Position: Home > Articles > Application of TaqMan low density array to simultaneous identification of multiple animal-derived components
Journal of Zhejiang University (Agriculture and Life Sciences)
2020,46
(2)
189-200
利用TaqMan微流控阵列同步检测多种动物源性成分
作 者：
虞惠贞;侯寒黎;尹文秀;张荃;孙超;张明哲;张晓峰;苗丽;吴姗
单 位：
浙江省检验检疫科学技术研究院
关键词：
TaqMan微流控阵列;动物源性成分;同步检测
摘 要：
利用TaqMan微流控阵列(TaqMan~? array microfluidic card)技术,将24种(类)动物物种的实时荧光定量聚合酶链式反应(real-time fluorescence quantitative polymerase chain reaction, RT-PCR)的引物探针集合在同一个阵列上,以实现多物种高通量同步检测,并对该方法的重复性、特异性和灵敏度进行验证。用变异系数(coefficient of variation, CV)表示一块阵列板内,同一物种成分检测的重复性。结果显示:66个CV值中,11个低于1%,15个大于2%,其余40个介于1%～2%之间,没有一个物种成分的CV值高于3%。P值显示的是不同阵列板间同一成分检测的重复性,22个物种成分的P值只有1个小于0.05,显示为板间存在差异,其他21种成分的板间差异均未达到显著水平。因此,本研究对多数物种成分的检测,不论板内还是板间都有较好的重复性。特异性检测中未发现TaqMan微流控阵列中出现假阳性的结果,但有2种成分的检测出现了假阴性。TaqMan微流控阵列的检测灵敏度没有高于单重荧光定量PCR,在22个检测样品中,10个物种成分检测的灵敏度降为原来的10%,1个甚至降低为原来的1%,其余11个灵敏度和单重荧光保持一致。总体而言,TaqMan微流控阵列技术在重复性、特异性和灵敏度方面都能满足物种成分真伪鉴定的需求,为同步鉴定多种动物源性成分提供了一种新的方法。
译 名：
Application of TaqMan low density array to simultaneous identification of multiple animal-derived components
作 者：
YU Huizhen;HOU Hanli;YIN Wenxiu;ZHANG Quan;SUN Chao;ZHANG Mingzhe;ZHANG Xiaofeng;MIAO Li;WU Shan;Zhejiang Academy of Science and Technology for Inspection and Quarantine;Qianjiang Customs;Technology Center of Zhengzhou Customs;
关键词：
TaqMan low density array;;animal-derived components;;simultaneous identification
摘 要：
Taking advantage of TaqMan~? array microfluidic card technology, 24 primer-and-probe sets of realtime fluorescence quantitative polymerase chain reaction(RT-PCR) for identification of 24 different animal species(categories) were assembled in one array to achieve simultaneous detection of multiple animal-derived components. The repeatability, specificity and sensitivity of the method were verified. Coefficient of variation(CV) value was used to indicate the repeatability in one array card when detecting the same component(species).Among 66 CV values, 11 CV values were less than 1%; 15 CV values were more than 2%; and the remaining 40 CV values were between 1% and 2%; but none of them was higher than 3%. P value showed the repeatability among different array cards when detecting the same component. Among the 22 P values of the 22 animalderived components, only one was less than 0.05, which indicated the significant difference of the CV values of the components between the cards. But none of the resting 21 components showed significant inter-plate differences. Therefore, the vast majority of the components tested in this study showed good repeatability both incard and inter-card. No false positive results were found in TaqMan array tests, but false negative results were appeared in two component detection. The sensitivity of TaqMan array assay was not higher than that of the individual RT-PCR assay. Ten of the 22 components decreased to 10% of the originals in detection sensitivities;another one component even decreased to 1%. And the remaining 11 components were consistent with that of the individual RT-PCR. However, in general, TaqMan array approach can meet the needs of authenticity identification in terms of repeatability, specificity and sensitivity, which offers promise for rapid and simultaneous identification of multiple animal-derived components.
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