Specific Multiplex Analysis of Pathogens Using a Direct 165 rRNA Hybridization in Microarray System

Title
Specific Multiplex Analysis of Pathogens Using a Direct 165 rRNA Hybridization in Microarray System
Author(s)
서정현황병희[황병희]신화희[신화희]차형준[차형준]
Keywords
OLIGONUCLEOTIDE MICROARRAYS; GENE-EXPRESSION; DNA MICROARRAY; UNITED-STATES; PCR; COMMUNITIES; BIOSENSORS; SEQUENCE; PROBE
Issue Date
201206
Publisher
AMER CHEMICAL SOC
Citation
ANALYTICAL CHEMISTRY, v.84, no.11, pp.4873 - 4879
Abstract
For the rapid multiplex analysis of pathogens, 16S rRNAs from cell lysates were directly applied onto a DNA microarray at room temperature (RT) for RNA-DNA hybridization. To eliminate the labeling step, seven fluorescent-labeled detector probes were cohybridized with 16S rRNA targets and adjacent specific capture probes. We found that eight pathogens were successfully discriminated by the 16S rRNA-based direct method, which showed greater specificity than the polymerase chain reaction (PCR)-labeled method due to chaperone and distance effects. A new specificity criterion for a perfect match between RNA and DNA was suggested to be 21-41% dissimilarity using correlation analysis between the mismatch and the sequence according to the guanine cytosine (GC) percentage or the distribution of mismatches. Six categories of food matrix (egg, meat, milk, rice, vegetable, and mixed) were also tested, and the target pathogen was successfully discriminated within statistically significant levels. Finally, we found that the intrinsic abundance of 16S rRNA molecules successfully substituted PCR-based amplification with a low limit of detection of 10-10(3) cells mL(-1) and a high quantitative linear correlation. Collectively, our suggested 16S rRNA-based direct method enables the highly sensitive, specific, and quantitative analysis of selected pathogens at RT within 2 h, even in food samples.
URI
http://hdl.handle.net/YU.REPOSITORY/28013http://dx.doi.org/10.1021/ac300476k
ISSN
0003-2700
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공과대학 > 화학공학부 > Articles
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