Application of TaqMan low density array to simultaneous identification of multiple animal-derived components
Taking advantage of TaqMan<sup>®</sup> array microfluidic card technology, 24 primer-and-probe sets of real-time fluorescence quantitative polymerase chain reaction (RT-PCR) for identification of 24 different animal species (categories) were assembled in one array to achieve simultaneous...
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| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Zhejiang University Press
2020-04-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
| Subjects: | |
| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2019.04.221 |
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| Summary: | Taking advantage of TaqMan<sup>®</sup> array microfluidic card technology, 24 primer-and-probe sets of real-time 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 animal-derived 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 in-card 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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| ISSN: | 1008-9209 2097-5155 |