Kit for detecting avian onfectious synovitis antibody and preparation method

Abstract

The invention provides an ELISA kit for detecting avian onfectious synovitis antibody. The kit adopts the ELISA kit for detecting the avian onfectious synovitis antibody, based on avian onfectious synovitis pyruvate kinase (PYK) recombinant protein as envelope antigen. The invention further provides a preparation method of the envelope antigen of the kit, namely PYK protein. The PYK protein prepared through the method has excellent reactogenicity, and the ELISA detection kit provides an effective method for the diagnosis of avian onfectious synovitis and the detection of an antibody level after immunization, and has an obvious promotion effect on the prevention research on avian onfectious synovitis.

Description

technical field [0001] The invention relates to a bacterium kit and a preparation method thereof, in particular to an ELISA kit for detecting antibodies to Mycoplasma aviosa. Background technique [0002] Mycoplasma (Mycoplasma) is a class of microorganisms without cell walls and precursors, with few organelles, highly pleomorphic, and the smallest prokaryotic cell type that can grow and reproduce in non-living media. Mycoplasma avium mainly involves poultry, and is widely parasitic in the respiratory tract, cloaca, digestive tract, oviduct mucosa and joint capsule of poultry. At present, there are 29 species of Mycoplasma avium identified and named, of which 25 species belong to the genus Mycoplasma, 3 species of Cholesterogenum and 1 species of Ureaplasma. Among these mycoplasma, Mycoplasma gallisepticum (Mycoplasma gallisepticum, MG) and Mycoplasma synoviae (Mycoplasma synoviae, MS) are mainly mycoplasma that can infect chickens and turkeys and cause disease. However, c...

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Problems solved by technology

[0003] The current detection methods for MS include: isolation and culture of mycoplasma, serological antibody detection and molecular gene diagnosis, among which isolation and culture is the gold standard for diagnosis of mycoplasma infectious diseases, but mycoplasma has high nutritional requirements on the medium, grows slowly, and is difficult to separate , only applicable to laboratory testing; serological testing such as plate agglutination test (SPA) has low specificity, while hemagglutination inhibition test (HI) has low sensitivity and requires multiple sampling, which is more suitable for population-level detection, and the results are accurate The difference is closely related to the source of the antigenic strain; polymerase chain reaction (PCR) and real-time fluorescent quantitative PCR (real-time PCR) are currently the most widely used diagnostic methods, but there are also problems of many steps, high cost, and time-consuming ( At least 4.5 hours), it is not easy to operate on site and promote to the grassroots; currently foreign commercial Mycoplasma synovialum indirect ELISA kits are mainly from the United States and Europe, although the specificity is high, the sensitivity of antibody detection for different strains of infection is not enough, In particular, the price is expensive, and the cost of testing a blood sample is as high as 11.5 yuan, which limits the use of farmers and is limited to laboratory research.

At present, there is still no development of cheap, specific, and highly sensitive kits that use highly expressed recombinant proteins as coated antigens for ELISA detection.

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