Chicken infectious bronchitis live vaccine potency detection method

A bronchitis and chicken infectivity technology, applied in the field of vaccine efficacy detection, chicken infectious bronchitis live vaccine efficacy detection field, to achieve the effect of reduced requirements, strong specificity and uniform size

Active Publication Date: 2015-06-10
RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to overcome the defects of existing efficacy detection methods, and provide a detection method for measuring the efficacy of chicken infectious bronchitis live vaccine by combining PCR and chicken embryo infection method, so as to improve the accuracy of efficacy detection

Method used

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  • Chicken infectious bronchitis live vaccine potency detection method
  • Chicken infectious bronchitis live vaccine potency detection method
  • Chicken infectious bronchitis live vaccine potency detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Contrast test of potency detection of chicken infectious bronchitis live vaccine (H120 strain)

[0070] 1. Materials

[0071] Chicken infectious bronchitis live vaccine (strain H120): produced by Ruipu (Baoding) Biopharmaceutical Co., Ltd., 1000 pigeons / bottle, batch number 140504.

[0072] 2. Method

[0073] (1) Virus dilution inoculation and culture

[0074] Dilute the vaccine with sterile saline to 1 feather / 0.1ml according to the feather portion indicated on the bottle label, and then make a 10-fold serial dilution. take 10 -3 、10 -4 、10 -5 Three dilutions were used to inoculate 10-day-old SPF chicken embryos into the allantoic cavity, 5 eggs were inoculated for each dilution, 0.1ml per embryo, and 2 groups were inoculated in parallel at the same dilution, and a negative control of normal saline was set at 37°C. Continue to incubate. 24 hours after inoculation, eggs were illuminated once, and dead embryos were discarded.

[0075] (2) Virus Harvesting

[00...

Embodiment 2

[0107] 3 pairs of primers for chicken infectious bronchitis live vaccine (H120 strain) efficacy detection comparison test

[0108] 1. The materials are the same as in Example 1.

[0109] 2. Method:

[0110] According to the method of the first group in Example 1, the allantoic fluid was aseptically collected 72 hours after inoculation for detection. The total RNA of the same allantoic fluid was detected by RT-PCR with primer pair 1, primer pair 2, and primer pair 3 respectively.

[0111] 3. Test results

[0112] (1) The detection result of primer pair 1 is the same as that in Example 1; the PCR gel electrophoresis diagram of primer pair 2 is shown in image 3 ; The PCR gel electrophoresis of primer pair 3 is shown in Figure 4 .

[0113] (2) The titer determination results of the three pairs of primers are shown in Table 4.

[0114] Table 4 Statistical results of titer determination of 3 pairs of primers

[0115] .

[0116] 4 Conclusion

[0117] The method of the pr...

Embodiment 3

[0119] Contrast test of potency detection of chicken infectious bronchitis live vaccine (H52 strain)

[0120] Live chicken infectious bronchitis vaccine (strain H52): produced by Ruipu (Baoding) Biopharmaceutical Co., Ltd., 1,000 birds / bottle.

[0121] With the method of Examples 1 and 2, the potency of chicken infectious bronchitis live vaccine (H52 strain) was determined simultaneously with the standard chicken embryo infection method and the method of the present invention (3 pairs of primers). The detection results of primer pair 1 and primer pair 3 were consistent with those of chicken embryo infection method, all of which were 10 4.3 EID 50 / feather portion; primer pair 2 detection titer is 0, can not be used for the mensuration of chicken infectious bronchitis virus H52 strain titer.

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Abstract

The invention relates to a chicken infectious bronchitis live vaccine potency detection method, which comprises the following steps: 1) designing a primer; 2) diluting, inoculating and cultivating virus; 3) harvesting virus; 4) performing PCR detection; and 5) calculating virulent valence. The chicken infectious bronchitis live vaccine potency detection method is simple to operate, uniform in determination conditions, strong in controllability, small in batch errors, the detection period of the chicken infectious bronchitis virus potency is shortened, and the application range is wide.

Description

technical field [0001] The invention relates to a vaccine efficacy detection method, in particular to a chicken infectious bronchitis live vaccine efficacy detection method, which belongs to the field of biotechnology. Background technique [0002] Chicken infectious bronchitis (Infectious Bronchitis, IB) is an acute, highly contagious infectious disease of chickens caused by Infectious Bronchitis Virus (IBV) of the Coronaviridae family. caused huge losses. At present, vaccination is an effective measure to prevent and treat the disease. Therefore, the potency test (potency test) of the semi-finished venom of chicken infectious bronchitis live vaccine and the finished vaccine is very important. [0003] At present, the standard method for testing the potency of chicken infectious bronchitis live vaccines is the chicken embryo infection method (half the egg infection dose, EID 50 ), "The Veterinary Pharmacopoeia of the People's Republic of China" (Part Three, 2010 Edition)...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/686C12Q1/70C12Q2600/16
Inventor 张丽青梁武郁宏伟刘涛柳珊刘超朱秀同杨保收邹立宏
Owner RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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