| 高通量基因芯片在压载水中检测致病菌的应用 |
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| 引用本文： | 张子龙,李深伟,邱瑾,田桢干,李晓虹,张继伦,卢钟山,李平.高通量基因芯片在压载水中检测致病菌的应用[J].上海海洋大学学报,2018,27(3):431-437. |
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| 作者姓名： | 张子龙 李深伟 邱瑾 田桢干 李晓虹 张继伦 卢钟山 李平 |
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| 作者单位： | 上海国际旅行卫生保健中心;上海出入境检验检疫局 |
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| 基金项目： | 国家质量基础的共性技术研究与应用专项（2016YFF0203204-2）；质检总局项目（20182K017） |
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| 摘 要： | 在口岸压载水检测中发现有多种食源性致病菌的存在,给公共卫生和口岸生态环境带来极大风险。目前对食源性致病菌的检测主要是通过细菌的培养及生化鉴定、荧光PCR等方法,但是不能同时检测多种致病菌。本研究针对一种能检测12种食源致病菌的芯片进行了应用研究,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。在DNA水平上可以检测到110~7 000 copy的DNA分子,在菌裂解液的水平上可以检测到104~105copy数目的菌体,样品添加实验证明1 cfu/m L的菌浓度就可以通过增菌后芯片检测到。该芯片可鉴别金黄色葡萄球菌、阪崎肠杆菌、肠出血性大肠杆菌O157∶H7、霍乱弧菌、志贺氏痢疾杆菌、空肠弯曲杆菌、单增李斯特菌、副溶血性弧菌、弯曲肠杆菌、沙门氏菌、志贺氏菌、蜡样芽孢杆菌和产气荚膜杆菌等。研究证明该芯片的灵敏度比PCR水平略低,但是其具备高通量、快速、准确,简单易操作等优点,可以应用于口岸压载水中携带食源致病菌的快速检测。
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| 关 键 词： | 高通量芯片 食源性致病菌 快速检测 压载水 |
| 收稿时间： | 2018/3/16 0:00:00 |
| 修稿时间： | 2018/4/15 0:00:00 |
High-throughput DNA microarray used for pathogens detection in the ballast water |
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| ZHANG Zilong,LI Shenwei,QIU Jin,TIAN Zhengan,LI Xiaohong,ZHANG Jilun,LU Zhongshan and LI Ping.High-throughput DNA microarray used for pathogens detection in the ballast water[J].Journal of Shanghai Ocean University,2018,27(3):431-437. |
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| Authors: | ZHANG Zilong LI Shenwei QIU Jin TIAN Zhengan LI Xiaohong ZHANG Jilun LU Zhongshan and LI Ping |
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| Institution: | Shanghai International Travel Healthcare Center, Shanghai 200135, China;Shanghai entry-exit inspection and quarantine bureau, Shanghai 200135, China,Shanghai International Travel Healthcare Center, Shanghai 200135, China;Shanghai entry-exit inspection and quarantine bureau, Shanghai 200135, China,Shanghai International Travel Healthcare Center, Shanghai 200135, China;Shanghai entry-exit inspection and quarantine bureau, Shanghai 200135, China,Shanghai International Travel Healthcare Center, Shanghai 200135, China;Shanghai entry-exit inspection and quarantine bureau, Shanghai 200135, China,Shanghai entry-exit inspection and quarantine bureau, Shanghai 200135, China,Shanghai entry-exit inspection and quarantine bureau, Shanghai 200135, China,Shanghai entry-exit inspection and quarantine bureau, Shanghai 200135, China and Shanghai entry-exit inspection and quarantine bureau, Shanghai 200135, China |
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| Abstract: | The potential risk of foodborne pathogens in the ballast water to act as a major vector for public health and port ecological environment has long been recognised. At present, the foodborne pathogens is detected by culture and biochemical identification, Real-time PCR and other methods, which can not detect multiple pathogenic bacteria. This study focused on the application of a high-throughput DNA microarray chip p that can detect 12 kinds of foodborne pathogens. By the chip,every control strain of pathogen can be specifically detected without inferring with other organism. In the level of DNA molecule, the chip can detect the minimal 110 copies for the most sensitive Shigella, and the maximal 7000 copies for Staphylococcus aureus. Pathogens with copies of 104-105, can be detected directly by the thermal lysis bacteria. ballast water supplied with five kinds of pathogens of 1cfu/ml can be detected in 2 hours followed with several hours of culture. the method for the simultaneous foodborne pathogens besides Staphylococcus aureus,Salmonella spp,and Shigella spp, Listeria monocytogenes, Escherichia coli O157:H7, Vibrio parahaemolyticus,Vibrio cholera, Bacillus cereus Clostridium perfringens, Campylobacter, Cronobacter sakazakii, Shigella Castellani could be detected. The minimum limit of the chip is higher than the PCR methods, while the chip of this study was high-throughput, rapid and efficient in the detection,which could be a useful method for the simultaneous detection of foodborne pathogens in the ballast water |
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| Keywords: | High-throughput DNA microarray Foodborne pathogen identificati quick detection ballast water |
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