| 免疫磁珠捕获PCR快速检测单核细胞增生李斯特氏菌 |
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| 引用本文： | 李婧姮,郭慧琴,高晓强,刘 芳,邱 实,李建武,曾海娟,刘 兰,宋春美,刘 箐.免疫磁珠捕获PCR快速检测单核细胞增生李斯特氏菌[J].食品科学,2015,36(12):226-229. |
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| 作者姓名： | 李婧姮 郭慧琴 高晓强 刘 芳 邱 实 李建武 曾海娟 刘 兰 宋春美 刘 箐 |
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| 作者单位： | 1.上海理工大学医疗器械与食品学院，上海 200093；2.上海慧耘生物科技有限公司，上海 200433；;
3.甘肃出入境检验检疫局，甘肃 兰州 730020 |
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| 基金项目： | 上海理工大学大学生卓越工程项目；上海市科委重点支撑项目（13430502400） |
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| 摘 要： | 利用免疫磁珠富集单核细胞增生李斯特氏菌,采用聚合酶链式反应(polymerase chain reaction,PCR)扩增进行快速检测。所制备的免疫磁珠选取在37℃条件下均匀振荡1 h为最佳包被条件,1 mg磁珠偶联抗体的最佳量为100μg/mg,制备的免疫磁珠捕获率为45%。所建立的免疫磁珠捕获-PCR技术在样品细菌浓度达到104 CFU/m L即可被检出,其灵敏度是直接PCR检测限(105 CFU/m L)的10倍,可为病原菌的富集和快速检测提供新方法。
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| 关 键 词： | 免疫磁珠 单核细胞增生李斯特氏菌 聚合酶链式反应 |
Immunomagnetic Capture PCR for Rapid Detection of Listeria monocytogenes |
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| LI Jingheng,GUO Huiqin,GAO Xiaoqiang,LIU Fang,QIU Shi,LI Jianwu,ZENG Haijuan,LIU Lan,SONG Chunmei,LIU Qing.Immunomagnetic Capture PCR for Rapid Detection of Listeria monocytogenes[J].Food Science,2015,36(12):226-229. |
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| Authors: | LI Jingheng GUO Huiqin GAO Xiaoqiang LIU Fang QIU Shi LI Jianwu ZENG Haijuan LIU Lan SONG Chunmei LIU Qing |
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| Affiliation: | 1. College of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology,;
Shanghai 200093, China; 2. Prajna Biology Technique Co. Ltd., Shanghai 200433, China;
3. Gansu Entry-Exit Inspection and Quarantine Bureau, Lanzhou 730020, China |
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| Abstract: | This experiment was conducted using immunomagnetic enrichment followed by amplification with polymerase
chain reaction (PCR) for rapid detection of Listeria monocytogenes. The best coating conditions were achieved by shaking
the prepared immunomagnetic beads at 37 ℃ for 1 h, and the amount of antibody conjugated to the immunomagnetic beads
was 100 μg/mg, leading to a capture rate of 45%. The immunomagnetic capture PCR assay was able to detect as low as
104 CFU/mL Listeria monocytogenes with 10 times higher sensitivity than direct PCR showing a limit of detection of
105 CFU/mL. This PCR assay can provide a new method for the enrichment and rapid detection of pathogenic bacteria. |
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| Keywords: | IMBs Listeria monocytogenes PCR |
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