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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2012, Vol. 32 Issue (6): 84-92    
技术与方法     
九种单核细胞增生性李斯特菌检测技术效果比较及评价
刘雅莉1,2, 刘芳3, 韩舜愈1, 刘箐2, 孙志博4, 夏俊芳1, 朱小清5
1. 甘肃农业大学食品科学与工程学院 兰州 730070;
2. 上海理工大学医疗器械与食品学院 上海 200093;
3. 甘肃出入境检验检疫局国际旅行卫生保健中心 兰州 730020;
4. 兰州大学基础医学院 兰州 730000;
5. 甘肃出入境检验检疫局 兰州 730020
Comparison and Evaluation of Nine Methods in Detecting Listeria monocytogenes
LIU Ya-li1,2, LIU Fang3, HAN Shun-yu1, LIU Qing2, SUN Zhi-bo4, XIA Jun-fang1, ZHU Xiao-qing5
1. College of Food Science and Engineering, Gansu Agricultural University, Lanzhou 730070, China;
2. School of medical instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China;
3. Internation Travel Healthcare Center of Gansu Entry-Exit Inspection and Quarantine Bureau, Lanzhou 730020, China;
4. School of Basic Medical Sciences, Lanzhou University, Lanzhou 730030, China;
5. Gansu Entry-Exit Inspection and Quarantine Bureau, Lanzhou 730020, China
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摘要： 采用传统生物学方法、显色培养基方法、自制三抗体夹心ELISA试剂盒(TAS-ELISA)、基因组直接PCR(DNA Direct-PCR)、菌裂解直接PCR(Bacterium Direct-PCR)、免疫捕捉PCR(IC-PCR)、生物PCR(Bio-PCR)、SYBR Green染料实时荧光PCR(SYBR Green Real time-PCR)、探针实时荧光PCR(TaqMan Real time-PCR)检测纯菌液及模拟带菌食品中的单核细胞增生性李斯特菌(Listeria monocytogenes, LM)。结果表明:传统生物学培养方法出现假阳性;TAS-ELISA、DNA Direct-PCR、Bacterium Direct-PCR、IC-PCR最低检测限分别为106、102、105、104CFU/ml;显色培养基、SYBR Green Real time-PCR灵敏度达102CFU/ml;Bio-PCR、TaqMan Real time-PCR检测灵敏度最高,均为101CFU/ml。显色培养基、TAS-ELISA操作简单,适合大量样品的初检;IC-PCR具有灵敏、特异、快速、经济的优点,特别适合大体积样品中微量病原的检测;Bio-PCR、TaqMan Real time-PCR灵敏度高、特异性好,适合阳性样品的复检以及科研使用。
关键词： 单核细胞增生性李斯特菌;  三抗体夹心ELISA;  免疫捕捉PCR;  生物PCR;  实时荧光定量PCR    
Abstract: Nine methods were used to detect Listeria monocytogenes in bacterial suspensions and artificial contaminated food extracts, including traditional biological methods, chromogenic medium method, TAS-ELISA, DNA Direct-PCR, Bacterium Direct-PCR, IC-PCR, Bio-PCR, SYBR Green Real time-PCR and TaqMan Real time-PCR, and the results were analyzed and compared. The result showed that Traditional biological methods had false positive. The lowest detectable limits of TAS-ELISA, DNA Direct-PCR, Bacterium Direct-PCR and IC-PCR were 106, 102, 105, 104CFU/ml. The detection sensitivity of chromogenic medium method, SYBR Green Real time-PCR was 102CFU/ml. Bio-PCR, TaqMan Real time-PCR had the highest sensitivity which could reach 101CFU/ml. Chromogenic medium and TAS-ELISA had the advantages of simple operation, and they were suitable for preliminary screening of large samples. IC-PCR was a sensitive, specific, rapid, economic technology, suitable for detecting micro-pathogens in large volume samples particularly. Bio-PCR and TaqMan Real time-PCR had high sensitivity and specificity, which adapted to recheck and research.
Key words: Listeria monocytogenes    TAS-ELISA    IC-PCR    Bio-PCR    Real time-PCR
收稿日期: 2012-02-13 出版日期: 2012-06-25
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刘雅莉
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夏俊芳
朱小清

引用本文:

刘雅莉, 刘芳, 韩舜愈, 刘箐, 孙志博, 夏俊芳, 朱小清. 九种单核细胞增生性李斯特菌检测技术效果比较及评价[J]. 中国生物工程杂志, 2012, 32(6): 84-92.

LIU Ya-li, LIU Fang, HAN Shun-yu, LIU Qing, SUN Zhi-bo, XIA Jun-fang, ZHU Xiao-qing. Comparison and Evaluation of Nine Methods in Detecting Listeria monocytogenes. China Biotechnology, 2012, 32(6): 84-92.

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https://manu60.magtech.com.cn/biotech/CN/    或    https://manu60.magtech.com.cn/biotech/CN/Y2012/V32/I6/84

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