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Rapid real-time diagnostic PCR for Trichophyton rubrum and Trichophyton mentagrophytes in patients with tinea unguium and tinea pedis using specific fluorescent probes.
J Dermatol Sci. 2013 Mar; 69(3):229-35.JD

Abstract

BACKGROUND

Trichophyton rubrum and Trichophyton mentagrophytes human-type (synonym, Trichophyton interdigitale (anthropophilic)) are major causative pathogens of tinea unguium. For suitable diagnosis and treatment, rapid and accurate identification of etiologic agents in clinical samples using reliable molecular based method is required.

OBJECTIVE

For identification of organisms causing tinea unguium, we developed a new real-time polymerase chain reaction (PCR) with a pan-fungal primer set and probe, as well as specific primer sets and probes for T. rubrum and T. mentagrophytes human-type.

METHODS

We designed two sets of primers from the internal transcribed spacer 1 (ITS1) region of fungal ribosomal DNA (rDNA) and three quadruple fluorescent probes, one for detection wide range pathogenic fungi and two for classification of T. rubrum and T. mentagrophytes by specific binding to different sites in the ITS1 region. We investigated the specificity of these primer sets and probes using fungal genomic DNA, and also examined 42 clinical specimens with our real-time PCR.

RESULTS

The primers and probes specifically detected T. rubrum, T. mentagrophytes, and a wide range of pathogenic fungi. The causative pathogens were identified in 42 nail and skin samples from 32 patients. The total time required for identification of fungal species in each clinical specimen was about 3h. The copy number of each fungal DNA in the clinical specimens was estimated from the intensity of fluorescence simultaneously.

CONCLUSION

This PCR system is one of the most rapid and sensitive methods available for diagnosing dermatophytosis, including tinea unguium and tinea pedis.

Authors+Show Affiliations

Teikyo University Institute of Medical Mycology and Genome Research Center, Tokyo, Japan.No affiliation info availableNo 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

23287391

Citation

Miyajima, Yoshiharu, et al. "Rapid Real-time Diagnostic PCR for Trichophyton Rubrum and Trichophyton Mentagrophytes in Patients With Tinea Unguium and Tinea Pedis Using Specific Fluorescent Probes." Journal of Dermatological Science, vol. 69, no. 3, 2013, pp. 229-35.
Miyajima Y, Satoh K, Uchida T, et al. Rapid real-time diagnostic PCR for Trichophyton rubrum and Trichophyton mentagrophytes in patients with tinea unguium and tinea pedis using specific fluorescent probes. J Dermatol Sci. 2013;69(3):229-35.
Miyajima, Y., Satoh, K., Uchida, T., Yamada, T., Abe, M., Watanabe, S., Makimura, M., & Makimura, K. (2013). Rapid real-time diagnostic PCR for Trichophyton rubrum and Trichophyton mentagrophytes in patients with tinea unguium and tinea pedis using specific fluorescent probes. Journal of Dermatological Science, 69(3), 229-35. https://doi.org/10.1016/j.jdermsci.2012.11.589
Miyajima Y, et al. Rapid Real-time Diagnostic PCR for Trichophyton Rubrum and Trichophyton Mentagrophytes in Patients With Tinea Unguium and Tinea Pedis Using Specific Fluorescent Probes. J Dermatol Sci. 2013;69(3):229-35. PubMed PMID: 23287391.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Rapid real-time diagnostic PCR for Trichophyton rubrum and Trichophyton mentagrophytes in patients with tinea unguium and tinea pedis using specific fluorescent probes. AU - Miyajima,Yoshiharu, AU - Satoh,Kazuo, AU - Uchida,Takao, AU - Yamada,Tsuyoshi, AU - Abe,Michiko, AU - Watanabe,Shin-ichi, AU - Makimura,Miho, AU - Makimura,Koichi, Y1 - 2012/12/05/ PY - 2011/07/28/received PY - 2012/11/24/revised PY - 2012/11/27/accepted PY - 2013/1/5/entrez PY - 2013/1/5/pubmed PY - 2013/9/24/medline SP - 229 EP - 35 JF - Journal of dermatological science JO - J. Dermatol. Sci. VL - 69 IS - 3 N2 - BACKGROUND: Trichophyton rubrum and Trichophyton mentagrophytes human-type (synonym, Trichophyton interdigitale (anthropophilic)) are major causative pathogens of tinea unguium. For suitable diagnosis and treatment, rapid and accurate identification of etiologic agents in clinical samples using reliable molecular based method is required. OBJECTIVE: For identification of organisms causing tinea unguium, we developed a new real-time polymerase chain reaction (PCR) with a pan-fungal primer set and probe, as well as specific primer sets and probes for T. rubrum and T. mentagrophytes human-type. METHODS: We designed two sets of primers from the internal transcribed spacer 1 (ITS1) region of fungal ribosomal DNA (rDNA) and three quadruple fluorescent probes, one for detection wide range pathogenic fungi and two for classification of T. rubrum and T. mentagrophytes by specific binding to different sites in the ITS1 region. We investigated the specificity of these primer sets and probes using fungal genomic DNA, and also examined 42 clinical specimens with our real-time PCR. RESULTS: The primers and probes specifically detected T. rubrum, T. mentagrophytes, and a wide range of pathogenic fungi. The causative pathogens were identified in 42 nail and skin samples from 32 patients. The total time required for identification of fungal species in each clinical specimen was about 3h. The copy number of each fungal DNA in the clinical specimens was estimated from the intensity of fluorescence simultaneously. CONCLUSION: This PCR system is one of the most rapid and sensitive methods available for diagnosing dermatophytosis, including tinea unguium and tinea pedis. SN - 1873-569X UR - https://www.unboundmedicine.com/medline/citation/23287391/Rapid_real_time_diagnostic_PCR_for_Trichophyton_rubrum_and_Trichophyton_mentagrophytes_in_patients_with_tinea_unguium_and_tinea_pedis_using_specific_fluorescent_probes_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0923-1811(12)00934-6 DB - PRIME DP - Unbound Medicine ER -