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Pig mycoplasma specific monoclonal antibody and its application in clinical detection

A monoclonal antibody, Mycoplasma suis technology, applied in measuring devices, scientific instruments, instruments, etc., can solve the problems of long process, laborious and time-consuming, inaccurate diagnosis methods, and achieve simple operation, high titer, and strong specificity. Effect

Inactive Publication Date: 2017-06-09
深圳金石邦信投资咨询有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] So far, the diagnosis of swine mycoplasma disease in China is still based on the analysis of epidemiology, observation and analysis of clinical symptoms, and observation of pathological anatomy. Compared with the original diagnosis method, the process is longer, labor-intensive and time-consuming, and it is very imprecise.

Method used

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  • Pig mycoplasma specific monoclonal antibody and its application in clinical detection
  • Pig mycoplasma specific monoclonal antibody and its application in clinical detection
  • Pig mycoplasma specific monoclonal antibody and its application in clinical detection

Examples

Experimental program
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Effect test

preparation example Construction

[0049] (1) Preparation of immune spleen cells

[0050] ① Take the spleen of the immunized mouse, put it into a plate containing 5ml of incomplete culture medium (1640 culture medium without calf serum), put it on a 200-mesh stainless steel mesh, and squeeze all the spleen cells into the solution through the mesh.

[0051] ②Transfer the spleen cell suspension into a 50ml centrifuge tube, add incomplete culture medium to 30ml, mix well, centrifuge at 1000rpm for 5 minutes, and discard the supernatant.

[0052] ③Pour 30ml of incomplete culture solution into the centrifuge tube with precipitated splenocytes, shake gently to make the splenocytes become suspension again, then centrifuge at 1000rpm for 5 minutes, and discard the supernatant. Suspend the precipitated cells to 10ml with incomplete culture medium, mix well, count with a cell counter, and adjust the total cell number to 1x10 8 .

[0053] (2) Preparation of SP20 mouse myeloma cell suspension

[0054] ① SP20 mouse myeloma...

Embodiment 1

[0107] Example 1. A pig farm in Chang'an County, Shaanxi Province raised 500 pigs. Recently, it was found that individual pigs did not eat well, and asked for help in detecting whether there was Mycoplasma hyopneumoniae. We randomly took 10 cases of pig oral mucus, and tested them with this immunoassay kit, all of which were negative. It shows that the whole group of pigs does not have mycoplasma hyopneumoniae disease, and there may be other reasons for the poor eating of individual pigs.

[0108] ELISA reaction result table

[0109] Sample (oral mucus) Detection method Test result (positive+, negative-) 1 ELISA - 2 ELISA - 3 ELISA - 4 ELISA - 5 ELISA - 6 ELISA - 7 ELISA - 8 ELISA - 9 ELISA - 10 ELISA -

Embodiment 2

[0110] Example 2, another pig farm in Chang'an County raised 300 pigs, including 8 breeding sows, 2 boars, 50 piglets and 240 fattening pigs. Recently, some sows have been found to be coughing and eating poorly. Ask for help in testing for Mycoplasma hyopneumoniae. We randomly selected 10 cases of oral mucus and tested them with this kit. The results showed that all 3 sows tested were positive for Mycoplasma suis; 2 cases of 3 piglets were positive for Mycoplasma suis; 3 cases of 4 fattening pigs were positive. It is positive for Mycoplasma suis, indicating that the pigs have been infected with Mycoplasma suis, and measures should be taken to prevent and control it.

[0111] ELISA reaction result table

[0112] Sample (oral mucus) Detection method Test result (positive+, negative-) 1 sow ELISA + 2 sows ELISA + 3 sows ELISA + 4 piglets ELISA + 5 piglets ELISA + 6 piglets ELISA - 7 fattening pigs ELISA + ...

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Abstract

The invention relates to the field of an animal biological product, and particularly discloses a pig mycoplasma specific monoclonal antibody and its application in clinical detection; the pig mycoplasma specific monoclonal antibody is prepared by using mycoplasma hyopneumoniae as immunogen; a a pig mycoplasma specific monoclonal antibody hybridoma cell strain 2G2 with high stable secretion effect and strong specificity is successfully screened, the generated monoclonal antibody can generate immunoreaction with mycoplasma pneumonia and ureaplasma urealyticum at the same time; a pig mycoplasmosis detection kit is set up. The pig mycoplasma specific monoclonal antibody with strong specialty and high effect is used for preparing the pig mycoplasmosis immunologic diagnosis kit; the kit is sensitive, exact, and simple in operation, needs not to collect blood as sample, and not harms animals; the kit can quickly and exactly diagnose pig mycoplasma in early period; the kit can detect both of mycoplasmosis and ureaplasma urealyticum, thereby largely reducing cost and improving the detection effect.

Description

technical field [0001] The invention relates to the field of veterinary biological products, in particular to a specific monoclonal antibody of Mycoplasma suis and its application in clinical testing. Background technique [0002] More and more studies have shown that Mycoplasma suis is one of the important pathogens that cause pig respiratory tract and genitourinary tract diseases. Most of the pigs in the world are susceptible to Mycoplasma suis, and pigs of any age will be affected; according to reports The infection rate of breeding pigs in the United States is 99%, the infection rate of breeding pigs in Britain is 70%, the infection rate of breeding pigs in France is 60%, the infection rate of breeding pigs in Belgium and Canada is 80%, and the infection rate of breeding pigs in Germany and Holland is 50%. The worldwide average infection rate of farmed pigs is 50%-80%. The typical symptoms of mycoplasma hyopneumoniae disease are cough, dyspnea, and panting in sick pigs,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/577G01N33/531
CPCG01N33/56933G01N33/531G01N33/577
Inventor 黄威权赵锋吕葆真
Owner 深圳金石邦信投资咨询有限公司
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