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Species-identification of dermatophytes Trichophyton, Microsporum and Epidermophyton by PCR and PCR-RFLP targeting of the DNA topoisomerase II genes.
J Dermatol Sci. 2003 Oct; 33(1):41-54.JD

Abstract

BACKGROUND

We have focused on the DNA topoisomerase II genes of pathogenic fungi and have previously applied polymerase chain reaction (PCR)-based identification of several species including the some of the major dermatophyte species.

OBJECTIVE

To identify the dermatophytes (18 species) to a species level by PCR and PCR-restriction fragment length polymorphism (RFLP) techniques, without determining the nucleotide sequence.

METHODS

The genomic DNAs of the dermatophytes (ten species of Trichophyton, seven species of Microsporum, and Epidermaphyton floccosum) were amplified by PCR using a common primer set (dPsD1) for the dermatophytes, followed by nested PCR using other primer sets (dPsD2, PsT and PsME) that contained primers specific for the DNA topoisomerase II genes of the dermatophytes. PCRs using PsT and PsME were used for the species-identification of Trichophyton, Microsporum and E. floccosum. The PCR products generated by dPsD2 were digested with restriction enzymes (Hinc II, Hinf, Afl II and PflM I), and the restriction profiles were analyzed.

RESULTS

Of the eighteen species of dermatophytes, five species (T. rubrum, T. violaceum, M. canis, M. gypseum and E. floccosum) were specifically identified by the PCR using PsT and PsME to the species level, and the remaining species were identified by the unique restriction profiles for each species in the PCR-RFLP analysis, except that the restriction profile of T. mentagrophytes var. interdigitale was identical to that of T. mentagrophytes var. quinckeanum.

CONCLUSION

PCR and PCR-RFLP techniques targeting the DNA topoisomerase II gene are simple and rapid, and quite useful as tools for the identification of dermatophytes to the species level.

Authors+Show Affiliations

Division of Molecular Mycology and Medicine, Department of Advanced Medical Science, Center for Neural Disease and Cancer, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-8550, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

14527738

Citation

Kanbe, Toshio, et al. "Species-identification of Dermatophytes Trichophyton, Microsporum and Epidermophyton By PCR and PCR-RFLP Targeting of the DNA Topoisomerase II Genes." Journal of Dermatological Science, vol. 33, no. 1, 2003, pp. 41-54.
Kanbe T, Suzuki Y, Kamiya A, et al. Species-identification of dermatophytes Trichophyton, Microsporum and Epidermophyton by PCR and PCR-RFLP targeting of the DNA topoisomerase II genes. J Dermatol Sci. 2003;33(1):41-54.
Kanbe, T., Suzuki, Y., Kamiya, A., Mochizuki, T., Kawasaki, M., Fujihiro, M., & Kikuchi, A. (2003). Species-identification of dermatophytes Trichophyton, Microsporum and Epidermophyton by PCR and PCR-RFLP targeting of the DNA topoisomerase II genes. Journal of Dermatological Science, 33(1), 41-54.
Kanbe T, et al. Species-identification of Dermatophytes Trichophyton, Microsporum and Epidermophyton By PCR and PCR-RFLP Targeting of the DNA Topoisomerase II Genes. J Dermatol Sci. 2003;33(1):41-54. PubMed PMID: 14527738.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Species-identification of dermatophytes Trichophyton, Microsporum and Epidermophyton by PCR and PCR-RFLP targeting of the DNA topoisomerase II genes. AU - Kanbe,Toshio, AU - Suzuki,Yasuhiro, AU - Kamiya,Atsushi, AU - Mochizuki,Takashi, AU - Kawasaki,Masako, AU - Fujihiro,Machiko, AU - Kikuchi,Akihiko, PY - 2003/10/7/pubmed PY - 2004/5/27/medline PY - 2003/10/7/entrez SP - 41 EP - 54 JF - Journal of dermatological science JO - J. Dermatol. Sci. VL - 33 IS - 1 N2 - BACKGROUND: We have focused on the DNA topoisomerase II genes of pathogenic fungi and have previously applied polymerase chain reaction (PCR)-based identification of several species including the some of the major dermatophyte species. OBJECTIVE: To identify the dermatophytes (18 species) to a species level by PCR and PCR-restriction fragment length polymorphism (RFLP) techniques, without determining the nucleotide sequence. METHODS: The genomic DNAs of the dermatophytes (ten species of Trichophyton, seven species of Microsporum, and Epidermaphyton floccosum) were amplified by PCR using a common primer set (dPsD1) for the dermatophytes, followed by nested PCR using other primer sets (dPsD2, PsT and PsME) that contained primers specific for the DNA topoisomerase II genes of the dermatophytes. PCRs using PsT and PsME were used for the species-identification of Trichophyton, Microsporum and E. floccosum. The PCR products generated by dPsD2 were digested with restriction enzymes (Hinc II, Hinf, Afl II and PflM I), and the restriction profiles were analyzed. RESULTS: Of the eighteen species of dermatophytes, five species (T. rubrum, T. violaceum, M. canis, M. gypseum and E. floccosum) were specifically identified by the PCR using PsT and PsME to the species level, and the remaining species were identified by the unique restriction profiles for each species in the PCR-RFLP analysis, except that the restriction profile of T. mentagrophytes var. interdigitale was identical to that of T. mentagrophytes var. quinckeanum. CONCLUSION: PCR and PCR-RFLP techniques targeting the DNA topoisomerase II gene are simple and rapid, and quite useful as tools for the identification of dermatophytes to the species level. SN - 0923-1811 UR - https://www.unboundmedicine.com/medline/citation/14527738/Species_identification_of_dermatophytes_Trichophyton_Microsporum_and_Epidermophyton_by_PCR_and_PCR_RFLP_targeting_of_the_DNA_topoisomerase_II_genes_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0923181103001506 DB - PRIME DP - Unbound Medicine ER -