摘要
本研究建立1种基于TaqMan探针的实时荧光环介导等温扩增(Loop-mediated isothermal amplification,LAMP)(TaqMan—LAMP)检测方法,用于肉制品中鸭源性成分的快速鉴别。以鸭线粒体cytb基因的保守序列设计特异性引物,优化并建立基于TaqMan探针的实时荧光LAMP检测方法,对该方法进行特异性、灵敏度试验;同时与《GB/T38164—2019常见畜禽动物源成分检测方法 实时荧光PCR法》作比较,对市售肉制品进行检测,验证本方法的准确性。所建立的TaqMan-LAMP检测方法特异性强,仅鸭肉基因组DNA呈现特异性扩增曲线,30 min内完成检测,灵敏度为0.001%(w/w),重复性好,批次内变异系数CV为1.99%,对47份肉制品进行检测,检测结果与国标法相比,相对敏感性、特异性和符合率均为100%。本研究建立的鸭源性成分TaqMan-LAMP检测方法检测快速、特异性强、灵敏度高、重复性好,适用于肉制品中鸭源性成分的快速检测。
In this study,a real-time Loop-mediated isothermal amplification assay based on TaqMan probe(TaqMan-LAMP)was developed for the rapid identification of duck-derived components in meat products.Cytb was used as the target gene to design specific primers followed by establishment and optimization of a real-time fluorescent LAMP detection method whose specificity and sensitivity was then tested and compared with GB/T38164-2019 real-time fluorescence PCR method on the commercial meat products.Only the genomic DNA of duck meat showed a specific amplification curve within 30 min with a sensitivity of 0.001%(w/w),suggesting the high specificity of the TagMan-LAMP repeatability of the method was high,and the coefficient of variation(CV)in batches was 1.99%.The detection results of 47 meat products were compared with the national standard method,and the relative sensitivity,specificity and coincidence rate were 100%.The TaqMan-LAMP detection method established in this study is rapid with strong specificity,high sensitivity and good reproducibility.Thus,the Tag-LAMP method is is suitable for rapid detection of duck-derived components in meat products.
作者
柳梦思
时国强
高娜婷
焦义然
孙旭飞
孟亚南
董振国
LIU Mengsi;SHI Guoqiang;GAO Nating;JIAO Yiran;SUN Xufei;MENG Yanan;DONG Zhenguo(Hebei sanshibio-tech co.,ltd,Shijiazhuang 050035,China;College of Food Science and Technology,Hebei Agricultural University,Baoding 071000,China;College of Life Sciences,Hebei Agricultural University,Baoding 071000,China)
出处
《河北农业大学学报》
CAS
CSCD
北大核心
2023年第6期89-94,共6页
Journal of Hebei Agricultural University
基金
国家自然科学基金项目(32172288)
石家庄市科技孵化计划项目(211510109A)。
