Diagnostic Potential of Immunoblot Analysis for Identifying Rabbits Infected with Trichophyton mentagrophytes
Petra Zrimšek1* and Marinka Drobnič-Košorok2
1Clinic for Reproduction and Horses, Veterinary Faculty, University of Ljubljana, Gerbičeva 60, SI-1000 Ljubljana, Slovenia
2Institute of Physiology, Pharmacology and Toxicology, Veterinary Faculty, University of Ljubljana, Gerbičeva 60, SI-1000 Ljubljana, Slovenia
Article history:
Received: June 2, 2003
Accepted: November 10, 2003
Key words:
immunoblotting, diagnostic evaluation, rabbit, Trichophyton mentagrophytes
Summary:
The aim of this study was to investigate the diagnostic potential of immunoblot analysis for detecting rabbits naturally infected with Trichophyton mentagrophytes. A protein extract of the dermatophyte T. mentagrophytes was prepared from a fungal mat grown in liquid medium, where the production of keratinases was stimulated by keratin as a nitrogen source. The proteins derived from disrupted T. mentagrophytes were separated by SDS-PAGE and visualized by Western blotting. Twenty-one rabbits infected with T. mentagrophytes and 10 non-dermatophyte exposed rabbits were studied. Eight dominant bands were identified in immunoblots from dermatophyte-infected rabbits, with apparent molecular weights ranging from 20 to 75 kDa. The diagnostic potential of the bands for identifying infected rabbits was evaluated by determining sensitivity, specificity, positive and negative predictive values, and overall accuracy. Bands at 20, 30 and 33 kDa were observed with 100 % specificity and 76.19 % sensitivity and can thus be considered highly accurate in correct identification of the infected animals. Accuracies from 54 to 62 % were found for the other dominant bands. Bands at 42, 45 and 75 kDa with 85.71 to 100 % sensitivity appeared not to be specific for dermatophyte infection. Although the specificity of bands at 26 and 28 kDa was found to be 100 %, their accuracy was only moderate, due to the low level of sensitivity, 9.52 and 23.81 %, respectively.
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