Establishment of a Loop-mediated Isothermal Amplification (LAMP) Method for the Rapid Detection of Hazelnut Components in Plant-based Protein Beverages
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Abstract:
To address the widespread adulteration of plant-based protein beverages, a loop-mediated isothermal amplification (LAMP) method was developed for the rapid detection of hazelnut components. The conserved sequence (No. 19517533~19519500 bp) of chromosome Ca8 (accession number: LR899430.1) in European hazelnut was adopted to design the specific primers for the LAMP detection of hazelnut components. The reaction system was optimized and the specificity, sensitivity, and stability of the LAMP method were verified. Using real-time polymerase chain reaction as the reference method, McNemar's test was performed to verify the performance indexes and accuracy of the LAMP method for 32 plant-based protein beverages. The LAMP method had strong specificity, and no positive amplification results were obtained for 33 plant ingredients except hazelnut. The reproducibility test was stable (Ct value, RSD=5.41%), and the lower limit of detection was 0.1% (by mass fraction). The results indicated that the LAMP method had a specificity and sensitivity of 100%, with no false positives or false negatives. In conclusion, the LAMP detection method had the advantages of simple operation, fast and efficient detection (<2 h from sample processing to final results), and high accuracy. Hence, it can be applied to the rapid detection of hazelnut components in plant-based protein beverages.
