Rapid Detection of Synthetic α-Amylase Gene Amy 797E Using Loop-mediated Isothermal Amplification Method

In this study,a loop mediated isothermal amplification(LAMP) assay was developed for the rapid detection of gene PMI.A set of sepcific primers,two primers,two inner and outer primers and a loop primer,were designed.The concentrations: FIP and BIP,both 0.8 μmol/L,F3 and B3,both 0.2 μmol/L,Mg2+,2 mmol/L,Betaine,1.4 mmol/L.The reaction time and temperatures were optimized for 40 min at 62 ℃.SYBR Green I detection method can be finished in 2 h.Electrophoresis detection method can be finished in 2.5 h.The LAMP assay has a higher specificity compared with Real Time PCR and conventional PCR.The detection limit of GM elements reaches 0.01 %.The results indicated the suitability and simplicity of the test as a rapid method for PMI specific detection.