Rapid Detection Of Escherichia Coli O157:H7 On Food Safety

E.coli O157: H7(Escherichia coli O157:H7)is a serious hazard of foodborne bacterial pathogens which can infected diarrhea,hemorrhagic colitis,also can cause serious complications such as hemolytic uremic syndrome and even lead to death.E.coli O157:H7 has become a global public health problem due to its characteristics such as outbreak trend,strong pathogenicity and mortality.The enterohemorrhagic E.coli as one of the pathogenic microorganisms were classified in China,and it may have a significant impact on the health.Because traditional bacteriological identification is cumbersome and time-consuming,it is not conducive to early identification.In recent years,various simple,rapid and accurate examination methods have been published.Rapid and specific detection method on E.coli O157:H7 is essential for early diagnosed of the disease.In this paper,the E.coli O157:H7 was identified in many aspects of microorganism,the method of multiple polymerase chain reaction(PCR)was established and applied,the real samples were also analyzed.First,in order to understand the basic biological characteristics of E.coli O157:H7,the E.coli O157:H7 was cultivated and tested.The E.coli O157:H7 was preliminarily identified by the SMAC media.Secondly,the three primers were designed by the specific gene stx1,stx2 and specificity gene flagella flic.The multiple PCR reaction system was established.The results show that all three pairs of primers can be effectively detected and it has good specificity.By optimizing experimental conditions as follows:the annealing temperature 57?,the primer concentration was 1?mol/L,DNA template was 3 ?L.It can be detected in 5 hours to find out whether the sample contains E.coli O157:H7.With the gradual improvement of people's food safety awareness,establishing a rapid and specific assay method have practical implications.Farmers market was a important place for food circulation and is most closely connected with lives of people.Therefore,By using the established method,Sixty pork samples were collected form Wuhan farmers markets,only one positive result was detected.The factors that cause positive result were analyzed.This method can meet the demand of massive determination,and it also has the advantages of low cost,high speed,high specific and high accuracy.