Detection of Yersinia enterocolitica in alfalfa, mung bean, cilantro, and mamey sapote (Pouteria sapota) food matrices using DNA microarray chip hybridization.
Four different food matrices (alfalfa, cilantro, mamey sapote, and mung bean) were contaminated with three different dilutions 10(6), 10(4), and 10(3) cfu/g of Yersinia enterocolitica. DNA was isolated from each food mix and used in chromosomal amplifications. The amplified DNA was used as templates in single PCR reactions of the four genes (virF, ail, yst, and blaA) followed by mixing the four reactions for one PCR primer extension reaction. The presence and the limit of detection of four genes in four food matrices were established by microarray hybridization. Data revealed the diversity of signal intensities. Neither the microarray chip hybridization nor the single PCR amplification could detect virF's presence located on a plasmid. Ail was detected in 10(3) cfu/g, whereas blaA and yst were detected from 10(5) to 10(6) cfu/g in all food matrices. Therefore, the ail gene could be the gene of choice in identifying Y. enterocolitica in alfalfa, cilantro, mamey, and mung bean. Other genes--blaA, yst, virF--exhibited wide variability in hybridization signals, highlighting the need of a better DNA purification step prior to DNA microarray hybridization.
Siddique
Nusrat
N
Department of Cell Biology and Molecular Genetics, School of Life Science, University of Maryland, College Park, MD, USA.
Sharma
Devang
D
Al-Khaldi
Sufian F
SF
D015169:Colony Count, Microbial; D028526:Coriandrum / Q000382:microbiology; D007887:Fabaceae / Q000382:microbiology; D005516:Food Microbiology; D005798:Genes, Bacterial; D031252:Mammea / Q000382:microbiology; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D015008:Yersinia enterocolitica / Q000235:genetics / Q000302:isolation & purification*