A kind of Mycoplasma bovis antibody detection reagent 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 present invention relates to a detection reagent of Mycoplasma bovis antibody and a preparation method thereof. Background technique [0002] Mycoplasma bovis is an important pathogenic pathogen that infects cattle. Hale first isolated the pathogen from the milk of mastitis in the United States in 1961. It has been confirmed that Mycoplasma bovis not only causes bovine pneumonia and mastitis, but also causes arthritis, keratoconjunctivitis, otitis, reproductive tract inflammation, miscarriage and infertility. Since 2008, an outbreak of "contagious Mycoplasma bovis pneumonia" characterized by necrotizing pneumonia has occurred in newly imported beef cattle in some areas of my country. It has caused huge economic losses to my country's cattle industry. Since then, clinical diagnosis has been reported in Gansu, Ningxia, Qinghai, Gu...

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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 technologies for detecting Mycoplasma bovis are: 1. Pathogen isolation and identification, but the isolation of Mycoplasma bovis is often affected by the clinical application of antibiotics, and the separation is difficult, and the separation medium has high nutritional requirements, the isolation is difficult to identify, 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 CaswellJL, ArchambaultM.Mycoplasmabovispneumoniaincattle); 2. PCR detection method (see ThomasA, DizierI, LindenA, etal. Disease samples are subjected to complex processing (grinding, centrifugation, DNA extraction, etc.). Although the detection method is sensitive, it is prone to false positives due to contamination and improper handling, which affects the accuracy of the judgment results. In addition, a certain laboratory is required Conditions and equipment, as well as higher costs, are not suitable for clinical or grassroots use

3. Indirect ELISA is currently the most important serological detection method. In the world, whole bacterial protein has been used to coat and express protein as a method of coating antigen (from GrandDL, CalavasD, BrankM, et al.SerologicalprevalenceofMycoplasmabovisinfectioninsucklingbeefcattleinFrance), whole bacterial protein The lack of specificity of the method is rarely used now, and the ELISA method for expressing protein antigens has been 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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