Development of Rapid Screen Method of Phosphomannose Isomerase

A loop mediated isothermal amplification(LAMP) assay was developed for the rapid detection of gene PMI.A set of sepcific primers,two inner primers and outer primers 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,1.6 mmol/L.The reaction time and temperatures were optimized for 40 min at 62.0 ℃.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.