caa a22 4500 605498091 CHVBK 20210128100544.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00253-014-6246-x doi (NATIONALLICENCE)springer-10.1007/s00253-014-6246-x Fast and highly specific DNA-based multiplex detection on a solid support [Elektronische Daten] [Ivan Barišić, Verena Kamleithner, Silvia Schönthaler, Herbert Wiesinger-Mayr] Highly specific and fast multiplex detection methods are essential to conduct reasonable DNA-based diagnostics and are especially important to characterise infectious diseases. More than 1000 genetic targets such as antibiotic resistance genes, virulence factors and phylogenetic markers have to be identified as fast as possible to facilitate the correct treatment of a patient. In the present work, we developed a novel ligation-based DNA probe concept that was combined with the microarray technology and used it for the detection of bacterial pathogens. The novel linear chain (LNC) probes identified all tested species correctly within 1h based on their 16S rRNA gene in a 25-multiplex reaction. Genomic DNA was used directly as template in the ligation reaction identifying as little as 107 cells without any pre-amplification. The high specificity was further demonstrated characterising a single nucleotide polymorphism leading to no false positive fluorescence signals of the untargeted single nucleotide polymorphism (SNP) variants. In comparison to conventional microarray probes, the sensitivity of the novel LNC3 probes was higher by a factor of 10 or more. In summary, we present a fast, simple, highly specific and sensitive multiplex detection method adaptable for a wide range of applications. Springer-Verlag Berlin Heidelberg, 2014 Molecular biology nationallicence DNA microarray nationallicence Solid support-based ligation reaction nationallicence Multiplex detection nationallicence Pathogen characterisation nationallicence Barišić Ivan Molecular Diagnostics, AIT Austrian Institute of Technology, Muthgasse 11, 1190, Vienna, Austria aut Kamleithner Verena Molecular Diagnostics, AIT Austrian Institute of Technology, Muthgasse 11, 1190, Vienna, Austria aut Schönthaler Silvia Molecular Diagnostics, AIT Austrian Institute of Technology, Muthgasse 11, 1190, Vienna, Austria aut Wiesinger-Mayr Herbert Molecular Diagnostics, AIT Austrian Institute of Technology, Muthgasse 11, 1190, Vienna, Austria aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/1(2015-01-01), 413-423 0175-7598 99:1<413 2015 99 253 https://doi.org/10.1007/s00253-014-6246-x 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/s00253-014-6246-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Barišić Ivan Molecular Diagnostics, AIT Austrian Institute of Technology, Muthgasse 11, 1190, Vienna, Austria aut NATIONALLICENCE 700 1- Kamleithner Verena Molecular Diagnostics, AIT Austrian Institute of Technology, Muthgasse 11, 1190, Vienna, Austria aut NATIONALLICENCE 700 1- Schönthaler Silvia Molecular Diagnostics, AIT Austrian Institute of Technology, Muthgasse 11, 1190, Vienna, Austria aut NATIONALLICENCE 700 1- Wiesinger-Mayr Herbert Molecular Diagnostics, AIT Austrian Institute of Technology, Muthgasse 11, 1190, Vienna, Austria aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/1(2015-01-01), 413-423 0175-7598 99:1<413 2015 99 253