CN1101404C - Anti Bacillus pyocyaneu Flugge vitelline immunoglobulin products and use thereof - Google Patents
Anti Bacillus pyocyaneu Flugge vitelline immunoglobulin products and use thereof Download PDFInfo
- Publication number
- CN1101404C CN1101404C CN98116441A CN98116441A CN1101404C CN 1101404 C CN1101404 C CN 1101404C CN 98116441 A CN98116441 A CN 98116441A CN 98116441 A CN98116441 A CN 98116441A CN 1101404 C CN1101404 C CN 1101404C
- Authority
- CN
- China
- Prior art keywords
- pseudomonas aeruginosa
- igy
- product
- item
- healthcare products
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Abstract
The present invention relates to an anti-pseudomonas aeruginosa vitelline immunoglobulin product. Eggs of poultry immunized by pseudomonas aeruginosa antigens are used for obtaining the product through extraction and purification. The product of the present invention can be made into health products and biological preparations and can be used for preventing and curing primary diseases and secondary infections caused by pseudomonas aeruginosa. The product has favorable activity and specificity.
Description
The present invention relates to a kind of antibody product and application with Yolk immunoglobulin product and application thereof, particularly a kind of anti Bacillus pyocyaneu Flugge of antibody activity.
Pseudomonas aeruginosa belongs to Rhodopseudomonas, can cause the infection at each position of whole body, and secondary infection is particularly common, is a kind of mushroom bigger to human health damage.When human body caused the immunity system function to descend because of a variety of causes, Pseudomonas aeruginosa was easily invaded human body, causes multiple disease or adds the original illness of heavy patient.Though current have some microbiotic that it is had certain effect, this bacterium easily produces resistance and makes its inefficacy.
The object of the present invention is to provide a kind of product with anti Bacillus pyocyaneu Flugge vitelline immunoglobulin (anti--PA IgY) of antibody activity, and use their preparation healthcare products and biotechnological formulations, be used to prevent and treat Pseudomonas aeruginosa protopathy and secondary infection and be used for Pseudomonas aeruginosa and detect diagnostic reagent.
The object of the present invention is achieved like this:
Product with anti Bacillus pyocyaneu Flugge vitelline immunoglobulin (anti--PA IgY) of antibody activity.This product can obtain from following steps: preparation Pseudomonas aeruginosa antigen, and with Pseudomonas aeruginosa antigen immune married woman's parent's home fowl, collect it then and produce the fowl ovum, from the fowl ovum, extract Yolk immunoglobulin again.
Optional hen when selecting married woman's parent's home fowl kind, the hen nursing is convenient and feeding cost is lower.
Can adopt following step for effectively preparing Pseudomonas aeruginosa antigen: from patient's focus, separate obtaining Pseudomonas aeruginosa earlier, then earlier it is inoculated in the common nutrient broth medium tubule, be inoculated in respectively again in the soybean enzymolysis solution substratum and cultivate, culture temperature is 37 ℃, collects nutrient solution and promptly gets antigen.
From yolk, extract Yolk immunoglobulin, adopt water dilution method more superior.This method does not adopt organic digestion agent, and cost is low, is a kind of free of contamination extracting method.The steps include: from birds, beasts and eggs, to collect yolk, with distilled water diluting, standing over night, the centrifuging and taking supernatant liquor concentrates, and lyophilize obtains Yolk immunoglobulin.
Provided by the present invention resisting-PA IgY product is laid eggs poultry by the Pseudomonas aeruginosa antigen immune and is got, has very strong specificity, protopathy and secondary disease that the control Pseudomonas aeruginosa is caused have good effect, and the immunoglobulin (Ig) based article can strengthen the human immunologic function, compare therewith, Pseudomonas aeruginosa easily produces resistance and makes microbiotic lose efficacy.The present invention adopts poultry to produce the fowl ovum to be carrier, and is safe and nontoxic.Poultry particularly hen is easily raised and management, is convenient to industrialization.
Describe the present invention in detail below in conjunction with embodiment.
Embodiment 1: the product with anti Bacillus pyocyaneu Flugge vitelline immunoglobulin (anti--PA IgY) of antibody activity, can obtain from following steps: preparation Pseudomonas aeruginosa antigen, with Pseudomonas aeruginosa antigen immune married woman's parent's home fowl, collect it then and produce the fowl ovum, from the fowl ovum, extract Yolk immunoglobulin again.
Embodiment 2: with Pseudomonas aeruginosa antigen immune hen, all the other steps adopt the method for embodiment 1.
Embodiment 3: separate obtaining Pseudomonas aeruginosa from patient's focus, earlier it is inoculated in the common nutrient broth medium tubule then, be inoculated in respectively in the soybean enzymolysis solution substratum again and cultivate, culture temperature is 37 ℃, collects nutrient solution and promptly gets Pseudomonas aeruginosa antigen.
Embodiment 4: from the birds, beasts and eggs that poultry produced behind the Pseudomonas aeruginosa antigen immune, collect yolk, with distilled water diluting, standing over night, the centrifuging and taking supernatant liquor concentrates, lyophilize, obtain Yolk immunoglobulin (anti--PAIgY).
Embodiment 5: gets any anti--PA IgY product that embodiment 1-4 provides,, carries out indirect hemagglutination test respectively its dilution, single, double to agar diffusion test:
Indirect hemagglutination test: will resist-PA IgY diluent coagulates in the plate aperture between adding, and adds one in 0.5% Pseudomonas aeruginosa sensitization tannic acid blood cell again, puts plate then in the blood clotting micro oscillator, jolting five minutes, 37 ℃, 30 minutes.As seen be the bead-like precipitation in the hole, i.e. the strong positive result.
Unidirectional agar diffusion: get anti--PA IgY diluent and inject incubator, again to wherein adding 2% import agar solution 9ml, abundant mixing, the punching of cooling back adds and boils inactivation Pseudomonas aeruginosa liquid, observations after 37 ℃, 24 hours.The result all forms the precipitation circle around visible each hole.
Two-way agar diffusion: on 1.5% import agar plate, beat quincunx hole.Interstitial hole drips anti--PA IgY diluent, and what peripheral hole added different concns respectively boils inactivation Pseudomonas aeruginosa liquid, and the intestinal bacteria inactivated bacterial liquid, streptococcus aureus inactivated bacterial liquid, observations after 37 ℃, 48 hours.The result is as seen anti--PA IgY hole and Pseudomonas aeruginosa hole between the clear precipitation line of formation; And the intestinal bacteria hole, streptococcus aureus hole and anti--PA IgY do not have precipitation line between the hole.
Above result has shown that anti--PA IgY has effect of specificity anti Bacillus pyocyaneu Flugge and higher antibody activity.Simultaneously show that also anti--PA IgY can be used as the detection diagnostic reagent of Pseudomonas aeruginosa.Embodiment 6: because anti--PA IgY is to specific effect of Pseudomonas aeruginosa and higher antibody activity, it can be used for preventing and treating Pseudomonas aeruginosa protopathy and secondary disease, any anti--PA IgY product that embodiment 1-4 is provided is used to prevent and treat the respiratory tract infection that Pseudomonas aeruginosa causes, pneumonia, otitis media, keratitis, urethritis, endocarditis, gastro-enteritis, the Pseudomonas aeruginosa secondary infection of septicemia and burn and wound.Embodiment 7: by anti--healthcare products and biotechnological formulation of being used to prevent and treat Pseudomonas aeruginosa protopathy and secondary infection that PA IgY product is formed.Embodiment 8: the product that contains anti--PA IgY is used to prevent and treat the healthcare products and the biotechnological formulation of Pseudomonas aeruginosa protopathy and secondary infection.Embodiment 9: healthcare products and the biotechnological formulation of using anti--PA IgY product preparation control Pseudomonas aeruginosa protopathy and secondary infection.Embodiment 10: will resist-PA IgY is used to prevent and treat the intestinal bacterial translocation due to the portal hypertension, strengthens the human immunologic function.
Patients with cirrhosis is easy to take place main spontaneous microbemia and the peritonitis that is caused by intestinal bacteria, and portal hypertension causes intestines wall gram blood, and oedema and intestinal bacterium hyper-proliferative impel the interior bacterium of enteric cavity to send out to regional lymph nodes or systemic circulation, i.e. bacterial translocation.Experiment shows that intestinal bacterial translocation is an important early process of liver cirrhosis spontaneous peritonitis.
With the SD rat is research object, is divided into following three groups at random:
Portal hypertension control group: anti--PA IgY2ml is joined in perfusion in the experiment glandular stomach, once a day, and continuous three days; Form preceding 24 hours of portal hypertension model and pour into Pseudomonas aeruginosa living bacterial liquid 10 through stomach
8/ kg; Finish model and pour into anti--PA IgY2ml after 8 hours again through stomach; 1% Sodital anesthesia after 48 hours, aseptic technique is got required sample and is equipped with inspection.
Portal hypertension group: do not do special processing before the experiment, pour into Pseudomonas aeruginosa living bacterial liquid 10 through stomach in forming preceding 24 hours of model
8/ kg; Finish model after 48 hours, the anesthesia of 1% Sodital, aseptic technique are got required sample and are equipped with inspection.
Control group: pour into Pseudomonas aeruginosa living bacterial liquid 10 through stomach
8/ kg, after 24 hours, separation gate vein but non-constriction, collection of specimens is with portal vein high pressure control group.
More than three groups of SD rats got mesenteric lymph nodes, portal vein and painstaking effort in two days respectively at postoperative and make microbial culture, to the intestinal bacteria quantitative culture, first mirror is observed liver, ileum pathological change down.The result shows: portal hypertension group mesenteric lymph nodes and portal vein blood bacterium recall rate obviously raise, but the peripheral blood cultivation is negative; There were significant differences for a remarkable rising of portal vein blood level of endotoxin and a door body circulation intracellular toxin, and the intestinal contents level of endotoxin obviously raises; The entero-bacte amount obviously increases; Visual inspection, liver surface are sliding earlier, and color and luster is ruddy, and matter is soft, the ileum expansion, unstriated muscle is loose, full of liquid and gas in the chamber, and intestines wall extravasated blood is also shown in facial intestinal mucosa erosion, and is hemorrhage, under elder generation's mirror, the slight edema of liver, the kitchen range lymphocytic infiltration, liver lobule structure and liver cell form are complete substantially; The ileum pathological change is remarkable, and the intestinal epithelial cells necrosis comes off, a small amount of inflammatory cell infiltration, the slight extravasated blood of mucous membrane is hemorrhage.Its mesenteric lymph nodes is organized in the portal hypertension control, portal vein blood bacterium recall rate obviously reduces, and blood and intestinal contents level of endotoxin significantly descend, and the intestinal bacteria hyper-proliferative obviously is subjected to press down, and liver, ileum pathological change significantly alleviate.
By above test as seen, portal hypertension is impelled intestinal bacteria and endotoxin translocation, the intestinal mucosal barrier function is impaired due to the portal hypertension, the intestinal microecology equilibrated destroys and intestinal endotoxemia is the important precipitating factor of bacterial translocation, anti--PA IgY can alleviate intestinal mucosal injury, safeguard the intestinal microecology balance and reduce level of endotoxin, have the effect of control intestinal bacteria, endotoxin translocation.Embodiment 11: adopt burn rat Pseudomonas aeruginosa enterogenic infection animal model, with give the oral specificity of burn rat anti--method of PA IgY, investigate its influence to serious burn rat enterogenic infection.
The healthy SD rat is divided into the normal group that contrasts usefulness at random, the burn group of contrast usefulness and anti--PA IgY prevention group:
After the rat fasting 8 hours, inject 1.5%NaHCO in the stomach tube
31ml treats to inject 10 after one minute
8The bacteria suspension of the Pseudomonas aeruginosa of CFU/ml (1ml/100g), ad lib.Give behind the bacterium 8 hours, ketamine intramuscular injection (30mg/kg) anesthesia down, back 10% Na
2The S depilation.Normal group is not done any processing.Burn group and IgY prevention group are distinguished 20 seconds with 100 ℃ of piece of stainless steel contact depilations, cause 25% TBSA third degree burn.Intraperitoneal injection of saline (3ml/100g) antishock immediately after the wound, the surface of a wound is coated with the Sulfadiazine Silver creme immediately, and per 24 hours are once.Behind the IgY group burn immediately tube feed anti--PA IgY2ml, per 12 hours repeat once, up to sacrifice of animal.Burn group is capacity physiological saline such as tube feed then.Ad lib after 24 hours is freely drunk water in the single cage management of animal.Three treated animals all in causing injury back 24 hours and work in 48 hours is killed, are dissected, and carry out every test:
Test-results shows that use the burn back, and anti--PA IgY prevention significantly descends portal vein blood endotoxin content.Burnt back 24 hours, put all mutually and can reflect two the time in 48 hours: burn group portal vein blood endotoxin content is significantly higher than normal group (P＜0.01); IgY group portal vein blood endotoxin content significantly is lower than burn group (P＜0.01); IgY group portal vein blood endotoxin content and normal group endotoxin content indifference (P＞0.05).In addition, mesenteric lymph nodes microbial culture normal group is all negative, burn group was burnt back 24 hours, mesenteric lymph nodes bacterial translocation incidence was respectively 85.7% in 48 hours, 80%, compare 0%, 20% with the IgY group, P＜0.01 burn group and IgY group burn back hemoculture result analyze P＜0.01 by statistics.
Combining form is learned and is observed, and shows the severity and the intestinal mucosa pathological change basically identical of enterogenic infection.Positive bacterial culture can cause Sepsis after showing serious burn in the blood.Thus, evidence: on this animal model, use anti--PA IgY after plasma endotoxin obviously descend, intestinal mucosa lymphoglandula positive rate of bacteria reduces, the enteron aisle physical barriers is protected, thereby has reduced the generation of enterogenic infection.Anti--PA IgY is easy to use, can add patients wet suit usefulness, and this is that other enterogenic infection means of prevention is incomparable.
Claims (10)
1, a kind of anti Bacillus pyocyaneu Flugge vitelline immunoglobulin (anti--PA IgY) product with antibody activity, this product can obtain from following steps, at first from patient's focus, separate and obtain Pseudomonas aeruginosa, then earlier it is inoculated in the common nutrient broth medium tubule, cultivate by planting in soybean enzymolysis solution substratum respectively again, culture temperature is 37 ℃, collect nutrient solution and obtain Pseudomonas aeruginosa antigen, with Pseudomonas aeruginosa antigen immune married woman's parent's home fowl, collect it then and produce the fowl ovum, from the fowl ovum, collect yolk again, with distilled water diluting, standing over night; The centrifuging and taking supernatant liquor concentrates, and lyophilize obtains Yolk immunoglobulin.
2, according to claim 1 resisting-PA IgY product, it is characterized in that: the poultry by immunity is a hen.
3, by in the claim 1-2 item any one described anti--PA IgY product being used to of forming prevent and treat the healthcare products and the biotechnological formulation of the protopathy that Pseudomonas aeruginosa causes.
4, contain in the claim 1-2 item any one described anti--PA IgY product, be used to prevent and treat the healthcare products and the biotechnological formulation of the protopathy that Pseudomonas aeruginosa causes.
5, application rights requires the healthcare products and the biotechnological formulation of any one described resisting in the 1-2 item-protopathy that PA IgY product preparation control Pseudomonas aeruginosa causes.
6, by any one healthcare products and biotechnological formulation of being used to prevent and treat the Pseudomonas aeruginosa secondary infection that described resisting-PA IgY product is formed in the claim 1-2 item.
7, contain in the claim 1-2 item any one described anti--PA IgY product is used to prevent and treat the healthcare products and the biotechnological formulation of Pseudomonas aeruginosa secondary infection.
8, application rights requires the healthcare products and the biotechnological formulation of any one described resisting in the 1-2 item-PA IgY preparation control Pseudomonas aeruginosa secondary infection.
9, Pseudomonas aeruginosa protopathy and the secondary disease according to any one described resisting-PA IgY healthcare products and biotechnological formulation control in the claim 3-8 item is following disease: respiratory tract infection, pneumonia, otitis media, keratitis, urethritis, endocarditis, gastro-enteritis, septicemia, and burn, wound generation charrin's disease.
10, be used as the detection diagnostic reagent of Pseudomonas aeruginosa by any one described resisting-PA IgY product in the claim 1-2 item.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN98116441A CN1101404C (en) | 1998-07-29 | 1998-07-29 | Anti Bacillus pyocyaneu Flugge vitelline immunoglobulin products and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN98116441A CN1101404C (en) | 1998-07-29 | 1998-07-29 | Anti Bacillus pyocyaneu Flugge vitelline immunoglobulin products and use thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1208732A CN1208732A (en) | 1999-02-24 |
| CN1101404C true CN1101404C (en) | 2003-02-12 |
Family
ID=5225073
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN98116441A Expired - Fee Related CN1101404C (en) | 1998-07-29 | 1998-07-29 | Anti Bacillus pyocyaneu Flugge vitelline immunoglobulin products and use thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1101404C (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100360566C (en) * | 1999-09-08 | 2008-01-09 | 中德联合研究院 | Pyloric bacillary spirillum resisting yolk immunoglobulin product and its application |
| CA2925897A1 (en) * | 2013-09-30 | 2015-04-02 | Daiichi Sankyo Company, Limited | Anti-lps o11 antibody |
| CN108992669B (en) * | 2018-08-27 | 2022-03-22 | 广州汇高生物科技有限公司 | Composite yolk antibody composition for preventing and treating respiratory tract infection, aerosol inhalation solution, preparation process and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1060109A (en) * | 1990-09-25 | 1992-04-08 | 铁汉 | High efficient sterilizing toilet soap |
| CN1090602A (en) * | 1993-02-06 | 1994-08-10 | 牟希亚 | Pseudomonos aeruginosa MSHA fimbria strain and foundation thereof |
| US5367054A (en) * | 1993-04-12 | 1994-11-22 | Stolle Research & Development Corp. | Large-scale purification of egg immunoglobulin |
-
1998
- 1998-07-29 CN CN98116441A patent/CN1101404C/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1060109A (en) * | 1990-09-25 | 1992-04-08 | 铁汉 | High efficient sterilizing toilet soap |
| CN1090602A (en) * | 1993-02-06 | 1994-08-10 | 牟希亚 | Pseudomonos aeruginosa MSHA fimbria strain and foundation thereof |
| US5367054A (en) * | 1993-04-12 | 1994-11-22 | Stolle Research & Development Corp. | Large-scale purification of egg immunoglobulin |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1208732A (en) | 1999-02-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Williams et al. | The role of the O and K antigens in determining the resistance of Klebsiella aerogenes to serum killing and phagocytosis | |
| Ali et al. | Pathological and immunological study on infection with Escherichia coli in ale balb/c mice | |
| CN113583972B (en) | Escherichia coli bacteriophage capable of reducing antibiotic resistance and application thereof | |
| CN106434411B (en) | A kind of giant panda source lactobacillus plantarum and application | |
| US6632432B1 (en) | Directed human immune globulin for the prevention and treatment of staphylococcal infections | |
| CN101104637A (en) | Cecropin and its generation bacterial strain and use | |
| CN1101404C (en) | Anti Bacillus pyocyaneu Flugge vitelline immunoglobulin products and use thereof | |
| Kothary et al. | Purification and characterization of a vulnificolysin-like cytolysin produced by Vibrio tubiashii | |
| Sack et al. | Experimental Canine Cholera: II. Production by Cell-free Culture Filtrates of Vibrio cholerae | |
| JPH08504167A (en) | Directed human immunoglobulins for prevention and treatment of Staphylococcus infection | |
| CN105754886B (en) | One plant of nontoxic bacillus cereus strain pBC-1 and its application | |
| Dash et al. | Biochemical and serological characterization of Flavobacterium columnare from freshwater fishes of Eastern India | |
| Ariyanti et al. | Contamination of Escherichia coli O157: H7 in milk and its dairy products in Depok, Cianjur, Sukabumi and Bandung | |
| Afshar et al. | A study on Haemophilus influenzae type b growth rate and capsule production in different media | |
| CN101570574B (en) | Anti-I type Shiga toxin IgY antibody as well as preparation method and use thereof | |
| Goldwater et al. | Hemolytic uremic syndrome due to shiga-like toxin producing Escherichia coli 048: H21 in South Australia. | |
| CN103800901B (en) | Streptococcus and the mammitis of cow vaccine that its deactivation is obtained | |
| CN1059128C (en) | Milk powder containing specific immunity globulin and its prodn. method | |
| CN108018228A (en) | One plant of Escherichia coli O 157:H7 low virulent strains and its application | |
| Aucher et al. | Meningitis due to enterotoxigenic Bacteroides fragilis | |
| CN107064504A (en) | A kind of method of lysozyme in detection egg | |
| Megha et al. | Generation and Characterization of specific Chicken Egg Yolk Antibodies (IgY) against microbial bio-terroristic Agent (Vibrio cholerae) | |
| CN114042153B (en) | Milk cow mastitis multi-linked inactivated vaccine and preparation method and application thereof | |
| CN113546100B (en) | Application of pachyrhizus extract in preparation of antibacterial product | |
| CN105884890A (en) | Citrobacter freundii resisting yolk antibody, preparation method therefor and application of Citrobacter freundii resisting yolk antibody |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |