Internal Transcribed Spacer RDNA And TEF-1α Gene Sequencing Of Pathogenic Dermatophyte Species And Differentiation Of Closely Related Species Using PCR-RFLP Of The Topoisomerase II
Objective Precise identification of dermatophyte species significantly improves treatment and controls measures of dermatophytosis in human and animals. This study was designed to evaluate molecular tools effectiveness of the gene sequencing and DNA-based fragment polymorphism analysis for accurate...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Royan Institute (ACECR), Tehran
2020-04-01
|
| Series: | Cell Journal |
| Subjects: | |
| Online Access: | https://celljournal.org/journal/article/fulltext/pcr-rflp-targeting-of-the-dna-topoisomerase-ii-for-the-identification-of-the-common-dermatophyte-species.pdf |
| _version_ | 1819087359351717888 |
|---|---|
| author | Zahra Salehi Masoomeh Shams-Ghahfarokhi, Mehdi Razzaghi-Abyaneh |
| author_facet | Zahra Salehi Masoomeh Shams-Ghahfarokhi, Mehdi Razzaghi-Abyaneh |
| author_sort | Zahra Salehi |
| collection | DOAJ |
| description | Objective
Precise identification of dermatophyte species significantly improves treatment and controls measures of dermatophytosis in human and animals. This study was designed to evaluate molecular tools effectiveness of the gene sequencing and DNA-based fragment polymorphism analysis for accurate identification and differentiation of closely- related dermatophyte species isolated from clinical cases of dermatophytosis and their antifungal susceptibility to the current antifungal agents.
Materials And Methods
In this experimental study, a total of 95 skin samples were inoculated into mycobiotic agar for two weeks at 28˚C. Morphological characteristics of the isolated dermatophytes were evaluated. DNA was extracted from the fungal culture for amplification of topoisomerase II gene fragments and polymerase chain reaction (PCR) products were digested by Hinf I enzyme. Internal transcribed spacer (ITS) rDNA and TEF-1α regions of the all isolates were amplified using the primers of ITS1/4 and EF-DermF/EF-DermR, respectively.
Results
Based on the morphological criteria, 24, 24, 24 and 23 isolates were identified as T. rubrum, T. interdigitale, T. tonsurans and E. floccosum, respectively. PCR-restriction fragment length polymorphism (RFLP) results provided identification pattern of the isolates for T. rubrum (19 isolates), T. tonsurans (28 isolates), T. interdigitale (26 isolates) and E. floccosum (22 isolates). Concatenated dataset results were similar in PCR-RFLP, except six T. interdigitale isolates belonging to T. mentagrophytes.
Conclusion
Our results clearly indicated that conventional morphology and PCR-RFLP were not able to precisely identify all dermatophyte species and differentiation of closely related species like T. interdigitale and T. mentagrophytes, while ITS rDNA and TEF-1α gene sequence analyses provided accurate identification of all isolates at the genus and species level. |
| first_indexed | 2024-12-21T21:34:53Z |
| format | Article |
| id | doaj.art-fb5a74baec954871bdd2fa687290509c |
| institution | Directory Open Access Journal |
| issn | 2228-5806 2228-5814 |
| language | English |
| last_indexed | 2024-12-21T21:34:53Z |
| publishDate | 2020-04-01 |
| publisher | Royan Institute (ACECR), Tehran |
| record_format | Article |
| series | Cell Journal |
| spelling | doaj.art-fb5a74baec954871bdd2fa687290509c2022-12-21T18:49:32ZengRoyan Institute (ACECR), TehranCell Journal2228-58062228-58142020-04-01221859110.22074/cellj.2020.6372Internal Transcribed Spacer RDNA And TEF-1α Gene Sequencing Of Pathogenic Dermatophyte Species And Differentiation Of Closely Related Species Using PCR-RFLP Of The Topoisomerase IIZahra Salehi0Masoomeh Shams-Ghahfarokhi,1Mehdi Razzaghi-Abyaneh2Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranDepartment of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IranDepartment of Mycology, Pasteur Institute of Iran, Tehran, IranObjective Precise identification of dermatophyte species significantly improves treatment and controls measures of dermatophytosis in human and animals. This study was designed to evaluate molecular tools effectiveness of the gene sequencing and DNA-based fragment polymorphism analysis for accurate identification and differentiation of closely- related dermatophyte species isolated from clinical cases of dermatophytosis and their antifungal susceptibility to the current antifungal agents. Materials And Methods In this experimental study, a total of 95 skin samples were inoculated into mycobiotic agar for two weeks at 28˚C. Morphological characteristics of the isolated dermatophytes were evaluated. DNA was extracted from the fungal culture for amplification of topoisomerase II gene fragments and polymerase chain reaction (PCR) products were digested by Hinf I enzyme. Internal transcribed spacer (ITS) rDNA and TEF-1α regions of the all isolates were amplified using the primers of ITS1/4 and EF-DermF/EF-DermR, respectively. Results Based on the morphological criteria, 24, 24, 24 and 23 isolates were identified as T. rubrum, T. interdigitale, T. tonsurans and E. floccosum, respectively. PCR-restriction fragment length polymorphism (RFLP) results provided identification pattern of the isolates for T. rubrum (19 isolates), T. tonsurans (28 isolates), T. interdigitale (26 isolates) and E. floccosum (22 isolates). Concatenated dataset results were similar in PCR-RFLP, except six T. interdigitale isolates belonging to T. mentagrophytes. Conclusion Our results clearly indicated that conventional morphology and PCR-RFLP were not able to precisely identify all dermatophyte species and differentiation of closely related species like T. interdigitale and T. mentagrophytes, while ITS rDNA and TEF-1α gene sequence analyses provided accurate identification of all isolates at the genus and species level.https://celljournal.org/journal/article/fulltext/pcr-rflp-targeting-of-the-dna-topoisomerase-ii-for-the-identification-of-the-common-dermatophyte-species.pdfDermatophytesGene SequencingPolymerase Chain Reaction-Restriction Fragment Length PolymorphismTopoisomerase II |
| spellingShingle | Zahra Salehi Masoomeh Shams-Ghahfarokhi, Mehdi Razzaghi-Abyaneh Internal Transcribed Spacer RDNA And TEF-1α Gene Sequencing Of Pathogenic Dermatophyte Species And Differentiation Of Closely Related Species Using PCR-RFLP Of The Topoisomerase II Cell Journal Dermatophytes Gene Sequencing Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Topoisomerase II |
| title | Internal Transcribed Spacer RDNA And TEF-1α Gene Sequencing Of Pathogenic Dermatophyte Species And Differentiation Of Closely Related Species Using PCR-RFLP Of The Topoisomerase II |
| title_full | Internal Transcribed Spacer RDNA And TEF-1α Gene Sequencing Of Pathogenic Dermatophyte Species And Differentiation Of Closely Related Species Using PCR-RFLP Of The Topoisomerase II |
| title_fullStr | Internal Transcribed Spacer RDNA And TEF-1α Gene Sequencing Of Pathogenic Dermatophyte Species And Differentiation Of Closely Related Species Using PCR-RFLP Of The Topoisomerase II |
| title_full_unstemmed | Internal Transcribed Spacer RDNA And TEF-1α Gene Sequencing Of Pathogenic Dermatophyte Species And Differentiation Of Closely Related Species Using PCR-RFLP Of The Topoisomerase II |
| title_short | Internal Transcribed Spacer RDNA And TEF-1α Gene Sequencing Of Pathogenic Dermatophyte Species And Differentiation Of Closely Related Species Using PCR-RFLP Of The Topoisomerase II |
| title_sort | internal transcribed spacer rdna and tef 1α gene sequencing of pathogenic dermatophyte species and differentiation of closely related species using pcr rflp of the topoisomerase ii |
| topic | Dermatophytes Gene Sequencing Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Topoisomerase II |
| url | https://celljournal.org/journal/article/fulltext/pcr-rflp-targeting-of-the-dna-topoisomerase-ii-for-the-identification-of-the-common-dermatophyte-species.pdf |
| work_keys_str_mv | AT zahrasalehi internaltranscribedspacerrdnaandtef1agenesequencingofpathogenicdermatophytespeciesanddifferentiationofcloselyrelatedspeciesusingpcrrflpofthetopoisomeraseii AT masoomehshamsghahfarokhi internaltranscribedspacerrdnaandtef1agenesequencingofpathogenicdermatophytespeciesanddifferentiationofcloselyrelatedspeciesusingpcrrflpofthetopoisomeraseii AT mehdirazzaghiabyaneh internaltranscribedspacerrdnaandtef1agenesequencingofpathogenicdermatophytespeciesanddifferentiationofcloselyrelatedspeciesusingpcrrflpofthetopoisomeraseii |