Mycoplasma hyopneumoniae antibody detection method and application

A technology for detection of Mycoplasma hyopneumoniae and antibodies, applied in chemical instruments and methods, immunoassays, biological tests, etc., can solve problems such as failure to achieve ideal results, increase reaction range and sensitivity, maintain stability, and shorten growth. effect of cycles

Pending Publication Date: 2021-07-09
中农华大(武汉)检测科技有限公司
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  • Application Information

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Problems solved by technology

In previous studies, IHA used whole bacteria protein to sensitize formaldehyde red blood cells to detect serum antibodies to My

Method used

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  • Mycoplasma hyopneumoniae antibody detection method and application
  • Mycoplasma hyopneumoniae antibody detection method and application
  • Mycoplasma hyopneumoniae antibody detection method and application

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Embodiment

[0029] The sensitizing antigens used in the indirect hemagglutination detection method for Mycoplasma hyopneumoniae in this example are P36 and P46 proteins expressed by Escherichia coli, and the concentration of the antigenic protein is 10 μg / mL. The preparation process is as follows:

[0030] 1. Preparation of Mycoplasma hyopneumoniae P36 and P46 proteins

[0031] (1) Construction and identification of expression vectors

[0032] Referring to the genome sequence of Mhp (J strain NC_007295.1), respectively design the primers for the whole gene sequence or partial gene sequence of P36 and P46 immunogenic proteins to amplify the corresponding sequences;

[0033] P36 (P36-F: GC GGATCC ATGGATAATCCTGATGCCAAACCTATTAAAATAGCTCTAATTG;

[0034] P36-R: GG AAGCTT TCATTCTCTTTTTACCTTCAATATTTTTAATTGCATCCTGATAA);

[0035] P46 (P46-F: GC GGATCC ATGATCAGTATGGAAAACAAAAAAATGCTTAGAAAAAAATTTT;

[0036] P46-R: GG AAGCTT TCATATTTTTAATAGGTTTGGCATCAGGATTATCAACATTAGCT),

[0037] PCR identif...

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Abstract

The invention discloses a detection method and application of a mycoplasma hyopneumoniae antibody. The mycoplasma hyopneumoniae antibody is detected by mixing P36 and P46 proteins expressed by escherichia coli with sensitized red blood cells. According to the method, mycoplasma hyopneumoniae specific proteins P36 and P46 are specifically selected, the indirect hemagglutination sensitization antigen prepared through escherichia coli expression and Ni column purification reacts with a mycoplasma hyopneumoniae antibody to the maximum extent, the reaction range and sensitivity are greatly increased, cross reaction with other mycoplasmas is avoided, escherichia coli expresses the sensitization protein antigen, the growth period is greatly shortened, and production cost is reduced. The method is advantaged in that the detection reagent is good in stability, high in sensitivity and good in repeatability, has no obvious cross reaction on other pathogenic antibodies, is simple, convenient and rapid to operate, and can be used for clinical sample detection, immune level monitoring and epidemiological investigation.

Description

technical field [0001] The invention relates to a method for detecting animal infectious diseases, which belongs to the field of veterinary biological products. Background technique [0002] Mycoplasma hyopneumoniae (Mhp) is the main pathogen that causes swine asthma, and the disease is widespread in the world. Local pig breeds and hybrid pig breeds in my country are particularly susceptible and almost universal, causing heavy losses. Once the pig herd is infected with Mhp, it is difficult to eliminate, and can be secondary infected with PRRSV, PCV2, etc., resulting in the failure of various vaccinations. [0003] At present, the detection techniques of mycoplasma pneumonia in swine mainly include isolation and culture techniques, PCR techniques and serological detection techniques. Separation and culture technology and PCR technology are mainly aimed at the detection and diagnosis of pathogens, which is reflected in the infection of Mhp. Due to the strict nutritional requi...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569C12N15/70C07K14/30
CPCC07K14/30C12N15/70G01N33/56933G01N33/6854G01N2469/20
Inventor 胡星星贾双冯小英丁琳林琴
Owner 中农华大(武汉)检测科技有限公司
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