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Application of lipopolysaccharide extracted from Brucella melis vaccine strain m5 in the preparation of products for diagnosing human brucellosis

A technology for brucella and brucellosis, which is applied in the field of antigen extraction from live brucella vaccines, can solve the problems of lack of O antigen components, weak virulence and smooth type

Active Publication Date: 2021-08-17
新疆禹孚生物技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The rough LPS of Clothella lacks O antigen components, and its virulence is weaker than that of the smooth type

Method used

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  • Application of lipopolysaccharide extracted from Brucella melis vaccine strain m5 in the preparation of products for diagnosing human brucellosis
  • Application of lipopolysaccharide extracted from Brucella melis vaccine strain m5 in the preparation of products for diagnosing human brucellosis
  • Application of lipopolysaccharide extracted from Brucella melis vaccine strain m5 in the preparation of products for diagnosing human brucellosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] The LPS antigen of embodiment 1, Brucella melis vaccine strain M5 is extracted

[0077] Tested strains: Brucella melis vaccine strain M5, Clothella pig strain S2, Clothella bovine strain S19, Shigella flexneri (Shigella flexneri was isolated from inpatients in the First Affiliated Hospital of Xinjiang Medical University , identified as Shigella flexneri, the patient gave informed consent), Staphylococcus aureus (Staphylococcus aureus was isolated from an inpatient in the First Affiliated Hospital of Xinjiang Medical University, identified as Staphylococcus aureus, the patient gave informed consent) .

[0078] 1. Strain cultivation and harmless treatment

[0079] Before the start of the experiment, all experimental operators should take protective measures, wear disposable gloves, disposable surgical protective clothing, disposable protective masks, disposable shoe covers, etc., and operate in a biological safety cabinet. The experimental operation steps are as follows...

Embodiment 2

[0094] Example 2, Screening, confirmation and application of the LPS antigen extracted from Brucella melis vaccine strain M5 as human brucellosis diagnostic antigen

[0095] Using the LPS extracted from Brucella melis vaccine strain M5, Clothella pig strain S2, Clothella bovine strain S19, and Shigella flexneri respectively in Example 1, the peptidoglycan extracted from Staphylococcus aureus Sugar, as well as LPS finished products purchased from Hangzhou Yiminuo Biotechnology Co., Ltd. combined with immuno-infiltration gold standard detection technology to compare the reactivity of LPS or peptidoglycan from different sources as diagnostic antigens. Specific steps are as follows:

[0096] 1. Add 0.5 μL of antigens from different sources and different concentrations to the nitrocellulose membrane (NC membrane, pore size 0.45 μm, GE Company, Cat. No. 10600002) in the sample well, and wait for the membrane to be absorbed. The injection position of each antigen is as follows: fig...

Embodiment 3

[0112] Embodiment 3, the preparation of colloidal gold test strip and its sensitivity and specificity detection

[0113] 1. Preparation of colloidal gold test strips

[0114] 1. Preparation of detection pad containing detection line T and quality control line C

[0115] 1) Preparation of coated antigen

[0116] The LPS extracted from the Brucella melis vaccine strain M5 in Example 1 was prepared into an LPS antigen solution with a concentration of 0.01 mg / mL (the solvent was Tris buffer).

[0117] 2) Preparation of detection pads containing detection line T and quality control line C

[0118] On the nitrocellulose membrane (Millipore Hi-Flow Puls HFB13502), the test line (T line) and the quality control line (C line) were respectively coated: the T line was coated with the Brucella melis vaccine strain M5 to extract the LPS antigen solution ( Use pH 8.0, Tris-HCl buffer diluted to 0.01mg / mL dashed line); C line coated with goat anti-human IgG antibody (purchased from Sigma)...

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Abstract

The invention discloses the application of the lipopolysaccharide extracted from the brucella melis vaccine strain M5 as an antigen for diagnosing human brucellosis. The present invention finds that the reactivity of the LPS antigen extracted from the Brucella melis vaccine strain M5 as a diagnostic antigen is significantly better than the Brucella bovine strain S19 strain and the Brucella pig strain S2 strain commonly used in the current brucellosis antibody rapid detection kits on the market The extracted LPS antigen shows that the Brucella melis vaccine strain M5 can be used as the preferred source strain of LPS antigen for the new gold standard diagnosis of brucellosis, and the extracted LPS has good diagnostic value for human brucellosis The selection of antigens for infection diagnosis provides new avenues. The present invention also finds that the LPS antigen extracted from the detoxified Brucella melis vaccine strain M5 does not affect its antigenicity.

Description

technical field [0001] The invention relates to the technical field of application of antigens extracted from live Brucella vaccines, in particular to the application of lipopolysaccharide extracted from Brucella melis vaccine strain M5 as a diagnostic antigen for human brucellosis. Background technique [0002] Brucellosis (brucellosis, referred to as brucellosis) is a serious zoonotic infectious disease caused by Brucella (brucella, referred to as brucellosis), which has caused great harm to human health and also caused great harm to animal husbandry. serious economic loss. World statistics show that the annual economic loss caused by brucellosis is nearly 3 billion US dollars. Human wave fever and low-grade fever caused by it, heart, bone and nervous system lesions caused by chronic infection, as well as diseases such as abortion and orchitis in ruminants are still incurable. In addition, Brucella has been used as a biological weapon. It is particularly prevalent in ou...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/00G01N33/558G01N33/569G01N33/58
CPCC08B37/0003G01N33/558G01N33/56911G01N33/587G01N2333/23G01N2469/10
Inventor 张文宝郭刚李军游锡火贾斌齐文静吴熙然郭宝平
Owner 新疆禹孚生物技术股份有限公司
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