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Detection method and detection kit of antigen-specific IgG (immunoglobulin G) antibody of staphylococcus aureus SpA5 mutant

A kit and specific technology, applied in the field of antigen-specific IgG antibody detection methods and detection kits, can solve problems such as difficulty in detecting SpA5-specific IgG, and achieve improved specificity, simple operation and good repeatability Effect

Active Publication Date: 2014-03-19
CHENGDU OLYMVAX BIOPHARM +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the SpA5 mutant can still partially bind to the Fc region of mammalian IgG antibodies, which makes it difficult to detect SpA5-specific IgG.

Method used

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  • Detection method and detection kit of antigen-specific IgG (immunoglobulin G) antibody of staphylococcus aureus SpA5 mutant
  • Detection method and detection kit of antigen-specific IgG (immunoglobulin G) antibody of staphylococcus aureus SpA5 mutant
  • Detection method and detection kit of antigen-specific IgG (immunoglobulin G) antibody of staphylococcus aureus SpA5 mutant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: Immunization of animals and the acquisition of immune serum

[0038] 1) Grouping of animals: According to the principle of randomization, the animals were divided into vaccine immunization group and control group. The grouping situation is shown in Table 1.

[0039] Table 1: Grouping of experimental animals for immunogenicity studies

[0040]

[0041]2) Immunization of experimental animals: the vaccine group was injected intramuscularly with SpA5 protein, one injection (600 μL) per animal; the control group was immunized with the same volume of normal saline, and the immunization procedure was three times on day 0, day 14 and day 21.

[0042] 3) Blood collection: Blood was collected 14 days after the end of the last immunization. Blood was collected from mice by enucleation of the eyeballs, from rats by tail veins, and by rabbits from ear veins. After the blood was incubated at 4°C for 2 hours, it was centrifuged at 8000rpm for 10min, and the serum was...

Embodiment 2

[0043] Example 2: Pepsin Digestion of Antibodies in Serum

[0044] Take 20 μl of the serum obtained in Example 1, add 180 μl of enzyme digestion solution, mix well, and digest in a water bath at 37°C for 6 hours, shake and mix for 5 minutes every 1 hour.

Embodiment 3

[0045] Embodiment 3: the detection of anti-SpA5 specific antibody

[0046] experimental method:

[0047] 1) Coating: Dilute the purified SpA5 protein to 2 μg / mL with coating solution. Coat the ELISA plate with 100 μL / well, shake well and spread evenly, and place in a refrigerator at 4°C overnight or at 37°C for 2 hours.

[0048] 2) Sealing: wash the plate 4 times with washing solution, add 300 μL of solution each time, shake for 30 s, and absorb the solution for 2.5 s. Block the enzyme strips with 200 μL / well of blocking solution, and place in a refrigerator at 4°C overnight or at 37°C for 2 hours.

[0049] 3) Add the primary antibody: Wash the plate 4 times with washing solution, add 300 μL each time, shake for 30 s, and absorb the solution for 2.5 s. Dilute the enzyme-digested serum by 200 times to 1:2000, then dilute 7 gradients by 2 times with antibody diluent, shake well. The diluted sample was added to the enzyme-linked plate at 100 μL / well, and incubated in a 37°C i...

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Abstract

The invention belongs to the technical field of biology, and relates to a detection method and a detection kit of an antigen-specific IgG (immunoglobulin G) antibody of a staphylococcus aureus (SA) SpA5 mutant. By adopting the method disclosed by the invention, the specificity of the method is improved, and the method is simple and fast in operation, and good in repeatability.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a Staphylococcus aureus (SA) SpA5 mutant, and more specifically, to a detection method and a detection kit for an antigen-specific IgG antibody of the mutant. Background technique [0002] Staphylococcus aureus (Staphylococcus aureus, SA), hereinafter referred to as Staphylococcus aureus, has another name of "flesh-loving bacteria". As a representative of Gram-positive bacteria, it is an important pathogenic bacteria causing nosocomial infection and community infection. Infection is characterized by acute, purulent inflammation. Local infection can cause purulent infection of the skin and soft tissues, which does not heal after a long time; systemic infection can lead to osteomyelitis, septic arthritis, endocarditis, pneumonia, and sepsis And other serious infections and complications, the mortality rate is as high as 20%. At the same time, the exotoxin of Staphylococcus aureus can a...

Claims

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Application Information

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IPC IPC(8): G01N33/569
CPCG01N33/56911G01N33/6803G01N2333/31
Inventor 赵莉群章金勇邹全明曾浩顾江纪永军
Owner CHENGDU OLYMVAX BIOPHARM
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