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DNA Microarrays

DNA Microarrays

Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...

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Updated: Mar 3, 2026

Colorimetric Paper-based Detection of Escherichia coli, Salmonella spp., and Listeria monocytogenes from Large Volumes of Agricultural Water

Colorimetric Paper-based Detection of Escherichia coli, Salmonella spp., and Listeria monocytogenes from Large Volumes of Agricultural Water

Published on: June 9, 2014

Diagnostic microarray for 14 water and foodborne pathogens using a flatbed scanner.

Vidya Srinivasan¹, Robert D Stedtfeld¹, Dieter M Tourlousse¹

¹Department of Civil and Environmental Engineering, Michigan State University, East Lansing, MI 48824, United States.

Journal of Microbiological Methods

|April 26, 2017

Summary

This summary is machine-generated.

Researchers developed a cost-effective microarray for simultaneous detection of 14 bacterial pathogens. This method uses gold labeling with silver enhancement (GLS) and multiplex polymerase chain reaction (PCR) for reliable, high-throughput screening.

Keywords:

Gold labeling and silver enhancement Low cost microarray Virulence and marker genes Waterborne pathogens

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Area of Science:

Microbiology
Biotechnology
Molecular Diagnostics

Background:

Simultaneous detection of multiple water and foodborne pathogens is crucial for public health.
Limited resources in laboratories necessitate cost-effective and accessible diagnostic approaches.
Existing methods may not offer the required throughput or affordability for widespread use.

Purpose of the Study:

To develop and validate a high-density microarray for simultaneous screening of 14 bacterial pathogens.
To establish a cost-effective and reliable method for pathogen detection suitable for resource-limited settings.
To optimize probe design and data analysis for accurate presence/absence calls.

Main Methods:

Designed 8887 50-mer probes targeting virulence and marker genes (VMGs) using an in-house database.
Synthesized probes in quadruplicate on glass slides using in-situ synthesis technology.
Generated target VMG amplicons via multiplex polymerase chain reaction (PCR), labeled with biotin, and hybridized to the microarray.
Employed gold labeling with silver enhancement (GLS) protocol for signal generation.
Quantified signals using a high-resolution flatbed scanner.

Main Results:

Established reliable presence/absence call criteria: >4 probes/gene, signal-to-noise ratio (SNR) cutoff ≥2, and positive fraction (PF) >0.75.
Achieved 100% correct calls with no false positives in blind sample testing.
Demonstrated sensitivity comparable to standard PCR due to multiplex PCR amplification.
Validated the cost-effectiveness and reliability of the GLS microarray approach.

Conclusions:

The developed high-density microarray offers an inexpensive and reliable technique for high-throughput screening of multiple bacterial pathogens.
This GLS-based approach is suitable for laboratories with limited resources, enhancing pathogen surveillance capabilities.
The method provides accurate and sensitive detection, comparable to PCR, for simultaneous identification of various pathogens.