A novel universal DNA labeling and amplification system for rapid microarray-based detection of 117 antibiotic resistance genes in Gram-positive bacteria.

A rapid and simple DNA labeling system has been developed for disposable microarrays and has been validated for the detection of 117 antibiotic resistance genes abundant in Gram-positive bacteria. The DNA was fragmented and amplified using phi-29 polymerase and random primers with linkers. Labeling and further amplification were then performed by classic PCR amplification using biotinylated primers specific for the linkers. The microarray developed by Perreten et al. (Perreten, V., Vorlet-Fawer, L., Slickers, P., Ehricht, R., Kuhnert, P., Frey, J., 2005. Microarray-based detection of 90 antibiotic resistance genes of gram-positive bacteria. J.Clin.Microbiol. 43, 2291-2302.) was improved by additional oligonucleotides. A total of 244 oligonucleotides (26 to 37 nucleotide length and with similar melting temperatures) were spotted on the microarray, including genes conferring resistance to clinically important antibiotic classes like β-lactams, macrolides, aminoglycosides, glycopeptides and tetracyclines. Each antibiotic resistance gene is represented by at least 2 oligonucleotides designed from consensus sequences of gene families. The specificity of the oligonucleotides and the quality of the amplification and labeling were verified by analysis of a collection of 65 strains belonging to 24 species. Association between genotype and phenotype was verified for 6 antibiotics using 77 Staphylococcus strains belonging to different species and revealed 95% test specificity and a 93% predictive value of a positive test. The DNA labeling and amplification is independent of the species and of the target genes and could be used for different types of microarrays. This system has also the advantage to detect several genes within one bacterium at once, like in Staphylococcus aureus strain BM3318, in which up to 15 genes were detected. This new microarray-based detection system offers a large potential for applications in clinical diagnostic, basic research, food safety and surveillance programs for antimicrobial resistance.

Ms. hebr. oct. 117 - Ṿikuaḥ ha-torani ʹim ha-filosof

Vorbesitzer: Eljāqīm Carmoly; Abraham Merzbacher

Ms. lat. qu. 117 - Venezianische Geschichte. Kirchengeschichte. Türken. Ratschläge für Diplomaten

Vorbesitzer: Johann Hieronymus Zum Jungen

Ms. Barth. 117 (Ausst. 21) - Epistolae Pauli Apostoli cum glossa

Vorbesitzer: Bartholomaeusstift Frankfurt am Main

Na 50 Nachlass Arthur Schopenhauer - 'Schopenhauer-Archiv', 117 - Bitte um Zusendung eines Buches

veröffentlicht in: Schopenhauer, Arthur : Arthur Schopenhauers sämtliche Werke - München : Piper - Bd. 14 : Der Briefwechsel Arthur Schopenhauers ; 1 (1799 - 1849), Nr. 56;

Ms. Praed. 117 - Opus tabulare super quattuor specula Vincentii Bellovacensis

Vorbesitzer: Dominikanerkloster Frankfurt am Main

Na 50 Nachlass Arthur Schopenhauer - 'Schopenhauer-Archiv', 728 - Antwortschreiben zur Frage nach dem geeigneten juristischen Beistand in den laufenden Erbschaftsangelegenheiten

u.a.: Hohenfeldtsche Erben; Juristische Beratung durch Martin Emden; negative Äußerungen von Arthur Schopenhauer gegenüber der Frankfurt Gesellschaft; Halleysche Komet;

Ms.Ff.F.Stoltze 3. 117 - Brief von K. Hensel an Friedrich Stoltze

Bitte um Texte oder bibliographische Hinweise zu Stoltzes Veröffentlichungen über Goethe, für eine "Goethe-Arbeit", Bitte um biographische Notizen zu Marquard Georg Seufferhe1d

Ausst. 117/118 - Hl. Georg im Kampfe mit dem Drachen. Hl. Wolfgang : Teigdrucke

Vorbesitzer: Dominikanerkloster Frankfurt am Main;

Ms. lat. oct. 117 - Quadragesimale

Vorbesitzer: Gesellschaft der Freunde der Frankfurter Stadtbibliothek (Stifter)