interdigitale grew in culture and also identified a dermatophyte species in an additional 32 specimens that were negative in Adriamycin molecular weight microscopy and culture. None of the investigated non-dermatophytic strains was positive. Sensitivity of MX PCR was higher as compared to mycological examination (97% vs. 81.1%). MX PCR for direct detection of dermatophytes from nail samples yielded mixed flora in 32.8% of samples. MX PCR proved sensitive and adequate for the diagnosis of dermatophytic onychomycosis. It is much adapted to cases where culture is negative or contaminated by overgrowing moulds, which makes

the identification of the causal agent problematic. Onychomycosis is the most common nail disease. It Crizotinib cost is mainly caused by dermatophytes of the genus Trichophyton. However, various non-dermatophyte filamentous fungi are often isolated from nails and usually considered as transient contaminants and are not the actual aetiological agent. Treatment of onychomycosis is closely linked to the identity of the causative agent, particularly in terms of whether or not it is a dermatophyte.[1] Onychomycosis is routinely diagnosed by mycological examination, which includes direct examination and culture.[2] Direct microscopic examination of infected nail material may give false positive results as it is usually enable to differentiate between dermatophyte and non-dermatophyte filamentous

fungi. Conventionally the definitive diagnosis is based on culture isolation, but culture of toenails is often associated with contaminants. Furthermore, morphological and physiological http://www.selleck.co.jp/products/Verteporfin(Visudyne).html characteristics may vary; the phenotypic features can be influenced by factors such as temperature variation and medium.[3] Routine identification by microscopy and culture requires considerable training of personnel and considerable supervisory expertise. Molecular approaches have been developed to provide more rapid and accurate alternatives for dermatophyte

identification. These methods include restriction fragment length polymorphism analysis RFLP,[1, 4, 5] PCR-ELISA,[6] double-round PCR,[7] nested PCR,[8, 9] real-time PCR,[10, 11] PCR using (GACA)4 primer,[12] sequencing.[13, 14] The main targets have been the following genes or DNA fragments: the ribosomal DNA region,[1, 3, 10, 13, 15, 16] DNA topoisomerase II genes,[6] actin gene[7] and the chitin synthase gene.[8, 14, 17, 18] In recent years, the multiplex (MX) PCR assay has been used to identify a great variety of fungi including dermatophytes.[15-21] However, Trichophyton mentagrophytes complex was not included in these studies despite its high frequency in nail infection.[14, 22, 23] The primary aim of this study was to design and develop a MX PCR assay to identify dermatophytes on one hand and T. rubrum and T. mentagrophytes complex on the other hand directly from nails samples.