Detection reagent for mycoplasma bovis antibody and preparation method thereof

Abstract

The invention relates to a biological detection reagent and a preparation method thereof, and particularly relates to a detection reagent for a mycoplasma bovis antibody and a preparation method thereof. The detection reagent for the mycoplasma bovis antibody is composed of one part of a mycoplasma bovis P48 recombined and expressed fusion protein antigen and one part of 10% hydroformylation-tanned sheep red blood cells (v / v) which are uniformly mixed. The gene sequence of the P48 recombined and expressed fusion protein antigen for the detection reagent for the mycoplasma bovis antibody is SEQ ID No.1. The detection reagent can be used for really detecting mycoplasma bovis and has the advantages of long preservation time, high sensitization titer, easiness in long-time preservation, high blood coagulation titer, simplicity in operation, convenience and rapidness, low cost, safety and stability, no toxin, no environmental pollution and the like.

Description

technical field [0001] The invention relates to a biological detection reagent and a preparation method of the detection reagent. Specifically, the invention relates to a detection reagent for mycoplasma bovis antibody and a preparation method thereof. Background technique [0002] Mycoplasma bovis is an important pathogenic pathogen that infects cattle. In 1961, Hale first isolated the pathogen from cow's milk with mastitis in the United States. In 1976, it was first reported to be related to bovine respiratory diseases. It has been confirmed that Mycoplasma bovis can cause other diseases such as arthritis, keratoconjunctivitis, otitis, reproductive tract inflammation, miscarriage and infertility in addition to bovine pneumonia and mastitis. Since 2008, the outbreak of "contagious mycoplasma bovis pneumonia" characterized by necrotizing pneumonia has occurred in some areas of my country newly imported beef cattle from other places. It has caused huge economic losses to my c...

AI Technical Summary

Problems solved by technology

Therefore, 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 also difficult to popularize as a routine detection method (see Caswell J L, Archambault M. Mycoplasma bovis pneumonia in cattle); 2. PCR detection method (see Thomas A, Dizier I, Linden A, et al. al .Conservation of the uvrC gene sequence in Mycoplasma bovis and its use in routine PCR diagnosis), this detection method requires complex treatment of disease samples (grinding, centrifugation, DNA extraction, etc.), although the detection method is sensitive, it is easy False positives are more likely to occur due to pollution and improper handling, which affects the accuracy of the determination 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

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