The Visual Detection Methods Of Staphylococcus Aureus By Using Click Chemistry Technology

Staphylococcus aureus is widely distributed in living environment.It can pollute meat,aquatic products,dairy products and animal feed,and cause people or animals serious vomiting,diarrhea clinical symptoms,and bring huge economic losses.Traditional detection methods of Staphylococcus aureus was costly,time-consuming,cumber and can not control pollution foods.We need to establish Staphylococcus aureus sensitive and convenient detection methods to improve food hygiene standards to protect people's lives and property.This study combine with click chemistry,HCR,nano-flower and other visual signal output platforms,this study optimized the reaction conditions such as the best probe concentration and the optimal incubation time in the reaction system,and carried out specific and sensitive double evaluation,the establishment of sensitive and specific detection of Staphylococcus aureus specific test methods,the specific experimental results follows:(1)Test strip with copper free click chemistry for visual of Staphylococcus aureusIn this study,the copper-free chemistry and test strip combine to establish the visualization detection method of Staphylococcus aureus.The optimum concentration of CCLCR was 50 n M and the best connection temperature was 25?.It can be specifically detect Staphylococcus aureus target sequence,the minimum detection limit of 10 f M,the milk as a simulated sample,detection recovery rate of 86% to 90%,through the naked eye can observe the results,in order to achieve the actual sample visual inspection provides a new foundation.(2)Visual detection Staphylococcus aureus by microplate combined with HCRIn this study,microtiter plate chromatography and HCR technology combine to establish a method for visualization of Staphylococcus aureus.The optimal screening time is 0.5 ?M,the best incubation time is 2 h,and the HCR product was not modified.The optimal incubation time of the HCR product is 2 h.The designed detection model could distinguish the sequence of a base difference with a minimum detection limit of 32.5 f M.The minimum detection limit of Staphylococcus aureus is 52.1 cfu/ml.(3)Nanoflowers combined with copper click chemistry visual of Staphylococcus aureusThe nanoflowers particles and copper click chemistry combine to establish the visualization method decetion of Staphylococcus aureus.Through the optimizing experimental conditions,the optimal antibody concentration is 5 mg/ml,best incubation time is 2 h,and concentration is 25 m M.The limit detection of Staphylococcus aureus can reach 102 cfu /ml.In summary,this study combine with click chemistry,HCR,nanoflowers and other visual signal output platform,the establishment of three visual detection methods for Staphylococcus aureus,through the three detection methods for specificity,sensitivity testing and evaluation to finally ensure that the detection of Staphylococcus aureus,the successful realization of the visual detection of Staphylococcus aureus,and its operation at room temperature,the test results easy to judge,without precision instruments,reducing the cost of detection etc.It can increase detection and provided a good direction for development POCT detection of foodborne pathogens.