CARPEGEN Myco Diagnostics DNA Chips for Diagnosing Fungal Infections Max Koltzscher & Antje Rötger Carpegen GmbH, Münster
Carpegen: Company Profile molecular diagnostics using Real-time PCR and DNA chip technology focus on infections in dentistry & gynecology R&D financed by national grants (BMBF) European dental market: close cooperation with GABA International certified quality management system (DIN EN ISO 9001:2000) www.carpegen.com
Worldwide Increase in Superficial and Invasive Mycoses rising numbers of immune-compromised patients more elderly patients increased use of antibiotics tropical tourism Species identification is essential for: choice of an efficient therapy predicting/monitoring course of disease epidemiological reasons
Conventional Diagnosis of Fungal Infections Diagnosis is mainly based on morphological and physiological features. up to several weeks until diagnosis ambiguous results are possible strongly dependent on examiner's expertise: standardization practically impossible Rapid, unambiguous and objective tests are needed.
DNA Chip Technology as the New Method of Choice simultaneous detection of many pathogens very sensitive when combined with upstream PCR rapid diagnostics (< 1 day) perfect for numerous features: cost-efficient detection of mixed infections uncomplicated expansion highly objective, standardized diagnosis
Detection of Fungal Pathogens using Chip Technology: PCR and Labeling Primer 1 rdna 18S rdna 5,8S 28S rdna Primer 2 PCR using universal primers against conserved rdna regions PCR product comprises highly variable und regions labeling with a fluorescent dye
Detection of Fungal Pathogens using Chip Technology: Design and Spotting of Probes Seq 1 Seq 2 Seq 3 Seq 4 CAATCTCGGACTGCTGCATCTCGCTGTGTAGAC-TG--GA-GAAGTGCGCTTAAGCATATCAATAAGC CAATCTCGGACTGCTGCATCTCGCTGTGTTGAC---TTGA-GAAGTGCGCTTAAGCATATCAATAAGC CAATCTCGGACTGCTGCATCTCACTGGTCCA--GTGTTGAAGTTCGT-GCTTAAGCATATCAATAAGC CAATCTCGGACTGCTGCATCTCACTGGTCCATGGTGTTGAAGCGT---GCTTAAGCATATCAATAAGC rdna 18S rdna 5,8S 28S rdna DNA probes DNA probes design of specific DNA probes against highly variable regions multiple probes per species for high specificity spotting of probes on modified glass slides (= microarrays)
Detection of Fungal Pathogens using Chip Technology: Hybridization and Scanning fluorescently labeled PCR products analysis of signal intensities hybridization laser scanning
Specifications easily distinguish between very closely related species threshold for detection is 100 fungal cells complete analysis takes approx. 12 hours (hands-on time is 3-4 hours) Superior to conventional methods in terms of: specificity sensitivity speed objectivity
Discrimination between Closely Related Species Candida albicans Candida dubliniensis Candida africana shared probes
Current Commercialization as a Service CARPEGEN Myco Diagnostics Oral for diagnosing oral mycoses CARPEGEN Myco Diagnostics Gyn for diagnosing genital mycoses both detect 12 pathogenic yeasts CARPEGEN Myco Diagnostics sampling set Candida albicans Candida lusitaniae Candida glabrata Candida guilliermondii Candida tropicalis Candida dubliniensis Candida krusei Candida kefyr Candida parapsilosis Candida africana Candida famata Geotrichum candidum Current Customers: dentists gynecologists
Clinical Applications Dentistry/Oral Diseases Gynecology diagnosis of different candidiases prior to antibiotic therapy mucosal lesions of unclear origin recurring/chronic fungal infections for choice and monitoring success of antifungal therapy in case of failing local therapy for differential diagnosis: fungal versus bacterial/viral infection prevention of Candida colonization/infection of newborns recurring/chronic infections for choice and monitoring success of antifungal therapy + future applications in intensive care settings and dermatology
Partnering Interests 1. partners for co-developing diagnostic kits containing our DNA chips 2. partners for R&D projects concerning new potential areas of use (e.g. veterinary medicine, environmental diagnostics, etc.) 3. marketing/distribution partners for current and upcoming DNA chips