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Helicobacter pylori virulence hypotype mixed representative strain group and immunized cow milk prepared from same

A technology for Helicobacter pylori and Helicobacter pylori, applied in the biological field, can solve problems such as easy infection and inability to cover Helicobacter pylori, and achieve the effects of low cost, no drug resistance, and good patient compliance

Active Publication Date: 2018-10-16
FOURTH HOSPITAL OF HEBEI MEDICAL UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among these outer membrane proteins, the receptors that bind to them have been confirmed, and the adhesion receptors of many outer membrane proteins are not yet clear, so the vaccines currently prepared by recombinant methods cannot cover most infected people. Helicobacter pylori in the stomach. Previous studies have shown that people infected with different blood types have different infection states due to different receptors expressed. For example, blood type O is more likely to express Le b Antigen, O blood is more likely to be infected with Helicobacter pylori, so it is necessary to group the Helicobacter pylori in the stomach of the infected person according to the blood type

Method used

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  • Helicobacter pylori virulence hypotype mixed representative strain group and immunized cow milk prepared from same
  • Helicobacter pylori virulence hypotype mixed representative strain group and immunized cow milk prepared from same
  • Helicobacter pylori virulence hypotype mixed representative strain group and immunized cow milk prepared from same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 Isolation, cultivation and identification of gastric Helicobacter pylori

[0064] The isolated population came from a total of 498 cases of Helicobacter pylori infection in Shijiazhuang area. Bite the gastric mucosal tissue of Helicobacter pylori infected patients through gastroscope, and then put the gastric mucosal tissue into a glass grinder containing 0.5ml of Brooke's broth and grind it into a slurry, draw 100 microliters and spread it on the Columbia selection medium Plate (containing 8% defibrillated sheep blood, 5mg / L TMP, 5mg / L polymyxin B, 5mg / L amphotericin B, 10mg / L vancomycin), and then place the plate in a three-gas culture Cultivated in the box (10% CO 2 , 85% N 2 , 5% O 2 ), cultured at 37°C for 3-7 days, and carried out Gram staining microscopic examination, urease, catalase, and oxidase experiments on the suspicious colonies cultured. The result is Gram-negative, S-type or arc-type, and the positive three-enzyme test can be initially iden...

Embodiment 2

[0065] Example 2 Gene PCR method of Helicobacter pylori

[0066] (1) Bacterial DNA extraction method

[0067] 1. Scrape a fresh bacterial lawn (soybean size) from the Helicobacter pylori medium and place it on the wall of a 1.5ml EP tube. Add 0.9% normal saline, shake and mix well, centrifuge at 10000rpm for 2 min, discard the supernatant, and repeat this 3 times;

[0068] 2. Add 180ul ATL (to lyse the bacteria and dissolve the precipitate);

[0069] 3. Add 20ul of proteinase K, vortex to mix, put in a 56-degree metal bath at 300rpm for 1-3 hours, and centrifuge briefly;

[0070] 4. Add 200ul AL, shake and mix, 70 degree metal bath for 10mins, and centrifuge instantly;

[0071] 5. Add 200ul of absolute ethanol, gently mix up and down until clarified for 15 seconds, and centrifuge briefly;

[0072] 6. Add 500ul AW1, centrifuge at 8000rpm for 1min, discard the tube and keep the column, and replace with a new collection tube;

[0073] 7. Add 500ul AW2, centrifuge at 14000rpm...

Embodiment 3

[0085] Example 3 Preparation of Dairy Cow Vaccine by Helicobacter pylori Virulence Subtype Mixed Representative Strain Group

[0086] (A) Helicobacter pylori infected patients were divided into A, B, and O groups according to the ABO blood group system, and Helicobacter pylori was isolated from the stomachs of patients infected with different blood types of Helicobacter pylori;

[0087] (B) PCR and sequencing of cagA, cagE, vacA, iceA, babA2 of Helicobacter pylori isolated from each blood type

[0088] (C) After sequencing cagA, cagE, vacA, iceA, babA2, according to blood type and gene combination: genotype cagA was screened from group A Helicobacter pylori + / cagE2 / s1a / m2 / i1 / iceA1, genotype cagA was screened from Helicobacter pylori group B + / cagE2 / s1a / m1b / i1 / iceA1, genotype cagA was screened from Helicobacter pylori group O + / cagE2 / s1a / m2 / babA2 / i1 / iceA1 three representative strains;

[0089] (D) Three popular representative strains of virulent subtypes were collected by...

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Abstract

The invention provides a helicobacter pylori virulence hypotype mixed representative strain group, a dairy cow vaccine prepared by using the representative strain group, immunized cow milk prepared byimmunized cows with the dairy cow vaccine, and biological products prepared by using polyclonal IgG antibodies in the immunized cow milk, so that the requirements of clinical specific treatment of helicobacter pylori infection are met, at the same time, the problem of bacterial drug resistance is solved. The helicobacter pylori virulence hypotype mixed representative strain group and the immunized cow milk prepared from the same have the advantages of low cost, good patient compliance, long-term drinking of the milk, and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a Helicobacter pylori virulence subtype mixed representative strain group, a cow vaccine prepared by using the Helicobacter pylori virulence subtype mixed representative strain group, and an immune vaccine obtained after immunizing cows with the cow vaccine. Cow's milk and a technical method for preparing biological products by using polyclonal IgG antibodies in immunized cow's milk and its application method. Background technique [0002] Helicobacter pylori, also known as Helicobacter pylori, is a pathogenic bacterium that has co-evolved with humans, and infection usually occurs in infancy. It is estimated that more than half of the world's population is infected with Helicobacter pylori, but 80% of infected people present as chronic gastritis without obvious clinical symptoms; about 15-20% of infected people develop ulcers; 1-5% of infected people The inflammatory resp...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12Q1/6869C12Q1/04C07K16/42A61K39/02A61P31/04A61K39/395A01K67/02A23C7/04A23C3/04A61K35/20A61K33/00C12R1/01
CPCA01K67/02A23C3/04A23C7/04A61K33/00A61K35/20A61K39/0208A61K39/39566A61K2039/552A61P31/04C07K16/42C12Q1/6869C12N1/205C12R2001/01A61K2300/00C12Q2531/113
Inventor 单保恩赵川史中立胡代伦赵连梅刘维华徐保红殷长甫王苋
Owner FOURTH HOSPITAL OF HEBEI MEDICAL UNIV