The Research For The Determination Of Turnip Mosaic Virus And Cucumber Green Mottle Mosaic Virus Using Sensors

Turnip mosaic virus is a worldwide popular kind of plant virus, which caused serious damages to the crop production. Cucumber green mottle mosaic virus is a straight baculovirus, which is the member of the genus Tobamovirus, and the main host was Cucurbitaceae. It causes leaf mottle, stunt plant growth. Therefore, establishing a rapid and reliable detection method for the prevention of the virus introduction to protect the safety of China's agricultural production is of great significance. Electrochemical immunosensor is of good selectivity, low tests cost, free sample's color and turbidity effects. The required equipment is relatively simple, fast, small, and have many other advantages. It can be widely used in medical, food analysis, industrial production, environmental monitoring and other fields. This paper developed the immune electrochemical sensors for the detection of Turnip mosaic virus and cucumber green mottle mosaic virus. The main contents are as follows:(1) The combination of enzyme-catalyzed silver deposition and signal amplification method constructs a sensitive and simple electrochemical immune sensor for detect of Turnip mosaic virus. Cysteamine and glutaraldehyde were fixed on the surface of gold electrode. Then, TuMV antibody was connected to the gold electrode surface through glutaraldehyde, eventually, the antibody was fixed to the electrode surface. The alkaline phosphatase-labeled antibody (TuMV specified antibody) was immobilized on the electrode surface through sandwich immunity reaction. The undissolving silver deposition was formed on the electrode surface through the reaction of alkaline phosphatase catalyzed ascorbic acid reducing silver ions, and then the electrochemical signal in the electrode was enlarged greatly, detected by linear sweep voltammetry (LSV). The combination of enzyme-catalyzed signal amplification and the sensitivity of electrochemical detection make the method obtain a low detection limit. The effects of experimental conditions, such as the concentration of antibody and the depositing time on the performances of immunoassay were discussed. A linear relationship between reduction wave peak current and the concentration of TuMV from 10 to 1000-fold dilution of TuMV primary solution with the detection limit of 500000-fold dilution of TuMV primary solution. This method is simple, low cost, and expected for Turnip mosaic virus detection.(2) The electrochemical bio-sensing technology based on enzyme-catalyzed and silver deposition of signal amplification to construct an electrochemical immunosensor test for cucumber green mottle mosaic virus. In this method, the alkaline phosphatase-labeled antibody (CGMMV antibody) was immobilized on the electrode surface by sandwich method, the undissolving deposition was formed on the electrode surface by using alkaline phosphatase catalyze ascorbic acid to reduce silver ions. In the end, the enzyme-catalyzed silver deposition reacted in the present substrate solution. The electrochemical signal detects through linear sweep voltammetry (LSV). The method is sensitive, simple, good selectivity and of wide linear range, low detection limit. Therefore, it is expected to have good application prospects in the CGMMV detection prevention.