摘要
研制了1种高通量检测食品中常见致病微生物的寡核苷酸微阵列芯片。该芯片在种的水平可鉴别金黄色葡萄球菌、阪崎肠杆菌、肠出血性大肠埃希氏菌0157:H7、霍乱弧菌、空肠弯曲杆菌、大肠弯曲杆菌、单核细胞增生李斯特菌、乙型溶血型链球菌和副溶血性弧菌;在属的水平可鉴别弯曲肠杆菌属、沙门氏菌属和李斯特氏菌属。研究中利用16S rDNA和沙门氏菌、O157:H7和副溶血性弧菌的特异基因序列设计引物和探针。用参比菌株和实验室自分离的菌株检测该芯片的特异性,结果表明该芯片的特异性良好,在所检测的菌株之间无交叉反应,与同属的其他菌株之间也不存在交叉反应。在鲜活海产品的24h增菌液中添加参比菌株,当样品增菌液中的目的菌的量达到10^6~10^7 CFU/mL时,芯片可以准确检测出目的菌。
We developed oligonucletide microarrays to detect pathogenic bacteria commonly found in food. Staphylococcus aureus, Enterobacter sakazakii, E. coli O157 : H7, Vibrio cholera, Campylobacter spp. , Campylobacter jejuni, Campylobacter coli, Salmonella spp. , Listeria spp. , Listeria monocytogenes, Vibrio parahaemolyticus and Streptococcus pyogenes can be detected by our microarrays simultaneously. Bacterial 16S ribosomal DNA was used to design PCR primers and probes except for E. coli O157: H7, Salmonella spp. and Vibrio parahaemolyticus which use species-specific primers and probes. The array can successfully identify 119 stains with no ambiguous hybridization. When adding certain amount of reference stains to 24h enrichment of seafood, the array can detect reference stain only when the target stain's concentration is up to 10^6~10^7 CFU/mL.
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2007年第12期122-126,共5页
Food and Fermentation Industries
基金
国家质检总局科技支撑计划(项目编号:2003IK049)