Development and comparison of two assay formats for parallel detection of four biothreat pathogens by using suspension microarrays.

PloS One
Ingmar JanseBart J van Rotterdam

Abstract

Microarrays provide a powerful analytical tool for the simultaneous detection of multiple pathogens. We developed diagnostic suspension microarrays for sensitive and specific detection of the biothreat pathogens Bacillus anthracis, Yersinia pestis, Francisella tularensis and Coxiella burnetii. Two assay chemistries for amplification and labeling were developed, one method using direct hybridization and the other using target-specific primer extension, combined with hybridization to universal arrays. Asymmetric PCR products for both assay chemistries were produced by using a multiplex asymmetric PCR amplifying 16 DNA signatures (16-plex). The performances of both assay chemistries were compared and their advantages and disadvantages are discussed. The developed microarrays detected multiple signature sequences and an internal control which made it possible to confidently identify the targeted pathogens and assess their virulence potential. The microarrays were highly specific and detected various strains of the targeted pathogens. Detection limits for the different pathogen signatures were similar or slightly higher compared to real-time PCR. Probit analysis showed that even a few genomic copies could be detected with 95% confid...Continue Reading

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Citations

May 20, 2014·Biosensors & Bioelectronics·Subash C B GopinathThangavel Lakshmipriya
Jun 3, 2014·Acta Veterinaria Scandinavica·Gernot SchmoockLisa D Sprague
Oct 25, 2013·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Jane Christopher-HenningsJoan K Lunney

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Datasets Mentioned

BETA
IS1111
CP000915
only

Methods Mentioned

BETA
PCR

Software Mentioned

Visual Oligonucleotide Modeling Platform
SPSS Statistics
Kodon
BLAST
Insignia
Visual OMP

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