The application relates to the field of biological detection, and specifically discloses a sealing agent for biological detection, a preparation method thereof, a coated plate and a kit using the coated plate. Each liter of sealing agent is made of the following raw materials: 2.5-9g of water-soluble alcoholic organic matter, 0-1.1g of polyglucosamine, 0-0.7g of gelatin, 0.1-0.4g of antioxidant, 0.07-0.2g of preservative, and the balance It is water; the water-soluble alcoholic organic matter is selected from at least two of ethylene glycol, polyvinyl alcohol and polyethylene glycol. The preparation method of the sealing agent comprises the following steps: uniformly mixing water-soluble alcoholic organic matter, polyglucosamine, gelatin, antioxidant, preservative and water according to the formula quantity. The coated plate is prepared after being sealed with the above sealing agent, and the kit includes the above prepared coated plate. The use of the blocking agent of the present application can effectively reduce the non-specific reaction in the detection process, with few false positive results and high diagnostic specificity.