Multiplex characterization of human pathogens including species and antibiotic-resistance gene identification
Barišić, Ivan and Petzka, Josefine and Schoenthaler, Silvia and Vierlinger, Klemens and Noehammer, Christa and Wiesinger-Mayr, Herbert,, 65, 48-55 (2016),
doi = https://doi.org/10.1099/jmm.0.000192,
publicationName = Microbiology Society,
issn = 0022-2615,
abstract= The efficient medical treatment of infections requires detailed information about the pathogens involved and potential antibiotic-resistance mechanisms. The dramatically increasing incidence of multidrug-resistant bacteria especially highlights the importance of sophisticated diagnostic tests enabling a fast patient-customized therapy. However, the current molecular detection methods are limited to either the detection of species or only a few antibiotic-resistance genes. In this work, we present a human pathogen characterization assay using a rRNA gene microarray identifying 75 species comprising bacteria and fungi. A statistical classifier was developed to facilitate the automated species identification. Additionally, the clinically most important β-lactamases were identified simultaneously in a 100-plex reaction using padlock probes and the same microarray. The specificity and sensitivity of the combined assay was determined using clinical isolates. The detection limit was 105 c.f.u. ml− 1, recovering 89 % of the detectable β-lactamase-encoding genes specifically. The total assay time was less than 7 h and the modular character of the antibiotic-resistance detection allows the easy integration of further genetic targets. In summary, we present a fast, highly specific and sensitive multiplex pathogen characterization assay.,
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