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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Barisic, Ivan Schoenthaler, Silvia Ke, Rongqin Nilsson, Mats Noehammer, Christa Wiesinger-Mayr, Herbert |
| Description | Author Affiliation: Barisic I ( AIT Austrian Institute of Technology, Molecular Diagnostics, 1190 Vienna, Austria. Electronic address: ivan.barisic@ait.ac.at.) |
| Abstract | The elucidation of resistance mechanisms is of central importance to providing and maintaining efficient medical treatment. However, molecular detection methods covering the complete set of resistance genes with a single test are still missing. Here, we present a novel 100-plex assay based on padlock probes in combination with a microarray that allows the simultaneous large-scale identification of highly diverse ß-lactamases. The specificity of the assay was performed using 70 clinical bacterial isolates, recovering 98% of the ß-lactamase nucleotide sequences present. Additionally, the sensitivity was evaluated with PCR products and genomic bacterial DNA, revealing a detection limit of 10(4) DNA copies per reaction when using PCR products as the template. Pre-amplification of genomic DNA in a 25-multiplex PCR further facilitated the detection of ß-lactamase genes in dilutions of 10(7) cells/mL. In summary, we present an efficient, highly specific, and highly sensitive multiplex detection method for any gene. |
| File Format | HTM / HTML |
| ISSN | 07328893 |
| Issue Number | 2 |
| Volume Number | 77 |
| e-ISSN | 18790070 |
| Journal | Diagnostic Microbiology and Infectious Disease |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2013-10-01 |
| Publisher Place | United States |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Microbiology Discipline Communicable Diseases Bacterial Proteins Genetics Dna Probes Drug Resistance, Bacterial Multiplex Polymerase Chain Reaction Methods Oligonucleotide Array Sequence Analysis Beta-lactamases Cluster Analysis Dna, Bacterial Analysis Enterobacteriaceae Enterobacteriaceae Infections Microbiology Humans Limit Of Detection Molecular Typing Journal Article Research Support, Non-u.s. Gov't |
| Content Type | Text |
| Resource Type | Article |
| Subject | Infectious Diseases Microbiology (medical) |
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