P2399 Lateral Flow Immunoassay for Rapid Detection of Dermatophytes in Clinical Specimens

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P2399 Lateral flow immunoassay for rapid detection of dermatophytes in clinical specimens Amanda Burnham-Marusich*1, Caitlyn Orne1 2, Alexander Kvam3, Heather Green2, Alexandra Myers4, Aline Rodrigues Hoffmann4, Amy Crum5, Mahmoud Ghannoun6, Thomas Kozel1 3 1DxDiscovery, Reno, United States, 2University of Nevada, Reno, Reno, United States, 3University of Nevada, Reno School of Medicine, Reno, United States, 4Texas A&M University, College Station, United States, 5Houston SPCA, Houston, United States, 6Case Western Reserve University, Cleveland, United States Background: Fungal skin infections affect approximately 1 billion people each year. Most infections are treated with topical antifungal agents. Other infections, e.g., tinea capitis and onychomycosis, require use of oral antifungal agents that may produce significant side effects in some patients. Laboratory confirmation of fungal infection prior to use of antifungal agents is recommended but often not done due, in part, to the time and cost of laboratory testing. The goal of this study was to develop a lateral flow immunoassay (LFIA) for rapid diagnosis of dermatophytosis. Materials/methods: Monoclonal antibody (mAb) 2DA6 was produced from splenocytes of mice immunized with fungal cell wall fragments. Hybridomas were generated using standard methods. Results: A LFIA was constructed from mAb 2DA6 for detection of mannans of dermatophyte fungi in clinical samples. The antibody is reactive with the alpha-1,6 mannose backbone in mannans of fungi of the Zygomycota and the Ascomycota. However, mAb 2DA6 has an exquisite sensitivity for mannans of dermatophytes and the Zygomycota due to an apparent low level of side chain substitution found on these mannans vs. high levels of side chain substitution that occludes the backbone in mannans of other fungi. The dermatophyte LFIA was first evaluated using extracts from laboratory cultures of seven different dermatophytes that produce infection in man and animals. All extracts produced positive results in 15 min or less. The LFIA was next evaluated with hair and skin scrapings from dogs and cats with skin lesions. Results were compared to culture. Results of testing of six culture-positive and six culture- negative animals showed a diagnostic sensitivity of 100% and a specificity of 83%. Finally, the LFIA was tested with nail scrapings from normal subjects or individuals whose scrapings were positive by culture and/or microscopy. LFIA evaluation of specimens from infected individuals produced positive results. Conclusions: An LFIA constructed from mAb 2DA6 rapidly detected dermatophyte mannan in extracts from lab-grown cultures and clinical specimens. The availability of a rapid test for diagnosis of dermatophyte infection may increase compliance with recommendations for laboratory confirmation of infection prior to initiation of oral antifungal therapy for tinea capitis and onychomycosis. .

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