Efficacy test method of mycoplasma gallisepticum inactivated vaccine and application thereof

A technology of Mycoplasma gallisepticum and inactivated vaccine, which is applied to measuring devices, instruments, scientific instruments, etc., can solve the problems of hidden dangers of virulent attacks on biological safety, errors in test results, easy to disperse poison, etc., and achieves small batch differences and results. Accurate, repeatable results

Inactive Publication Date: 2013-05-08
兆丰华生物科技(南京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above-mentioned experimental evaluation results often have errors in the test results due to the deviation of subjective factors, and there are also biological security risks in strong poison attacks, and it is easy to disperse the poison.

Method used

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  • Efficacy test method of mycoplasma gallisepticum inactivated vaccine and application thereof
  • Efficacy test method of mycoplasma gallisepticum inactivated vaccine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Efficacy Testing Method of Mycoplasma Gallisepticum Inactivated Vaccine

[0022] 1. Vaccination

[0023] 20 40-day-old SPF chickens were used, 10 of which were injected subcutaneously at the back of the neck or intramuscularly with thighs to immunize 1 portion, and the other 10 were not immunized as controls. After 30 days, blood was collected, serum was separated, and ELISA antibody level or HI antibody titer was determined.

[0024] 2. Mycoplasma gallisepticum enzyme-linked immunosorbent assay (ELISA) and hemagglutination inhibition (HI) test

[0025] 1. Mycoplasma gallisepticum ELISA test

[0026] Purchase MG antibody ELISA (ELISA, MG Ab) kit: produced by IDEXX Company in the United States (reagents include: coated microplate, sample diluent, positive control, negative control, enzyme-labeled secondary antibody, substrate solution and stop solution) , Beijing IDEAS Century Yuanheng Co., Ltd.

[0027] The ELISA detection method was carried out according to the IDE...

Embodiment 2

[0053]Parallel relationship between serum antibody titer and challenge protection rate of mycoplasma gallisepticum inactivated vaccine immunization test animals Use 3 batches of vaccines, each batch divides 60 40-day-old SPF chickens into 6 groups, 10 in each group, 50 of them were immunized by subcutaneous injection of 20 μl, 50 μl, 200 μl, 300 μl, 500 μl of the back of the neck or intramuscularly of the thigh, and the other 10 were not immunized as a control. After 30 days, blood was collected, serum was separated, and ELISA antibody level or HI antibody level was determined. At the same time, together with 10 control chickens, at 2-3m 3 Spray and challenge Mycoplasma gallisepticum R strain culture 500-600mL (each 1.0ml contains viable bacteria count 108-9CCU color change units) in a closed room (indoor temperature is 15°C-30°C, relative humidity is 50%-70%), The duration of spraying should not be less than 5 minutes, and the droplet size should be around 2.0 μm. Observe fo...

Embodiment 3

[0058] Comparison of serum antibody and challenge protection rate of Mycoplasma gallisepticum inactivated vaccine single vaccine and combined vaccine immunization experimental animals Three batches of vaccines were used for each single vaccine of Mycoplasma gallisepticum inactivated vaccine and combined vaccine of Mycoplasma gallisepticum inactivated vaccine. Ten 40-day-old SPF chickens were immunized with the same antigenic content of Mycoplasma gallisepticum inactivated vaccine single vaccine and Mycoplasma gallisepticum inactivated vaccine combined vaccine, and immunized with subcutaneous or thigh intramuscular injection at 0.2ml / feather, and another 10 Only non-immunization was used as a control. After 30 days, blood was collected, serum was separated, and ELISA antibody level or HI antibody level was determined. At the same time, together with 10 control chickens, at 2-3m 3 Spray and challenge Mycoplasma gallisepticum R strain culture 500-600mL (each 1.0ml contains 108-9...

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Abstract

The invention relates to the technical field of animal biological products and discloses an efficacy test method of mycoplasma gallisepticum inactivated vaccine. Simultaneously, the efficacy test method disclosed by the invention can also be applied to quality control of the vaccine, in particular comprising the determination of immune toxicity attack protection rate and vaccine quality standard, measurement of immunization deadline and clinical monitoring. Experiments prove that the animal toxicity attack test is replaced by a serology detection method (comprising ELISA (Enzyme-Linked Immuno Sorbent Assay) and HI (Hemagglutination Inbition)); the efficacy test method is simple, convenient, rapid, accurate in result and good in repeatability and specificity; due to the establishment of the efficacy test method, subjectivity for checking airbag pathological change integrals after attacking toxicity is reduced; toxicity dispersion is avoided; a basis for establishing a rational immune program is also provided; and the efficacy test method has general popularization significance.

Description

technical field [0001] The invention relates to a method for testing the efficacy of mycoplasma gallisepticum inactivated vaccine and an application thereof, belonging to the field of biotechnology. Background technique [0002] Mycoplasma gallisepticum is one of the most common pathogens of poultry. The disease often spreads slowly in large-scale chicken farms, and the main losses caused to the chicken industry are the obvious increase in the death rate of infected chickens; the development of broiler chickens and growing chickens is hindered, and the feed remuneration is reduced; the egg production of laying hens is reduced. ; With the improvement of the intensification degree of my country's chicken industry, the prevalence of Mycoplasma gallisepticum is becoming more and more violent, and secondary infections are caused by other diseases. In recent years, there are often mixed infections of respiratory diseases that cause a large number of chickens to die, and the frequen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
Inventor 丁美娟王伟尹秀凤黄显明许秀梅孙汪爱芬吴娟张海涛徐延伟薛家宾张小飞
Owner 兆丰华生物科技(南京)有限公司
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