Identification and differentiation of Trichophyton rubrum clinical isolates using PCR-RFLP and RAPD methods.

Trichophyton rubrum represents the most frequently isolated causative agent of superficial dermatophyte infections. Several genotyping methods have recently been introduced to improve the delineation between pathogenic fungi at both the species and the strain levels. The purpose of this study was to apply selected DNA fingerprinting methods to the identification and strain discrimination of T. rubrum clinical isolates. Fifty-seven isolates from as many tinea patients were subjected to species identification by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis and strain differentiation using a randomly amplified polymorphic DNA (RAPD) method, with two primers designated 1 and 6. Using PCR-RFLP, 55 of the isolates studied were confirmed to be T. rubrum. Among those, a total of 40 and five distinct profiles were obtained by RAPD with primers 1 and 6, respectively. The combination of profiles from both RAPD assays resulted in 47 genotypes and an overall genotypic diversity rate of 85.4%. A dendrogram analysis performed on the profiles generated by RAPD with primer 1 showed most of the isolates (87.3%) to be genetically related. PCR-RFLP serves as a rapid and reliable method for the identification of T. rubrum species, while the RAPD analysis is rather a disadvantageous tool for T. rubrum strain typing.

Treatment of cutaneous T-cell lymphomas with purine analogues (fludarabine and 2-chlorodeoxyadenosine).

PURPOSE: The aim of this study was to evaluate the efficacy and toxicity of the purine analogues 2-chlorodeoxyadenosine (2-CdA) and fludarabine (FAMP) combined with cyclophosphamide (CY) in the treatment of stage III and IV cutaneous T-cell lymphoma (CTCL). PATIENTS AND METHODS: From January 1998 to December 2002, 10 heavily pretreated patients with CTCL, hospitalized at the Department of Dermatology in Wroclaw, were administered monotherapy with 2-CdA (6 patients) or FAMP plus CY combination chemotherapy (4 patients). 2-CdA was administered at a dose of 0.12 mg/kg daily for 7 days every 28 days. FAMP was administered at a dose of 25 mg/m(2) daily, days 1-3, and cyclophosphamide 400 mg/m(2), day 1. The combination was repeated every 28 days. RESULTS: Five out of 6 patients treated with 2-CdA showed a transient partial remission of the skin lesions lasting for a median of 2 months, and 1 patient showed disease progression with dissemination of the skin lesions. Of the 4 patients who received FAMP plus CY 1 achieved complete remission lasting for 6 months, and 2 attained a partial response lasting for a median of 3 months. CONCLUSION: Purine analogues such as 2-CdA and FAMP may be used in the treatment of advanced stages of CTCL. The combination of FAMP plus CY, based on the restrictive effect of FAMP on the repair mechanisms of DNA damaged by CY, seems to be a promising therapeutic modality. Decreased immunity, leucopenia, thrombocytopenia and anaemia are common side effects of 2-CdA and FAMP.