Development of a DNA macroarray for simultaneous detection of multiple foodborne pathogenic bacteria in fresh chicken meat

Abstract

A DNA macroarray was developed to provide the ability to detect multiple foodborne pathogens in fresh chicken meat. Probes targeted to the 16S rRNA and genus- and species-specific genes, including fimY, ipaH, prfA, and uspA, were selected for the specific detection of Salmonella spp., Shigella spp., Listeria monocytogenes, and Escherichia coli, respectively. The combination of target gene amplification by PCR and a DNA macroarray in our system was able to distinguish all target bacteria from pure cultures with a detection sensitivity of 10⁵ c.f.u. ml⁻¹. The DNA macroarray was also applied to 10 fresh chicken meat samples. The assay validation demonstrated that by combining the enrichment steps for the target bacteria and the DNA macroarray, all 4 target bacteria could be detected simultaneously from the fresh chicken samples. The sensitivity of L. monocytogenes and Shigella boydii detection in the fresh chicken samples was at least 10 and 3 c.f.u. of the initial contamination in 25 g samples, respectively. The advantages of our developed protocol are high accuracy and time reduction when compared to conventional culture. The macroarray developed in our investigation was cost effective compared to modern oligonucleotide microarray techniques because there was no expensive equipment required for the detection of multiple foodborne pathogens.

The Use of Macroarray as a Simple Tool to Follow the Metabolic Profile of <i>Lactobacillus plantarum</i> during Fermentation

“…Macroarray technology has been successfully applied also e.g. for the detection of pathogens in chicken samples [35] and studies on environmental samples for the presence of specific antibiotic resistance genes [36] communities of diazotrophs [37], and expression of 375 genes in L. lactis subsp. lactis IL1403 during stress conditions [38].…”

Prevalence and Characterization of Pathogenic Bacteria in Bulk Tank Raw Milk, Thailand

“…The prevalence of bacterial isolates in local area containing the virulence genes were measured by using PCR technique. The genomic DNA (gDNA) of target bacterial isolates was extracted from 16-24 h grown in pure cultures on TSA using phenol-chloroform methods [9]. The gDNA template of each isolate was amplified by PCR technique using gene specific primers as shown in Table 1.…”