• EI
  • Scopus
  • 食品科学与工程领域高质量科技期刊分级目录第一方阵T1
  • DOAJ
  • EBSCO
  • 北大核心期刊
  • 中国核心学术期刊RCCSE
  • JST China
  • FSTA
  • 中国精品科技期刊
  • 中国农业核心期刊
  • CA
  • WJCI
  • 中国科技核心期刊CSTPCD
  • 中国生物医学SinoMed
中国精品科技期刊2020
柳海宾, 张海鑫, 宋腾飞, 刘聪, 聂付磊. 环介导等温扩增结合免疫层析试纸条快速检测羊肉及其制品中的鸭源成分[J]. 食品工业科技, 2020, 41(16): 238-242,251. DOI: 10.13386/j.issn1002-0306.2020.16.038
引用本文: 柳海宾, 张海鑫, 宋腾飞, 刘聪, 聂付磊. 环介导等温扩增结合免疫层析试纸条快速检测羊肉及其制品中的鸭源成分[J]. 食品工业科技, 2020, 41(16): 238-242,251. DOI: 10.13386/j.issn1002-0306.2020.16.038
LIU Hai-bin, ZHANG Hai-xin, SONG Teng-fei, LIU Cong, NIE Fu-lei. Rapid Detection of Duck Derived Components in Mutton and Mutton Products by Loop Mediated Isothermal Amplification Combined with Immunochromatographic Test Strip[J]. Science and Technology of Food Industry, 2020, 41(16): 238-242,251. DOI: 10.13386/j.issn1002-0306.2020.16.038
Citation: LIU Hai-bin, ZHANG Hai-xin, SONG Teng-fei, LIU Cong, NIE Fu-lei. Rapid Detection of Duck Derived Components in Mutton and Mutton Products by Loop Mediated Isothermal Amplification Combined with Immunochromatographic Test Strip[J]. Science and Technology of Food Industry, 2020, 41(16): 238-242,251. DOI: 10.13386/j.issn1002-0306.2020.16.038

环介导等温扩增结合免疫层析试纸条快速检测羊肉及其制品中的鸭源成分

Rapid Detection of Duck Derived Components in Mutton and Mutton Products by Loop Mediated Isothermal Amplification Combined with Immunochromatographic Test Strip

  • 摘要: 为实现肉及肉制品中掺假成分的快速检测,本文将环介导等温扩增技术(loop mediated isothermal amplification,LAMP)与免疫层析试纸条(immunochromatographic test strip,ICTS)相结合,建立了一套可在羊肉及其制品中特异并灵敏地检测出鸭源性成分的方法。根据鸭特异性细胞色素b(cytb)基因的6个特异性区域设计LAMP引物,其中两条内引物FIP和BIP分别使用生物素(biotin)和地高辛(digoxin)标记。使用LAMP扩增技术于65℃下扩增30 min产生生物素和地高辛标记的双链DNA产物,将扩增产生的双链DNA产物(10 μL)与90 μL上样缓冲液(PBS pH=7.4)混合均匀后使用免疫层析试纸条进行可视化检测。本研究所建立的LAMP-ICTS方法能够在40 min内特异性地检测出羊肉及其制品中的鸭源成分,且与其他肉类不存在交叉反应,检出限可达到0.01%(w/w)。市售实际样品检测结果显示,所采集的羊肉片、羊肉串样品中均有鸭源性成分,且此结果与行业标准方法(PCR-电泳)的检测结果一致。LAMP-ICTS方法具有灵敏度高、特异性好、检测速度快、对仪器的依赖低等优点,适用于基层监管部门对羊肉及其制品真伪进行快速检测。

     

    Abstract: In order to realize the rapid detection of adulteration in meat and meat products,this paper combined loop mediated isothermal amplification(LAMP)with immunochromatographic test strip(ICTS)to establish a specific and sensitive method for detecting duck meat in mutton and its products. LAMP primers were designed according to six specific regions of duck specific cytochrome b(cytb)gene. Two inner primers,FIP and BIP,were labeled with biotin and digoxin respectively,and amplified at 65℃ for 30 min to produce double-strand DNA produces labeled with biotin and digoxin. The products(10 μL)was mixed with 90 μL loading buffer(PBS pH=7.4),and then the ICTS was used to realize visual detection. The LAMP-ICTS method established in this study could specifically detect duck ingredients within 40 min,and there was no cross reaction with other meats. At the same time,the detection limit of the target in meat products could reach 0.01%(w/w). The test results of commercial samples showed that there were duck-derived components in the samples of mutton slices and mutton kettles,and the test results were consistent with the industry standard method(PCR-electrophoresis). Therefore,LAMP-ICTS method had the advantages of high sensitivity,good specificity,rapid detection,low dependence on instruments,and was suitable for the rapid detection of meat adulteration in the laboratories of both regulatory agencies and commercial services.

     

/

返回文章
返回