Abstract:

Objective To establish a new rapid screening method for detection of E.sakazakii—cross-primer amplification combined with immuno-blotting analysis. Method On the basis of sequence analysis of the E. sakazakii 16S-23S rDNA internal transcribed spacer,specific primers and probe were designed, we developed cross-primer isothermal amplification method, using the immuno-gold standard test strip to test results. The specificity of the method was evaluated by 18 strains of E.sakazakii and 36 negative strains of Enterobacteriaceae.The sensitivity of the method was evaluated by testing the quantitative DNA,pure bacteria counted and the sample of adding other bacterial. Result The method established in this article is highly specific and sensitive. The limit of decetion is 100fg/test for the genomic DNA, 101 cfu/mL for pure bacterial culture. Conclusion The method is highly sensitive,specific, easy to operate and does not require any complex equipment. It is very useful for detection of E. sakazakii in the port or underdeveloped areas.