大肠杆菌特异性快速检测系统研制.pdf

第23卷第7期
传感技术学报
2010年7月
CHINESE JOURNAL OF SENSORS AND ACTUATORS
July 2010
Development of an Instrument for Specific and Rapid Detection of Escherichia Coli
TIAN Qing. 2, LIU Ruping 2, LIU Xiahong, LUO Jinping, Catxinaia
State Key Laboratory of Transducer echnology, Institute of Electronics, Chinese Academy of Sciences, Beijing 100190, China
2. Graduate School of Chinese Academy of Sciences, Bejjing 100190, China
Abstract: Combination of adenosinase triphosphate(ATP)bioluminescence principles and immune magnetic sepa-
ration technology, an instrument for specific rapid detection of bacteria is presented, which can achieve high speci-
ficity of bacteria rapid detection sensitivity. By improving light shielding design of optical detecting module and
noise suppression design of amplification and A/D ciruit, strict controlling of reaction conditions, the detection
background was reduced. so as to achieve a lower detection limit and improve the detection sensitivity. E. coli
(ATCC25922 )standard solutions were tested in the experiment by the system, and the detection limit reached
18CFU/ml. Compared with the result obtained by the standard aerobic plate count APC method, the correlation co-
efficient (r) is 0. 993. And the reproducibility of the system was tested. The relative average deviation(R.A. D
is 6.48% and the coefficient of variability(CV) is 9.33 %. Combined with immunomagnetic separation tech-
niques, E. coli solutions mixed with high concentrations of Staphylococcus aureous(ATCC25923)were tested
The linear correlation coefficient between the results detected by the system and the APC method is 0.957 1, indi
cating that this apparatus and method has a good specificity. The system is rapid, specific, accurate and repetitive
which can be used in rapid detection of pathogens in food and environment.
Key words rapid detection system; specific; bioluminescence; adenosine triphosphate(ATP); immunomagnetic sepa
ration
EEACC:7210:4190:1200
大肠杄菌特异性快速检测系统研制
田青,刘儒平2,刘晓红2,罗金平,蔡新霞2
中国科学院电子学研究所传感技术联合国家重点实验室,北京100190
2.中国科学院研究生院,北京100190
摘要:结合三磷酸腺苷(ATP)生物发光原理和免疫磁分离技术,研制了一种细菌特异性快速检测系统,可实现对细菌的特
异性高灵敏度快速检测。该仪器通过改进光学检测模块的避光设计和放大采集电路的噪声抑制设计,及严格控制试剂反应
条件,降低了检测背景,从而达到了更低的检测下限并提高了检测灵敏度。对大肠肝菌(ATCC25922)标准菌溶液进行检测,
检测下限达到18CFU/mL,与培养计数法检测结果相关系数R达到0.93,选取一份样品进行重复性检测,相对平均偏差(R
D.)为6.48%,变异系数(CV)为9.33%。结合免疫磁分离技术,对混入的高浓度金黄色葡萄球菌的大肠杆菌溶液进行检
测,检测结果与原始大肠肝菌培养计数结果一致,证明该仪器及方法对检测大肠肝菌具有良好的特异性。该系统检测速度
快,准确度较高,重复性良好,特异性好,可用于食品及环境中的致病菌快速检测。
关键词:快速检测系统;特异性;生物发光;三磷酸腺苷(ATP);免疫磁分离
中图分类号:TH79
文献标识码:A文章编号:1004-1699(2010)07-0899-04
细菌特异性检测在食品卫生安全、水环境检测、速检测的需求。流式细胞计术法、聚合酶链反
医疗卫生等领域都有着重要应用,但目前常用的平应法(PCR)4、电化学阻抗谱法S-?、生物发光
板计数方法需要至少24h的培养,不能满足现场快法等方法都被用于细菌快速检测。其中生物发
项目来源:国家863项目资助(NO2009AA037411);国家科技重大专项资助(NO2008710003003-02);中科院知识创新工程项
目资助;中科院优秀奖项目资助
收稿日期:2010-01-08修改日期:2010-02-24