Mycoplasma bovis detection test strip and preparation method thereof

Abstract

The invention discloses a mycoplasma bovis antibody detection test strip and a preparation method thereof. The test strip provided by the invention comprises a bottom plate and a sample absorption pad, a colloidal gold pad coated with an antigen, a nitrocellulose membrane and a water absorption pad which are all arrayed on the bottom plate in sequence, wherein the nitrocellulose membrane is coated with a quality control line and a detection line; the detection line is coated with a Goat Anti-Bovine Secondary antibody IgG; the quality control line is a rabbit derived P48 fusion protein. The test strip provided by the invention can detect mycoplasma bovis antibody, including blood serum antibody and milk serum antibody, and has the advantages that the test strip is low in cost, simple to operate, convenient and quick to detect, requires no special equipment and apparatus, is high in detection susceptibility, strong in specificity, wide in application range and the like.

Description

technical field [0001] The invention relates to a method for preparing a biological detection reagent and the reagent. Specifically, the invention relates to a method for preparing a test strip for detecting mycoplasma bovis antibody and the prepared test strip. Background technique [0002] Mycoplasma bovis is a serious pathogenic pathogen that can cause various diseases such as bovine respiratory disease, dairy cow mastitis, arthritis, keratoconjunctivitis, reproductive tract inflammation, abortion and infertility, and can cause other pathogenic diseases once infected Secondary infections such as Cryptobacterium pyogenes, Pasteurella multocida, Mannella hemolytica, Haemophilus somniferus, bovine parainfluenza virus type 3, herpesvirus type 1, bovine respiratory syncytial virus, bovine viral diarrhea virus and bovine coronavirus, etc. The incidence rate of Mycoplasma bovis is 50%-100%, and the fatality rate is as high as 10%-50%, which has caused huge economic losses to m...

AI Technical Summary

Problems solved by technology

The actual prevalence of this disease in my country and accurate epidemiological information are still relatively scarce. The main reason for this phenomenon is that my country currently lacks relevant technologies for effective detection of Mycoplasma bovis.

[0003] The existing techniques for detecting Mycoplasma bovis are: 1. Isolation and identification of pathogens. However, the isolation of Mycoplasma bovis is often affected by the clinical application of antibiotics, and the separation is difficult. The medium for separation requires high nutrition, the identification of the isolate is difficult, and the sensitivity is low. As a method for early and rapid diagnosis of Mycoplasma bovis infection, it is difficult to popularize as a routine detection method (from Caswell J L, Archambault M. Mycoplasma bovis pneumonia in cattle); 2. PCR detection method (from Thomas A, Dizier I, Linden A, et al .Conservation of the uvrC gene sequence in Mycoplasma bovis and its use in routine PCR diagnosis), this detection method requires complex processing of disease samples (grinding, centrifugation, DNA extraction, etc.), although the detection method is sensitive, but False positives are more likely to occur due to pollution and improper handling, which will affect the accuracy of the judgment results. In addition, certain laboratory conditions and equipment are required, as well as high cost. It is not suitable for clinical or grassroots use.

3. Indirect ELISA is currently the most important serological detection method. In the world, whole bacterial protein coating and expressed protein have been used as the coating antigen method (from Grand D L, Calavas D, Brank M, et al. Serological prevalence of Mycoplasma bovis infection in suckling beef cattle in France), the lack of specificity of the whole bacteria protein method has been rarely used, and the expression protein antigen ELISA method is currently commercialized, but which antigen protein it uses has not been announced

In addition, the ELISA method is only suitable for testing in the laboratory, and the operation steps are relatively complicated, requiring professionals to operate, and certain laboratory conditions and equipment are required for detection and analysis of results

4. The indirect hemagglutination detection method established by using P48 recombinant expression protein coated sheep red blood cells is relatively simple to operate, but it also takes 2 to 3 hours to determine the result. The sensitivity is weak and a specific constant temperature incubator is required.

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