Wang Xin-Wei, Zhang Liang, Jin Lian-Qun, Jin Min, Shen Zhi-Qiang, An Shuang, Chao Fu-Huan, Li Jun-Wen
Institute of Environment and Health, No. 1, Dali Road, Tianjin, 300050, People's Republic of China.
Appl Microbiol Biotechnol. 2007 Aug;76(1):225-33. doi: 10.1007/s00253-007-0993-x. Epub 2007 May 10.
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.
食源致病菌的快速准确检测与鉴定对食品安全至关重要。在本文中，我们描述了一种快速（<4小时）的高通量检测与鉴定系统，该系统使用通用聚合酶链反应（PCR）引物扩增细菌16S rRNA基因的可变区，然后将产物与芯片上的物种特异性寡核苷酸探针进行反向杂交。该方法成功区分了来自纯培养物的属于13个细菌属的204株细菌。当将该方法直接应用于从食品中分离的115株细菌时，112/115（97.4%）被正确鉴定；两株由于信号弱无法区分，而一株未能产生PCR产物。该芯片用于检测并成功鉴定了来自食物中毒爆发样本的两株细菌，杂交结果与传统方法相同。微阵列分析的灵敏度为10(2) CFU细菌。因此，寡核苷酸微阵列是检测和鉴定食品中病原体的有力工具。
