Journal: PLoS ONE
Article Title: Fast DNA Serotyping and Antimicrobial Resistance Gene Determination of Salmonella enterica with an Oligonucleotide Microarray-Based Assay
doi: 10.1371/journal.pone.0046489
Figure Lengend Snippet: (a) Linear Multiplex Amplification starting from clonal RNA free genomic DNA, extracted DNA is internally labeled with biotin (Label [L]) and amplified in a linear multiplex PCR reaction; (b) Hybridization: the biotin labeled, single-stranded DNA product hybridizes specifically under stringent conditions to the corresponding probes. The resulting duplex is detected using a horse-radish peroxidase (Enzyme [E]) – streptavidin conjugate, which converts the substrate (Seramun green [S]) into a colored local precipitate. (c) Detection: the ArrayMate™ Reader (or ArrayTube™ Reader ATR 03) enables the visualization and subsequent automated analysis of the array image. The presence of a dark precipitated spot indicates successful hybridization; (d) Analysis: the assay specific software analysis script, supplied with the ArrayMate™ Reader (or ArrayTube™ Reader ATR 03), measures the signal intensity of each probe and determines with an assay specific algorithm which genes/alleles are present in the sample. (e) Genotype analysis: the PatternMatching software supplied with the ArrayMate™ Reader (or ArrayTube™ Reader ATR 03) is comparing the resulting pattern with a local database including 132 reference serovars previously sero- and genotyped, finally a report is given to which serovar the sample strain belongs with regard to the Kauffman-White Scheme.
Article Snippet: The visualization was achieved by adding 100 µl of peroxidase substrate D1 to the ArrayStrips, and signals were detected with the ArrayMate device (Alere Technologies, Jena, Germany) ( ).
Techniques: Multiplex Assay, Amplification, Labeling, Hybridization, Software
![(a) Linear Multiplex Amplification starting from clonal RNA free genomic DNA, extracted DNA is internally labeled with biotin (Label [L]) and amplified in a linear multiplex PCR reaction; (b) Hybridization: the biotin labeled, single-stranded DNA product hybridizes specifically under stringent conditions to the corresponding probes. The resulting duplex is detected using a horse-radish peroxidase (Enzyme [E]) – streptavidin conjugate, which converts the substrate (Seramun green [S]) into a colored local precipitate. (c) Detection: the <t>ArrayMate™</t> Reader (or ArrayTube™ Reader ATR 03) enables the visualization and subsequent automated analysis of the array image. The presence of a dark precipitated spot indicates successful hybridization; (d) Analysis: the assay specific software analysis script, supplied with the ArrayMate™ Reader (or ArrayTube™ Reader ATR 03), measures the signal intensity of each probe and determines with an assay specific algorithm which genes/alleles are present in the sample. (e) Genotype analysis: the PatternMatching software supplied with the ArrayMate™ Reader (or ArrayTube™ Reader ATR 03) is comparing the resulting pattern with a local database including 132 reference serovars previously sero- and genotyped, finally a report is given to which serovar the sample strain belongs with regard to the Kauffman-White Scheme.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_4306/pmc03464306/pmc03464306__pone.0046489.g001.jpg)