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Pseudorabies antibody blocking test paper

A technology of pseudorabies and detection test strips, which is applied in the field of pseudorabies antibody blocking detection test strips, can solve the problems of ineffective recognition of monoclonal antibodies, and achieve the effects of broad market prospects, high labeling rate, and simple operation

Inactive Publication Date: 2019-07-30
河南百奥生物工程有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although blocking ELISA can still specifically detect the newly emerging mutated PRV virus antibodies in my country, the possibility of invalid recognition of monoclonal antibodies still exists

Method used

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  • Pseudorabies antibody blocking test paper
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  • Pseudorabies antibody blocking test paper

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0056] (1) Preparation of pseudorabies virus immune antigen

[0057] (1.1) Preparation of pseudorabies virus gE recombinant protein

[0058] The epitope-enriched region of PRV gE protein was amplified by PCR, and the target gene was cloned into the prokaryotic expression vector pET-28a to construct the gE gene recombinant expression plasmid pET-gE, which was induced by 0.4mol / L IPTG and highly expressed in Escherichia coli The gE recombinant protein and Western blot detection showed that the gE expressed protein could be specifically recognized by the virulent PRV positive serum and had good reactivity. The gE recombinant protein (12mg / L) of purity about 90% was obtained by Ni column affinity chromatography purification technology ( figure 1 ).

[0059] (1.2) Preparation of pseudorabies virus gB recombinant protein

[0060] Amplify the neutralizing epitope-enriched region of PRV gB protein by PCR, clone the target gene into the prokaryotic expression vector pET-28a, and con...

Embodiment 1

[0128] Example 1, Antibody monitoring of pseudorabies infection, collect animal serum, take 100 μL serum sample and add 300 μL PBS or normal saline to dilute, prepare serum solution to be tested, use pseudorabies antibody blocking detection test paper to perform detection and results according to (15) operation method To judge, detect the antibody level of pseudorabies infection. The test paper shows three red bands "|||", and the color intensity of the detection line T1 and T2 is equivalent to that of the blank control, which is negative for pseudorabies antibody, indicating that the sample to be tested does not contain pseudorabies antibody; three red bands appear "| ||", but the color intensity of the detection line T1 and T2 is significantly weaker than that of the blank control, which is weakly positive for pseudorabies infection antibody, indicating that the sample to be tested contains a low level of infection antibody; only a red band "|" appears, which is a pseudorabie...

Embodiment 2

[0129] Example 2, Evaluation of the immunization effect of pseudorabies vaccine, 2 to 3 weeks after immunization with inactivated pseudorabies vaccine, collect serum from immunized animals, take 100 μL serum samples and add 300 μL PBS or normal saline for 1:2, 1:4, 1:8 …Double ratio dilution, prepare the serum solution to be tested, use the pseudorabies antibody blocking detection test paper to perform detection and result judgment according to the operation method (15), and detect the level of pseudorabies immune antibody. The test paper shows three red bands "|||", and the color intensity of the detection line T1 and T2 is equivalent to that of the blank control, which is negative for pseudorabies antibody, indicating that the sample to be tested does not contain pseudorabies antibody; three red bands appear "| ||", and the color intensity of the detection line T2 is significantly weaker than that of the blank control, which is weakly positive for pseudorabies immune antibody...

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Abstract

The invention discloses pseudorabies antibody blocking test paper, composed of a supporting plate, an antigen pad, a gold labeling pad, detection film and a water absorption pad. The antigen pad absorbs pseudorabies virus detection antigens. The gold labeling pad absorbs colloidal gold labeled anti-pseudorabies virus gE and gB monoclonal antibodies mAb1 and mAb2. Virulent detection line T1''|'', vaccine virus detection line T2''|'' and quality control line C''|'' prints are set on the detection film. The virulent detection line T1 is an anti-pseudorabies virus gE protein monoclonal antibody mAb3 print. The vaccine virus detection line T2 is an anti-pseudorabies virus gB protein monoclonal antibody mAb4 print. The quality control line C is a rabbit anti-mouse IgG antibody pAb1 or staphylococcus aureus SPA print ''|''. According to the test paper, rapid detection and real-time detection of pseudorabies virus infection and immune antibodies are realized, infection and immune detection andimmune evaluation of a vaccine immune effect are realized, operation is simple, and the test paper is easy to popularize and apply in a wide range.

Description

technical field [0001] The invention relates to a detection device for infection and immune antibodies of livestock epidemics, in particular to a test paper for blocking detection of pseudorabies antibodies. Background technique [0002] Pseudorabies (PR) is a zoonotic infectious disease caused by pseudorabies virus (PRV), which can infect cattle, pigs, dogs, sheep, foxes and cats. Pigs are the source of infection and natural storage host of pseudorabies. Pigs infected with PRV show clinical characteristics such as fever, neurological symptoms, high mortality in piglets, respiratory symptoms in large and medium-sized pigs, and abortion in sows. In 2010, pseudorabies was one of the most ideal diseases for large-scale pig farms in my country to carry out disease control, and many pig farms had reached the state of immunity and no infection. However, in 2011, some Bartha-K61 vaccine immunized pig farms had mutated pseudorabies virus, indicating that the traditional gE gene del...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 张改平杨继飞王栋李艳华郭军庆李青梅刘亚伟张晓晨
Owner 河南百奥生物工程有限公司
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