caa a22 4500 605501459 CHVBK 20210128100601.0 cr unu---uuuuu 210128e20150901xx s 000 0 eng 10.1007/s00253-015-6774-z doi (NATIONALLICENCE)springer-10.1007/s00253-015-6774-z Thirty-minute screening of antibiotic resistance genes in bacterial isolates with minimal sample preparation in static self-dispensing 64 and 384 assay cards [Elektronische Daten] [Tanja Kostić, Michael Ellis, Maggie Williams, Tiffany Stedtfeld, John Kaneene, Robert Stedtfeld, Syed Hashsham] In a clinical setting, molecular assays such as polymerase chain reaction offer a rapid means to infer or confirm identity and therapeutic decisions. Accordingly, a number of molecular assays targeting identity and antibiotic resistance (AR) genes have been developed; however, these methods can be technically complex and relatively expensive. Herein, we describe a diagnostic concept utilizing isothermal amplification technology with non-purified heat-lysed cells and self-dispensing cards for testing multiple primers in parallel. This proof-of-concept study, performed with Staphylococcus aureus isolates and associated AR genes, was compared with culture-based susceptibility and quantitative PCR (qPCR). Results demonstrate reduced sample processing steps resulting in a turnaround time (starting from bacterial culture to ending in the antibiotic resistance gene profile) in less than 30min. For antibiotics tested in which an associated AR gene was targeted on the Gene-Z card, 69% (18/26) of culture-based resistance events were positive for related AR genes. A comparison of loop-mediated isothermal amplification (LAMP) and qPCR assays targeting the same antibiotic resistance genes showed a 98.2% agreement in terms of presence and absence calls. Identity-based discrepancies between conventional (phenotypic) and molecular (genotypic) results were further resolved, and we were able to demonstrate higher accuracy in identification with the molecular analysis. The Author(s), 2015 Antibiotic resistance nationallicence Rapid genetic testing nationallicence LAMP nationallicence Point of care nationallicence Gene-Z nationallicence S. aureus nationallicence E. faecalis nationallicence E. faecium nationallicence Kostić Tanja AIT Austrian Institute of Technology GmbH, Bioresources Unit, Konrad Lorenz Strasse 24, A-3430, Tulln an der Donau, Austria aut Ellis Michael Civil and Environmental Engineering, Michigan State University, 48824, East Lansing, MI, USA aut Williams Maggie Civil and Environmental Engineering, Michigan State University, 48824, East Lansing, MI, USA aut Stedtfeld Tiffany Civil and Environmental Engineering, Michigan State University, 48824, East Lansing, MI, USA aut Kaneene John Center for Comparative Epidemiology, College of Veterinary Medicine, Michigan State University, 48824, East Lansing, MI, USA aut Stedtfeld Robert Civil and Environmental Engineering, Michigan State University, 48824, East Lansing, MI, USA aut Hashsham Syed Civil and Environmental Engineering, Michigan State University, 48824, East Lansing, MI, USA aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/18(2015-09-01), 7711-7722 0175-7598 99:18<7711 2015 99 253 https://doi.org/10.1007/s00253-015-6774-z text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6774-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kostić Tanja AIT Austrian Institute of Technology GmbH, Bioresources Unit, Konrad Lorenz Strasse 24, A-3430, Tulln an der Donau, Austria aut NATIONALLICENCE 700 1- Ellis Michael Civil and Environmental Engineering, Michigan State University, 48824, East Lansing, MI, USA aut NATIONALLICENCE 700 1- Williams Maggie Civil and Environmental Engineering, Michigan State University, 48824, East Lansing, MI, USA aut NATIONALLICENCE 700 1- Stedtfeld Tiffany Civil and Environmental Engineering, Michigan State University, 48824, East Lansing, MI, USA aut NATIONALLICENCE 700 1- Kaneene John Center for Comparative Epidemiology, College of Veterinary Medicine, Michigan State University, 48824, East Lansing, MI, USA aut NATIONALLICENCE 700 1- Stedtfeld Robert Civil and Environmental Engineering, Michigan State University, 48824, East Lansing, MI, USA aut NATIONALLICENCE 700 1- Hashsham Syed Civil and Environmental Engineering, Michigan State University, 48824, East Lansing, MI, USA aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/18(2015-09-01), 7711-7722 0175-7598 99:18<7711 2015 99 253