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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  2018, Vol. 38 Issue (12): 91-98    DOI: 10.13523/j.cb.20181212
综述     
纳米孔测序技术应用于食品微生物检测的可行性分析
曹必溥1,缪丽华1,郭宝颜1,贺丽苹2,**()
1 广州广检质量检测研究院有限公司 广州 510000
2 华南农业大学测试中心 广州 510642
The Feasibility Analysis That Nanopore Sequencing Technology Is Applied to Food Microbiological Detection
CAO Bi-pu1,MIAO Li-hua1,GUO Bao-yan1,HE Li-ping2,**()
1 Guangzhou Guangjian Quality Testing and Research Institute Co.Ltd.,Guangzhou 510000,China
2 Testing Center of South China Agricultural University,Guangzhou 510642,China
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摘要：

近些年来DNA测序技术发展迅速,已经从第一代生化测序发展到第三代单分子测序。作为第三代测序技术中的一种不同于当前流行的其他测序技术,纳米孔测序技术是基于电信号的一种物理方法测序。许多研究者通常将高通量测序技术应用于食品微生物的研究,但是将纳米孔测序技术应用于食品中微生物的检测却鲜有报道。Oxford Nanopore Technologies(牛津纳米孔科技公司)研发的DNA测序仪MinION,是世界首例用于商业测序的纳米孔测序仪,经过不断完善,近年来MinION在DNA测序中被广泛应用。MinION 测序一次需要的DNA量约1μg,其标准识别速度为一秒钟识别250个碱基,平均读长可至13kb~20kb,测序准确率可以达到98%。纳米孔测序的高识别速度和高准确率,完全满足快速检测的要求,将其应用于食品中微生物检测是完全可行的。
关键词： 纳米孔测序;  DNA测序技术;  食品微生物    
Abstract:

DNA sequencing technology has developed rapidly in recent years and has evolved from the first generation of biochemical sequencing to the third generation of single molecule sequencing. As one of the third-generation sequencing technologies, nanopore sequencing is a physical method that based on electrical signals,different from other current popular sequencing technologies. Many researchers usually apply high-throughput sequencing techniques to the study of food microorganisms, but there are few reports of the application of nanopore sequencing technology to the detection of microorganisms in food. Oxford Nanopore Technologies successfully developed DNA sequencing instrument of MinION, the world’s first commercial instrument for the nanopore sequencing. After continuous improvement in recent years, MinION has been widely used in the DNA sequencing. The amount of DNA required for the sequencing of MinION is about 1μg DNA. The standard recognition speed of one second for 250 bases, the average reading length of 13kb to 20kb and sequencing accuracy to 98%. Its high recognition speed and high accuracy meet the requirements of rapid detection, feasible to be applied to microbiological detection in food.
Key words: Nanopore sequencing    DNA sequencing technology    Food microorganism
收稿日期: 2018-05-04 出版日期: 2019-01-10
ZTFLH:  Q523  
通讯作者: 贺丽苹     E-mail: 582262409@qq.com
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引用本文:

曹必溥,缪丽华,郭宝颜,贺丽苹. 纳米孔测序技术应用于食品微生物检测的可行性分析[J]. 中国生物工程杂志, 2018, 38(12): 91-98.

CAO Bi-pu,MIAO Li-hua,GUO Bao-yan,HE Li-ping. The Feasibility Analysis That Nanopore Sequencing Technology Is Applied to Food Microbiological Detection. China Biotechnology, 2018, 38(12): 91-98.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20181212    或    https://manu60.magtech.com.cn/biotech/CN/Y2018/V38/I12/91

图1  生物纳米孔测序原理图[31]
图2  α-溶血素(a)和耻垢分枝杆菌孔蛋白A(MspA) (b)三维模型[33]
图3  α-溶血素与phi29 DNA聚合酶的测序组合体[36]
图4  MspA与phi29 DNA 聚合酶的测序组合体[39]
图5  MinION测序仪[25]
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