Development and application of an oligonucleotide microarray for the detection of food-borne bacterial pathogens.
The rapid and accurate detection and identification of food-borne pathogenic bacteria is critical for food safety. In this paper, we describe a rapid (<4 h) high-throughput detection and identification system that uses universal polymerase chain reaction (PCR) primers to amplify a variable region of bacterial the 16S rRNA gene, followed by reverse hybridization of the products to species-specific oligonucleotide probes on a chip. This procedure was successful in discriminating 204 strains of bacteria from pure culture belonging to 13 genera of bacteria. When this method was applied directly to 115 strains of bacteria isolated from foods, 112/115 (97.4%) were correctly identified; two strains were indistinguishable due to weak signal, while one failed to produce a PCR product. The array was used to detect and successfully identify two strains of bacteria from food poisoning outbreak samples, giving results through hybridization that were identical to those obtained by traditional methods. The sensitivity of the microarray assay was 10(2) CFU of bacteria. Thus, the oligonucleotide microarray is a powerful tool for the detection and identification of pathogens from foods.
Wang
Xin-Wei
XW
Institute of Environment and Health, No. 1, Dali Road, Tianjin, 300050, People's Republic of China.
Zhang
Liang
L
Jin
Lian-Qun
LQ
Jin
Min
M
Shen
Zhi-Qiang
ZQ
An
Shuang
S
Chao
Fu-Huan
FH
Li
Jun-Wen
JW
D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D017931:DNA Primers / Q000235:genetics*; D005504:Food Analysis; D005516:Food Microbiology; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D013045:Species Specificity; D014774:Virulence
D017931:DNA Primers; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S