Text

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Mycology, dermatophytosis, and mycological diagnostic techniques are my major research areas

CITE

Report a concern

Author Response 08 Jul 2024

Aditi Warghade, Department of Microbiology, Jawaharlal Nehru Medical College, Datta Meghe Institute of Higher Education and Research, Wardha, 442001, India

08 Jul 2024

Author Response

Respected sir,
Thank you for considering our manuscript and suggesting the following changes to be processed for further publication in F1000 research subject to a satisfactory revision as per your ... Continue reading Respected sir,
Thank you for considering our manuscript and suggesting the following changes to be processed for further publication in F1000 research subject to a satisfactory revision as per your suggestions. We have made the changes in our manuscript as per the suggestions given. Please consider the following changes

The title needs certain modifications to be more representable for the designed study. Therefore, I suggest "Diagnosis and molecular characterization of dermatophytosis: an observational study"
Answer:
Thank you for the suggestion, sir. I agree with the changes suggested and would like to change the title to "Diagnosis and Molecular Characterization of Dermatophytosis: An Observational Study"

- The abstract, introduction, and methodological protocol sections are well-designed, structured, and described.
- The discussion is missing certain necessary references to support the flow of the work and the researcher's point of view in the following points;

1) The most reliable approach to diagnose dermatophytosis is through the isolation and identification of dermatophytes from clinical samples. However, it typically takes a long time for the dermatophyte to grow in culture and sporulate, which delays diagnosis.
- The suggested [Ref 2] in which a culture-dependent cross-sectional study has been performed on dermatophytosis showing the importance of culture in dermatophytosis diagnosis as well as the extend of to which extend this technique is laborious and time consuming and that will support the researcher point of view.
Answer: Thank you for the comment, sir. We have added the suggested reference in the draft, quoted as Ref No-4, and highlighted the citation as it is not getting added in the reference list in sequence.

2)  Many molecular methods have made a way for early detection of dermatophytes to the species level.
- The suggested [Ref 3] in which a comprehensive and collective mentioning and illustration of the molecular advancement and molecular diagnostic techniques that is have been developed in the field of dermatophytosis and the extend of the diagnostic level of those techniques and that of course will strongly support the researcher point of view.
Answer: Thank you for the suggestion, sir. We have added the suggested reference in the draft quoted as Ref No- 5, and highlighted the citation as it is not getting added in the reference list in sequence.

3) A study by Higashi, Y et al. (2012) from Japan stated that the use of newly developed immunochromatographic strip-test for the diagnosis of dermatophytes found the overall sensitivity and specificity of the immunochromatographic test were 83.5% and 66.7%.
- The suggested [Ref 1] with all respect of the cited study as Higashi et al. 2012, study in the field of immunochromatographic diagnosis of dermatophytosis is really a pioneering study in this field but to be more accurate and more updated I suggest adding the suggested reference in addition to the Higashi et al. 2012, study as the suggested reference represents a newest designed and developed immunochromatographic assay for dermatophytosis diagnosis with a competitive results and certain important production modification over that of Higashi kits..
Answer: Thank you for the suggestion, sir. We have added the suggested reference in our study and quoted it as Ref No-6, and highlighted the citation as it is not getting added in the reference list in sequence.
Respected sir,
Thank you for considering our manuscript and suggesting the following changes to be processed for further publication in F1000 research subject to a satisfactory revision as per your suggestions. We have made the changes in our manuscript as per the suggestions given. Please consider the following changes

The title needs certain modifications to be more representable for the designed study. Therefore, I suggest "Diagnosis and molecular characterization of dermatophytosis: an observational study"
Answer:
Thank you for the suggestion, sir. I agree with the changes suggested and would like to change the title to "Diagnosis and Molecular Characterization of Dermatophytosis: An Observational Study"

- The abstract, introduction, and methodological protocol sections are well-designed, structured, and described.
- The discussion is missing certain necessary references to support the flow of the work and the researcher's point of view in the following points;

1) The most reliable approach to diagnose dermatophytosis is through the isolation and identification of dermatophytes from clinical samples. However, it typically takes a long time for the dermatophyte to grow in culture and sporulate, which delays diagnosis.
- The suggested [Ref 2] in which a culture-dependent cross-sectional study has been performed on dermatophytosis showing the importance of culture in dermatophytosis diagnosis as well as the extend of to which extend this technique is laborious and time consuming and that will support the researcher point of view.
Answer: Thank you for the comment, sir. We have added the suggested reference in the draft, quoted as Ref No-4, and highlighted the citation as it is not getting added in the reference list in sequence.

2)  Many molecular methods have made a way for early detection of dermatophytes to the species level.
- The suggested [Ref 3] in which a comprehensive and collective mentioning and illustration of the molecular advancement and molecular diagnostic techniques that is have been developed in the field of dermatophytosis and the extend of the diagnostic level of those techniques and that of course will strongly support the researcher point of view.
Answer: Thank you for the suggestion, sir. We have added the suggested reference in the draft quoted as Ref No- 5, and highlighted the citation as it is not getting added in the reference list in sequence.

3) A study by Higashi, Y et al. (2012) from Japan stated that the use of newly developed immunochromatographic strip-test for the diagnosis of dermatophytes found the overall sensitivity and specificity of the immunochromatographic test were 83.5% and 66.7%.
- The suggested [Ref 1] with all respect of the cited study as Higashi et al. 2012, study in the field of immunochromatographic diagnosis of dermatophytosis is really a pioneering study in this field but to be more accurate and more updated I suggest adding the suggested reference in addition to the Higashi et al. 2012, study as the suggested reference represents a newest designed and developed immunochromatographic assay for dermatophytosis diagnosis with a competitive results and certain important production modification over that of Higashi kits..
Answer: Thank you for the suggestion, sir. We have added the suggested reference in our study and quoted it as Ref No-6, and highlighted the citation as it is not getting added in the reference list in sequence.
Competing Interests: Competing interests-none Close
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Respond or Comment

COMMENTS ON THIS REPORT

Author Response 08 Jul 2024

Aditi Warghade, Department of Microbiology, Jawaharlal Nehru Medical College, Datta Meghe Institute of Higher Education and Research, Wardha, 442001, India

08 Jul 2024

Author Response

Respected sir,
Thank you for considering our manuscript and suggesting the following changes to be processed for further publication in F1000 research subject to a satisfactory revision as per your ... Continue reading Respected sir,
Thank you for considering our manuscript and suggesting the following changes to be processed for further publication in F1000 research subject to a satisfactory revision as per your suggestions. We have made the changes in our manuscript as per the suggestions given. Please consider the following changes

The title needs certain modifications to be more representable for the designed study. Therefore, I suggest "Diagnosis and molecular characterization of dermatophytosis: an observational study"
Answer:
Thank you for the suggestion, sir. I agree with the changes suggested and would like to change the title to "Diagnosis and Molecular Characterization of Dermatophytosis: An Observational Study"

- The abstract, introduction, and methodological protocol sections are well-designed, structured, and described.
- The discussion is missing certain necessary references to support the flow of the work and the researcher's point of view in the following points;

1) The most reliable approach to diagnose dermatophytosis is through the isolation and identification of dermatophytes from clinical samples. However, it typically takes a long time for the dermatophyte to grow in culture and sporulate, which delays diagnosis.
- The suggested [Ref 2] in which a culture-dependent cross-sectional study has been performed on dermatophytosis showing the importance of culture in dermatophytosis diagnosis as well as the extend of to which extend this technique is laborious and time consuming and that will support the researcher point of view.
Answer: Thank you for the comment, sir. We have added the suggested reference in the draft, quoted as Ref No-4, and highlighted the citation as it is not getting added in the reference list in sequence.

2)  Many molecular methods have made a way for early detection of dermatophytes to the species level.
- The suggested [Ref 3] in which a comprehensive and collective mentioning and illustration of the molecular advancement and molecular diagnostic techniques that is have been developed in the field of dermatophytosis and the extend of the diagnostic level of those techniques and that of course will strongly support the researcher point of view.
Answer: Thank you for the suggestion, sir. We have added the suggested reference in the draft quoted as Ref No- 5, and highlighted the citation as it is not getting added in the reference list in sequence.

3) A study by Higashi, Y et al. (2012) from Japan stated that the use of newly developed immunochromatographic strip-test for the diagnosis of dermatophytes found the overall sensitivity and specificity of the immunochromatographic test were 83.5% and 66.7%.
- The suggested [Ref 1] with all respect of the cited study as Higashi et al. 2012, study in the field of immunochromatographic diagnosis of dermatophytosis is really a pioneering study in this field but to be more accurate and more updated I suggest adding the suggested reference in addition to the Higashi et al. 2012, study as the suggested reference represents a newest designed and developed immunochromatographic assay for dermatophytosis diagnosis with a competitive results and certain important production modification over that of Higashi kits..
Answer: Thank you for the suggestion, sir. We have added the suggested reference in our study and quoted it as Ref No-6, and highlighted the citation as it is not getting added in the reference list in sequence.
Respected sir,
Thank you for considering our manuscript and suggesting the following changes to be processed for further publication in F1000 research subject to a satisfactory revision as per your suggestions. We have made the changes in our manuscript as per the suggestions given. Please consider the following changes

The title needs certain modifications to be more representable for the designed study. Therefore, I suggest "Diagnosis and molecular characterization of dermatophytosis: an observational study"
Answer:
Thank you for the suggestion, sir. I agree with the changes suggested and would like to change the title to "Diagnosis and Molecular Characterization of Dermatophytosis: An Observational Study"

- The abstract, introduction, and methodological protocol sections are well-designed, structured, and described.
- The discussion is missing certain necessary references to support the flow of the work and the researcher's point of view in the following points;

1) The most reliable approach to diagnose dermatophytosis is through the isolation and identification of dermatophytes from clinical samples. However, it typically takes a long time for the dermatophyte to grow in culture and sporulate, which delays diagnosis.
- The suggested [Ref 2] in which a culture-dependent cross-sectional study has been performed on dermatophytosis showing the importance of culture in dermatophytosis diagnosis as well as the extend of to which extend this technique is laborious and time consuming and that will support the researcher point of view.
Answer: Thank you for the comment, sir. We have added the suggested reference in the draft, quoted as Ref No-4, and highlighted the citation as it is not getting added in the reference list in sequence.

2)  Many molecular methods have made a way for early detection of dermatophytes to the species level.
- The suggested [Ref 3] in which a comprehensive and collective mentioning and illustration of the molecular advancement and molecular diagnostic techniques that is have been developed in the field of dermatophytosis and the extend of the diagnostic level of those techniques and that of course will strongly support the researcher point of view.
Answer: Thank you for the suggestion, sir. We have added the suggested reference in the draft quoted as Ref No- 5, and highlighted the citation as it is not getting added in the reference list in sequence.

3) A study by Higashi, Y et al. (2012) from Japan stated that the use of newly developed immunochromatographic strip-test for the diagnosis of dermatophytes found the overall sensitivity and specificity of the immunochromatographic test were 83.5% and 66.7%.
- The suggested [Ref 1] with all respect of the cited study as Higashi et al. 2012, study in the field of immunochromatographic diagnosis of dermatophytosis is really a pioneering study in this field but to be more accurate and more updated I suggest adding the suggested reference in addition to the Higashi et al. 2012, study as the suggested reference represents a newest designed and developed immunochromatographic assay for dermatophytosis diagnosis with a competitive results and certain important production modification over that of Higashi kits..
Answer: Thank you for the suggestion, sir. We have added the suggested reference in our study and quoted it as Ref No-6, and highlighted the citation as it is not getting added in the reference list in sequence.
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VERSION 2 PUBLISHED 22 Feb 2024

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Hassan Aboul-Ella, Cairo University, Giza, Egypt
Saumya Panda, Jagannath Gupta Institute of Medical Sciences and Hospital, Kolkata, India

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22 Jul 2024 | for Version 2

Saumya Panda, Jagannath Gupta Institute of Medical Sciences and Hospital, Kolkata, West Bengal, India

7 Views Cite this report Responses(0)

Not Approved

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Dermatology

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4 Views

16 Jul 2024 | for Version 2

Hassan Aboul-Ella, Cairo University, Giza, Giza Governorate, Egypt

4 Views Cite this report Responses(0)

Approved

The respected authors responded clearly to the previously addressed comments and I don't have any further comments at this stage.

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Mycology, dermatophytosis, and mycological diagnostic techniques are my major research areas

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.

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17 Views

14 May 2024 | for Version 1

Saumya Panda, Jagannath Gupta Institute of Medical Sciences and Hospital, Kolkata, West Bengal, India

17 Views Cite this report Responses(1)

Approved With Reservations

Is the rationale for, and objectives of, the study clearly described?

Partly
Is the study design appropriate for the research question?

No
Are sufficient details of the methods provided to allow replication by others?

Yes
Are the datasets clearly presented in a useable and accessible format?

Not applicable

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Dermatology

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Author Response

08 Jul 2024

Aditi Warghade, Department of Microbiology, Jawaharlal Nehru Medical College, Datta Meghe Institute of Higher Education and Research, Wardha, 442001, India

Respected Madam,
Thank you for considering our manuscript and suggesting the following changes to be processed for further publication in F1000 research subject to a satisfactory revision as per your suggestions. We have made the changes in our manuscript. Please consider the following changes.

This is an interesting and relevant protocol, given that there is an epidemic of chronic and recurrent dermatophytosis that is currently ongoing in our country as well as in several other countries and regions in the world. However, the protocol suffers from some serious lapses:
1. The protocol is titled as 'Study of molecular characterization for diagnosis of chronic and recurrent dermatophytosis'. However, the sampling methods and selection criteria of the cases do not mention these conditions at all!
Answer: Thank you for the suggestion mam. We have changed our title to "Diagnosis and Molecular Characterization of Dermatophytosis: An Observational Study". Track changes enabled and highlighted for the same.

2. Study design is mentioned as 'case-control'. However, there is no description of the control group. The description matches that of a cross-sectional study, which is what it should be.
Answer: Thank you for the suggestion, mam. We have stated the study design as an observational study as suggested.

3. In sample size estimation, there is no reference to the prevalence value taken, thus making the whole exercise infructuous.
Answer: Thank you for the suggestion mam. Mistakenly the study wasn’t added in the prevalence section. I have corrected the draft and added the study with Ref no-3 and highlighted the citation as it is not getting added in the reference list in sequence.

4. The basic rationale of the study has not been mentioned - the current epidemic of recalcitrant dermatophytosis is due to the emergence of a new species called Trichophyton indotineae, which is morphologically and culturally indistinguishable from Trichophyton mentagrophytes and Trichophyton interdigitale. Thus its identification requires advanced molecular diagnostics like ITS genotyping or modified MALDI-TOF, which are not frequently available.
Answer: Thank you for the suggestion, mam. I have added the reference information in the draft and quoted it as Ref no 1&2.

View more View less

Competing Interests

Competing interests-none

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Reviewer Report

30 Views

27 Mar 2024 | for Version 1

Hassan Aboul-Ella, Cairo University, Giza, Giza Governorate, Egypt

30 Views Cite this report Responses(1)

Approved With Reservations

Is the rationale for, and objectives of, the study clearly described?

Yes
Is the study design appropriate for the research question?

Yes
Are sufficient details of the methods provided to allow replication by others?

Yes
Are the datasets clearly presented in a useable and accessible format?

Not applicable

References

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Mycology, dermatophytosis, and mycological diagnostic techniques are my major research areas

Respond to this report

Responses (1)

Author Response

08 Jul 2024

Aditi Warghade, Department of Microbiology, Jawaharlal Nehru Medical College, Datta Meghe Institute of Higher Education and Research, Wardha, 442001, India

Respected sir,
Thank you for considering our manuscript and suggesting the following changes to be processed for further publication in F1000 research subject to a satisfactory revision as per your suggestions. We have made the changes in our manuscript as per the suggestions given. Please consider the following changes

The title needs certain modifications to be more representable for the designed study. Therefore, I suggest "Diagnosis and molecular characterization of dermatophytosis: an observational study"
Answer:
Thank you for the suggestion, sir. I agree with the changes suggested and would like to change the title to "Diagnosis and Molecular Characterization of Dermatophytosis: An Observational Study"

- The abstract, introduction, and methodological protocol sections are well-designed, structured, and described.
- The discussion is missing certain necessary references to support the flow of the work and the researcher's point of view in the following points;

1) The most reliable approach to diagnose dermatophytosis is through the isolation and identification of dermatophytes from clinical samples. However, it typically takes a long time for the dermatophyte to grow in culture and sporulate, which delays diagnosis.
- The suggested [Ref 2] in which a culture-dependent cross-sectional study has been performed on dermatophytosis showing the importance of culture in dermatophytosis diagnosis as well as the extend of to which extend this technique is laborious and time consuming and that will support the researcher point of view.
Answer: Thank you for the comment, sir. We have added the suggested reference in the draft, quoted as Ref No-4, and highlighted the citation as it is not getting added in the reference list in sequence.

2) Many molecular methods have made a way for early detection of dermatophytes to the species level.
- The suggested [Ref 3] in which a comprehensive and collective mentioning and illustration of the molecular advancement and molecular diagnostic techniques that is have been developed in the field of dermatophytosis and the extend of the diagnostic level of those techniques and that of course will strongly support the researcher point of view.
Answer: Thank you for the suggestion, sir. We have added the suggested reference in the draft quoted as Ref No- 5, and highlighted the citation as it is not getting added in the reference list in sequence.

3) A study by Higashi, Y et al. (2012) from Japan stated that the use of newly developed immunochromatographic strip-test for the diagnosis of dermatophytes found the overall sensitivity and specificity of the immunochromatographic test were 83.5% and 66.7%.
- The suggested [Ref 1] with all respect of the cited study as Higashi et al. 2012, study in the field of immunochromatographic diagnosis of dermatophytosis is really a pioneering study in this field but to be more accurate and more updated I suggest adding the suggested reference in addition to the Higashi et al. 2012, study as the suggested reference represents a newest designed and developed immunochromatographic assay for dermatophytosis diagnosis with a competitive results and certain important production modification over that of Higashi kits..
Answer: Thank you for the suggestion, sir. We have added the suggested reference in our study and quoted it as Ref No-6, and highlighted the citation as it is not getting added in the reference list in sequence.

View more View less

Competing Interests

Competing interests-none

Alongside their report, reviewers assign a status to the article:

Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested

Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit.

Not approved - fundamental flaws in the paper seriously undermine the findings and conclusions

[1] 1. Diongue K, Boye A, Bréchard L, et al.: Dermatophytic mycetoma of the scalp due to an atypical strain of Microsporum audouinii identified by MALDI-TOF MS and ITS sequencing. J. Mycol. Médicale. 2019 Jun; 29(2): 185–188. PubMed Abstract | Publisher Full Text

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Diagnosis and Molecular Characterization of Dermatophytosis: An Observational Study

Abstract

Keywords

Revised Amendments from Version 1

Introduction

Protocol

Study design

Study setting

Eligibility criteria

Data management

Sample size

Reagents required

Instruments required

Protocol

Statistical analysis plan

Study status

Ethical considerations

Discussion

Data availability

Acknowledgments

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