Indirect ELISA (Enzyme-Linked Immunosorbent Assay) method and kit for detecting haemophilus parasuis antibodies

Abstract

The invention discloses indirect ELISA (Enzyme-Linked Immunosorbent Assay) method and kit for detecting haemophilus parasuis antibodies. The indirect ELISA method comprises the steps of: taking a haemophilus parasuis OMPP5 protein as an envelope antigen; wherein the judgment standard is as followed: OD450 value of to-be-detected serum is greater than 0.375, OD450 value of the to-be-detected serum / OD450 value of standard negative serum is greater than or equal to 2.1. The kit for realizing the method comprises an envelope buffer solution, confining liquid, a washing buffer solution, an antibody diluent, a chromogenic substrate solution and a stop solution, wherein the envelop antigen used in the method is the haemophilus parasuis OMPP5 protein. Proved by a specificity test, a blocking-up experiment, a repeatability test and clinic application detection, the invention has the characteristics of excellent specificity, high sensitiveness, good repeatability, rapidness, simpleness and accuracy, and can be used in clinic large-scale detection of haemophilus parasuis antibodies and epidemiological survey.

Description

technical field [0001] The invention relates to the technical field of molecular biology, animal bacteriology and animal infectious disease detection, in particular to an indirect ELISA method and a kit for detecting Haemophilus parasuis antibodies. Background technique [0002] Haemophilus parasuis disease is one of the important causes of morbidity and mortality in weaned piglets. Haemophilus parasuis (HPS), as a common bacterium in the upper respiratory tract of pigs, invades the body under certain conditions and can cause severe systemic diseases (Glasser's disease), such as fibrinous polyserositis, joint inflammation and meningitis are the main features. This disease has been considered to be a sporadic, stress-induced disease in piglets in the past. However, in recent years, with the development of pig farming industry, more and more high-health pig herds or SPF pig herds appear. If these pigs are in an unimmunized state, once HPS invades, they will be seriously infe...

AI Technical Summary

Problems solved by technology

However, the PCR and Southern hybridization methods based on the detection of its DNA are difficult to apply in practice, and CF also has the disadvantages of many components involved in the reaction, complex influencing factors, and cumbersome operation steps. IHA and ELISA methods are currently the main HPS methods explored by scholars at home and abroad. Antibody detection method, the difference is the preparation method of the antigen, and Feng Xiaoming (2009), Shi Bi (2007) etc. use ultrasonic breakage antigen and capsular polysaccharide as the establishment of the indirect ELISA method of coating antigen. The result proves that the sensitivity of ELISA Sex is higher than IHA

At present, lipopolysaccharide, LPS, and inactivated whole bacteria are commonly used as coating antigens for ELISA at home and abroad. The test results are not stable and often have false negatives, or the coincidence rate and sensitivity have not been verified.

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