Colloidal gold chemiluminescence immune analysis method for detecting pig pleuropneumonia antibody

Abstract

The invention discloses a colloidal gold chemiluminescence immune assay method of detecting pig peripneumonia antibody. The method relates to animal virus immunodetection and chemiluminescence immune assay field. The main points of the invention are hereinafter: fixing the pig peripneumonia Apx IVA antigen in 96-microtitor plate, blocking the spare active site in the plate, add sample product to be examined, thus the pig peripneumonia Apx IVA antigen is binded. The gold-labeled rabbit anti pig IgG is added and the specific reaction with the sample can take place. The chemiluminescence reagent is added with the dissolved solution after the colloidal gold labeled on rabbit anti pig IgG is dissolved by violet acid and then detect chemiluminescence intensity. The chemiluminescence intensity is in direct ratiot to antibody concentration to be detected, thus linear correlation is available between chemiluminescence intensity and pig peripneumonia Apx IVA antibody. Compared with the prior art, the method has more accuracy and high specificity to better meet the need of clinical detection of peripneumonia.

Description

technical field [0001] The invention belongs to the technical field of immunodetection and chemiluminescence immunoassay technology, and in particular relates to a colloidal gold chemiluminescence qualitative immunoquantitative analysis method for detecting porcine pleuropneumonia antibodies. Background technique [0002] Porcine pleuropneumonia (Porcine Pleuropneumonia) is a highly contagious and fatal respiratory disease characterized by pleuropneumonia and hemorrhagic necrotizing pneumonia caused by Actinobacillus Pleuropneumoniae (APP). It is one of the five major diseases of pigs recognized in the world. Since the disease was reported in 1957, it has occurred in the United Kingdom, the United States, Germany, Canada and other countries. It is a worldwide disease and widely exists in all pig-raising countries in the world, causing huge economic losses to the pig industry. According to the clinical investigation and testing results, the disease is becoming more and more ...

AI Technical Summary

Problems solved by technology

But this patent application has the following defects: one is that the description of the application is not sufficiently disclosed, and the bacterial strain used for PCR amplification is not clearly announced

The second is that the serum dilution ratio of the application is relatively low (100 times is the best), and its sensitivity is not high.

The third is that the clinical serum samples used for the comparison test are taken from dead pigs with severe respiratory symptoms and anatomical lesions concentrated in the lungs, and the effect of early diagnosis of porcine pleuropneumonia is not obvious

Fourth, the APP capsular polysaccharide used in the comparative test method established by the invention has serotype specificity, and there may be quite a few false positives and false negatives, resulting in reduced detection reliability. According to the data reported in this field, two or There are few reports of 100% coincidence rate for more than two comparative detection methods, and the reported effect of the 100% coincidence rate claimed by the application is difficult to agree

[0007] So far, there have been no reports on methods and kits for detecting porcine pleuropneumonia antibodies using colloidal gold chemiluminescence immunoassay

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