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Title: SUSPENSION MICROARRAY WITH DENDRIMER SIGNAL AMPLIFICATION ALLOWS DIRECT AND HIGH THROUGHPUT SUBTYPING OF LISTERIA MONOCYTOGENES FROM GENOMIC DNA

Author
item Borucki, Monica
item Reynolds, James
item CALL, DOUGLAS - WSU
item Ward, Todd
item Page, Brent
item KADUSHIN, JAMES - GENISPHERE, INC.

Submitted to: Journal of Clinical Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/6/2005
Publication Date: 7/1/2005
Citation: Borucki, M.K., Reynolds, J.O., Call, D., Ward, T.J., Page, B.T., Kadushin, J. 2005. Suspension Microarray with Dendrimer Signal Amplification Allows Direct and High-Throughput Subtyping of Listeria monocytogenes from Genomic DNA. Journal of Clinical Microbiology. 43(7):3255-3259.

Interpretive Summary: Listeria monocytogenes is a significant cause of food-borne disease and mortality, therefore, epidemiological investigations of this pathogen require subtyping methods that are rapid, discriminatory, and reproducible. Although conventional microarray subtyping analysis (DNA probes printed on glass slides) has been shown to be both high resolution and genetically informative, it is still relatively low throughput and technically challenging. Suspension microarray technology (DNA probes attached to the surface of microscopic beads) eliminates many of the technical difficulties associated with conventional microarrays and allows high throughput subtyping of L. monocytogenes strains. In this study dendrimer signal amplification was used to increase assay sensitivity by allowing more fluorescent dye particles to attach to sample DNA and thereby increase the ratio of positive signal to background. This allowed rapid and accurate subtype identification of L. monocytogenes strains using unamplified genomic DNA. The ability to subtype genomic DNA without PCR amplification allows probes to be designed to many different regions within the bacterial genome should allow high resolution subtyping not possible with multiplex PCR.

Technical Abstract: Listeria monocytogenes is a significant cause of food-borne disease and mortality, therefore, epidemiological investigations of this pathogen require subtyping methods that are rapid, discriminatory, and reproducible. Although conventional microarray subtyping analysis has been shown to be both high resolution and genetically informative, it is still relatively low throughput and technically challenging. Suspension microarray technology eliminates the technical issues associated with planar microarrays and allows high throughput subtyping of L. monocytogenes strains. In this study, a suspension array assay using dendrimer signal amplification allowed rapid and accurate serovar identification of L. monocytogenes strains using genomic DNA as target. The ability to subtype genomic DNA without PCR amplification allows probes to be designed to many different regions within the bacterial genome should allow high resolution subtyping not possible with multiplex PCR.