CN105784984A - Preparation method and application of gram-positive bacterium ghost - Google Patents

Preparation method and application of gram-positive bacterium ghost Download PDF

Info

Publication number
CN105784984A
CN105784984A CN201610116309.6A CN201610116309A CN105784984A CN 105784984 A CN105784984 A CN 105784984A CN 201610116309 A CN201610116309 A CN 201610116309A CN 105784984 A CN105784984 A CN 105784984A
Authority
CN
China
Prior art keywords
gram
positive bacteria
preparation
bacterium shadow
suspension
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.)
Granted
Application number
CN201610116309.6A
Other languages
Chinese (zh)
Other versions
CN105784984B (en
Inventor
鞠兴荣
都立辉
吴学友
袁建
何荣
陈正行
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nanjing University of Finance and Economics
Original Assignee
Nanjing University of Finance and Economics
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Nanjing University of Finance and Economics filed Critical Nanjing University of Finance and Economics
Priority to CN201610116309.6A priority Critical patent/CN105784984B/en
Publication of CN105784984A publication Critical patent/CN105784984A/en
Application granted granted Critical
Publication of CN105784984B publication Critical patent/CN105784984B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56911Bacteria

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Food Science & Technology (AREA)
  • Biochemistry (AREA)
  • Cell Biology (AREA)
  • Biotechnology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

The invention provides a preparation method and application of a gram-positive bacterium ghost, and belongs to the field of microbiology and immunology.The preparation method of the ghost includes the steps that NaOH, SDS and CaCO3 are added into gram-positive bacterium suspension, incubation is carried out at 30-40 DEG C, centrifugation is carried out, precipitate is collected, and after washing, resuspension A is prepared; H2O2 is added into the resuspension A, incubation is carried out at 30-40 DEG C, centrifugation is carried out, precipitate is collected, and after washing, resuspension B is prepared; the resuspension B is incubated at 20-30 DEG C, centrifugation is carried out, precipitate is collected, washing is carried out, and the gram-positive bacterium ghost is obtained.According to the preparation method, the gram-positive bacterium ghost can be rapidly and efficiently prepared with simple chemical reagents, and thus production cost is significantly reduced.As the integrity of a bacterium structure is retained in a Listeria monocytogenes ghost, antibodies for Listeria monocytogenes can be generated after immunization, and thus antibodies capable of efficiently and rapidly detecting Listeria monocytogenes can be obtained through screening.

Description

The preparation method and applications of gram-positive bacteria bacterium shadow
Technical field
The invention belongs to microbiology and field of immunology, be specifically related to the preparation side of gram-positive bacteria bacterium shadow Method.
Background technology
Bacteria simulacrum (Bacterial Ghost) is a kind of empty bacterial body not having cytoplasm and nucleic acid, and it remains The integrality of bacterial structure, remains former somatic surface antigen determinant, screens after therefore may be used for immune animal For the antibody of this bacterium, can efficiently, quickly detect the antibody of this bacterium with exploitation.At present, both at home and abroad In terms of the research of bacteria simulacrum, mostly use cracking system based on bacteriophage Φ X174 Lysis gene E Animal nutrition, and it is directed to gramnegative bacterium, such as: Escherichia coli, Salmonella typhimurium Bacterium, Bacterium enteritidis, haemophilus influenzae, Actinobacillus pleuropneumoniae, helicobacter pylori, pneumonia Klebsiella, haemolysis Pasteurella, pasteurella multocida, comma bacillus, Pseudomonas aeruginosa, stench vacation Monad etc..But, the method is for the preparation of gram-positive bacteria bacterium shadow inapplicable.
Summary of the invention
It is an object of the invention to provide one utilizes simple chemical reagent just can quickly, efficiently prepare leather orchid The method of family name's positive bacteria bacterium shadow.
The preparation method of gram-positive bacteria bacterium shadow, comprises the steps:
(1) in Gram-positive bacteria suspension, NaOH, SDS and CaCO are added3, under the conditions of 30-40 DEG C Hatch, centrifugal, collect precipitation, after washing, make re-suspension liquid A;
(2) in described re-suspension liquid A, H is added2O2, hatch under the conditions of 30-40 DEG C, centrifugal, collect precipitation, Re-suspension liquid B is made after washing;
(3) re-suspension liquid B is hatched under the conditions of 20-30 DEG C, centrifugal, collect precipitation, washing, obtain leather orchid Family name's positive bacteria bacterium shadow.
Preferably in technical scheme, the end of the NaOH added in Gram-positive bacteria suspension in step (1) is dense Degree is the final concentration of 0.1-0.3mmol/L of 20-40mmol/L, the SDS of addition, and every liter of gram-positive bacteria hangs CaCO in liquid3Addition be 9-11mmol;In step (1), in 80-120r/min condition when suspension is hatched Lower shaking, incubation time is 40-60min.
Preferably in technical scheme, the H added in re-suspension liquid A described in step (2)2O2Final mass dense Degree is 0.02-0.05%;In step (2), shake under the conditions of 80-120r/min when suspension is hatched, when hatching Between be 20-40min.
Preferably in technical scheme, in step (3), when described re-suspension liquid B is hatched under the conditions of 30-50r/min Shaking, incubation time is 20-40min.
Preferably in technical scheme, physiological saline is used to wash.
Preferably in technical scheme, in step (2), re-suspension liquid B is the second that described precipitation is resuspended in 55-65% Gained in alcohol solution.
Preferably in technical scheme, described re-suspension liquid A is that described precipitation is resuspended in gained after water.
Preferably in technical scheme, described gram-positive bacteria is Listeria monocytogenes.
Preferably in technical scheme, the OD of described Gram-positive bacteria suspension600It is 0.3~0.5.
The present invention also provides for bacterium shadow prepared by said method in preparation for detecting the anti-of described gram-positive bacteria Application in terms of body.
The present invention utilizes simple chemical reagent just can quickly, efficiently prepare gram-positive bacteria bacterium shadow, significantly Reduce production cost.Listeria monocytogenes (is abbreviated as Listeria monocytogenes, Listeria Monocytogenes) it is the pathogen that a kind of danger is serious, serious infectious diseases common to human beings and animals can be caused, Such as symptoms such as meningitis, septicemia, miscarriage and monocytosis, it the listeriosis caused is lethal Rate is more than 25%.Owing to Listeria monocytogenes bacterium shadow remains the integrality of bacterial structure, especially remain former bacterium Surface antigenic determinant, can produce the antibody for Listeria monocytogenes, thus screen energy after immunity Enough antibody efficient, quickly this bacterium of detection.
Accompanying drawing explanation
The SEM observed result of Fig. 1 Listeria monocytogenes bacterium shadow, in figure A, the object of observation is single increasing Liszt Bacterium living cells, in figure B, the object of observation is Listeria monocytogenes bacterium shadow.
The tem observation result of Fig. 2 Listeria monocytogenes bacterium shadow, in figure A, the object of observation is Listeria monocytogenes Living cells, in figure B, the object of observation is Listeria monocytogenes bacterium shadow.
The antibody titer that Fig. 3 Listeria monocytogenes bacterium shadow immune mouse produces.
Detailed description of the invention
Below in conjunction with embodiment, the present invention will be further described, it should be understood that these embodiments are only used In the purpose of illustration, it is in no way intended to limit protection scope of the present invention.Food-borne Gram-positive is chosen in this part Bacterium Listeria monocytogenes (Listeria monocytogenes) is specifically described as embodiment.
Embodiment 1 prepares Listeria monocytogenes bacterium shadow
1. thalline is cultivated and is collected
With 1% (v/v) inoculum concentration, Listeria monocytogenes (Listeria monocytogenes) CMCC54004 (is purchased From Chinese industrial Culture Collection) seed liquor access through 121 DEG C of high pressure steam sterilizations BHI train Support base (purchased from Beijing overpass technical concern Co., Ltd), in 37 DEG C, incubated overnight under the conditions of 150r/min. Bacterium solution after cultivating is centrifugal 20min, supernatant discarded under the conditions of 1500r/min, uses SPSS (0.9%NaCl, analyzes pure, and reagent is purchased from Nanjing Chemistry Reagent Co., Ltd.) washing bacterial sediment.Washing Concrete grammar: with the resuspended thalline of SPSS, under the conditions of 1500r/min, centrifugal 20min, abandons Remove supernatant.Washing step is repeated 3 times.
2. prepared by bacterium shadow cell
(1) take the thalline after washing in step 1, be prepared as OD with sterilized water600Be 0.3~0.4 initial Bacteria suspension, adds the NaOH of final concentration of 27.500mmol/L, the SDS of final concentration of 0.234mmol/L (lauryl sodium sulfate), adds powder CaCO of 10.490mmol in every liter of initial bacteria suspension3, at 37 DEG C In shaking table, hatching 50min under shaking state, shaking speed is 100r/min.Then in 1500r/min bar Under part, centrifugal 20min, supernatant discarded, collect precipitation, uses SPSS according to method in step 1 Wash 3 times, resuspended with sterilized water, obtain re-suspension liquid A.Re-suspension liquid A volume and initial bacteria suspension volume phase Deng.NaOH, SDS add the most in form of an aqueous solutions to initial bacteria suspension.The NaOH aqueous solution dense Degree is 1.170mmol/L for the concentration of 137.5mmol/L, the SDS aqueous solution.OD600Refer to that solution is at 600nm Light absorption value at wavelength.
(2) in re-suspension liquid A, add the H of final concentration of 0.035% (mass percentage concentration)2O2, in 37 In DEG C shaking table, hatching 30min under shaking state, shaking speed is 100r/min.Then in 1500r/min Under the conditions of centrifugal 20min, supernatant discarded, collect precipitation, use sterile physiological salt according to method in step 1 Water washing precipitation 3 times, is then resuspended in 60% (v/v) (concentration expressed in percentage by volume) ethanol water, obtains Re-suspension liquid B.H2O2Add in form of an aqueous solutions to re-suspension liquid A, H2O2The percent mass of the aqueous solution is dense Degree is 0.175%.
(3) by re-suspension liquid B in 25 DEG C of shaking tables, hatch 30min under shaking state, shaking speed is 40 r/min.Then centrifugal 20min, supernatant discarded under the conditions of 1500r/min, collect precipitation, according to step In 1, method uses SPSS to wash 3 times, i.e. obtains Listeria monocytogenes bacterium shadow.
The preparation effect of embodiment 2 Listeria monocytogenes bacterium shadow
In detection embodiment 1, the survival rate of Listeria monocytogenes bacterium shadow prepared by method, is scanned electricity simultaneously Mirror (Scanning Electron Microscope, SEM), transmission electron microscope (Transmission Electron Microscope, TEM) observe.
(1) survival rate
At MSC-AdvantageTMIn two stage biological safety cabinet (purchased from Thermo Scientific), by above-mentioned Preparation bacterium shadow cell sterilized water carry out gradient dilution, extension rate be respectively 100 times, 101Again with 102 Times, then coat (purchased from Beijing overpass technical concern Co., Ltd) on BHI solid medium, in 37 DEG C EYELA constant incubator (purchased from Shanghai lover Instrument Ltd.) in cultivate 5d, every day observes flat Colony growth situation on plate.In triplicate.The results are shown in Table 1, as can be seen from Table 1 prepared by embodiment 1 Listeria monocytogenes bacterium shadow survival rate after cultivating 5d is 0%, i.e. exists without viable bacteria.
Table 1 Listeria monocytogenes bacterium shadow survival rate
Note: in table, data are means standard deviation.
(2) SEM observes
Listeria monocytogenes bacterium shadow SEM (SEM) observed result such as Fig. 1.Can be seen that because of thin Endochylema and nucleic acid are discharged, and the most morphologically Listeria monocytogenes bacterium shadow cell with Listeria monocytogenes living cells is Different.
(3) tem observation
TEM (transmission electron microscope) observed result of Listeria monocytogenes bacterium shadow is as in figure 2 it is shown, can see Cytoplasm has not been had in Listeria monocytogenes bacterium shadow.
The above results illustrates, embodiment 1 is successfully prepared Listeria monocytogenes bacterium shadow.
The immune effect inspection of embodiment 3 Listeria monocytogenes bacterium shadow
Use the Listeria monocytogenes bacterium shadow immune mouse that prepared by method in embodiment 1, exempt from investigating this bacterium shadow Whether Listeria monocytogenes antibody can be produced after epidemic disease.
Physiological saline is used to adjust the cell concentration of Listeria monocytogenes bacterium shadow to 1 × 108Cfu/mL, obtains immunity Reagent.Take 10 6 week old Balb/c mouse, every subcutaneous multi-point injection 2mL immunoreagent.Immunity knot Shu Hou, measures immune mouse antibodies in blood titer.Testing result is fig. 3, it is shown that implement Antibody can be produced after the Listeria monocytogenes bacterium shadow immune mouse that in example 1 prepared by method, after immune 4 weeks, In mouse blood, antibody titer is up to 10000, respond well.

Claims (10)

1. the preparation method of gram-positive bacteria bacterium shadow, it is characterised in that comprise the steps:
(1) in Gram-positive bacteria suspension, NaOH, SDS and CaCO are added3, hatch under the conditions of 30-40 DEG C, centrifugal, collect precipitation, after washing, make re-suspension liquid A;
(2) in described re-suspension liquid A, H is added2O2, hatch under the conditions of 30-40 DEG C, centrifugal, collect precipitation, after washing, make re-suspension liquid B;
(3) re-suspension liquid B is hatched under the conditions of 20-30 DEG C, centrifugal, collect precipitation, washing, obtain gram-positive bacteria bacterium shadow.
The preparation method of gram-positive bacteria bacterium shadow the most according to claim 1, it is characterized in that the final concentration of 20-40mmol/L of the NaOH added in Gram-positive bacteria suspension in step (1), the final concentration of 0.1-0.3mmol/L, CaCO in every liter of Gram-positive bacteria suspension of the SDS added3Addition be 9-11mmol;In step (1), shaking when suspension is hatched under the conditions of 80-120r/min, incubation time is 40-60min.
The preparation method of gram-positive bacteria bacterium shadow the most according to claim 2, it is characterised in that the H added in re-suspension liquid A described in step (2)2O2Final mass percentage concentration be 0.02-0.05%;In step (2), shaking when suspension is hatched under the conditions of 80-120r/min, incubation time is 20-40min.
The most according to claim 3, the preparation method of gram-positive bacteria bacterium shadow, it is characterised in that in step (3), shake when described re-suspension liquid B is hatched under the conditions of 30-50r/min, and incubation time is 20-40min.
5. according to the preparation method of one of claim 1-4 described gram-positive bacteria bacterium shadow, it is characterised in that use physiological saline to wash.
The preparation method of gram-positive bacteria bacterium shadow the most according to claim 5, it is characterised in that in step (2), re-suspension liquid B is that described precipitation is resuspended in gained in the ethanol water of 55-65%.
The preparation method of gram-positive bacteria bacterium shadow the most according to claim 6, it is characterised in that described re-suspension liquid A is that described precipitation is resuspended in gained after water.
8. according to the preparation method of one of claim 1-7 described gram-positive bacteria bacterium shadow, it is characterised in that described gram-positive bacteria is Listeria monocytogenes.
The preparation method of gram-positive bacteria bacterium shadow the most according to claim 8, it is characterised in that the OD of described Gram-positive bacteria suspension600It is 0.3~0.5.
10. the bacterium shadow that prepared by one of claim 1-9 method is in preparation application in terms of detecting the antibody of described gram-positive bacteria.
CN201610116309.6A 2016-03-01 2016-03-01 The preparation method and applications of gram-positive bacteria bacterium shadow Active CN105784984B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610116309.6A CN105784984B (en) 2016-03-01 2016-03-01 The preparation method and applications of gram-positive bacteria bacterium shadow

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610116309.6A CN105784984B (en) 2016-03-01 2016-03-01 The preparation method and applications of gram-positive bacteria bacterium shadow

Publications (2)

Publication Number Publication Date
CN105784984A true CN105784984A (en) 2016-07-20
CN105784984B CN105784984B (en) 2017-12-19

Family

ID=56387534

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610116309.6A Active CN105784984B (en) 2016-03-01 2016-03-01 The preparation method and applications of gram-positive bacteria bacterium shadow

Country Status (1)

Country Link
CN (1) CN105784984B (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109852563A (en) * 2019-03-06 2019-06-07 杨凌职业技术学院 A kind of chemical preparation process of eggs crack detection bacterium shadow
CN114921373A (en) * 2022-05-25 2022-08-19 中国农业科学院农业资源与农业区划研究所 Application of rhizobium oryzae in improving salt tolerance of plants

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101182538A (en) * 2007-11-26 2008-05-21 中国农业科学院哈尔滨兽医研究所 Bacillus coli perforating plasmid vector, construction method thereof and applications in bacterial ghost preparation
US20090239264A1 (en) * 2001-06-11 2009-09-24 Applied Nanosystems B.V. Immunization with bacterial ghost-based vaccines
CN101979504A (en) * 2010-10-26 2011-02-23 中国人民解放军军事医学科学院微生物流行病研究所 Yersinia pestis ghost, preparation method thereof, and application of Yersinia pestis ghost serving as vaccine
CN104232502A (en) * 2014-03-06 2014-12-24 北京市水产科学研究所 Bacterial ghost of aeromonas veronii as well as preparation method and application thereof
CN104805046A (en) * 2015-05-08 2015-07-29 齐鲁工业大学 Bacterial ghost preparation method of independent lysis gene E

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090239264A1 (en) * 2001-06-11 2009-09-24 Applied Nanosystems B.V. Immunization with bacterial ghost-based vaccines
CN101182538A (en) * 2007-11-26 2008-05-21 中国农业科学院哈尔滨兽医研究所 Bacillus coli perforating plasmid vector, construction method thereof and applications in bacterial ghost preparation
CN101979504A (en) * 2010-10-26 2011-02-23 中国人民解放军军事医学科学院微生物流行病研究所 Yersinia pestis ghost, preparation method thereof, and application of Yersinia pestis ghost serving as vaccine
CN104232502A (en) * 2014-03-06 2014-12-24 北京市水产科学研究所 Bacterial ghost of aeromonas veronii as well as preparation method and application thereof
CN104805046A (en) * 2015-05-08 2015-07-29 齐鲁工业大学 Bacterial ghost preparation method of independent lysis gene E

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
AMRO A. AMARA,ET AL: "Sponge-Like: A New Protocol for Preparing Bacterial Ghosts", 《THE SCIENTIFIC WORLD JOURNAL》 *
NAGARAJAN VINOD,ET AL: "Chemically induced Salmonella enteritidis ghosts as a novel vaccine candidate against virulent challenge in a rat model", 《VACCINE》 *
NAGARAJAN VINOD,ET AL: "Generation of a Novel Staphylococcus aureus Ghost Vaccine and Examination of Its Immunogenicity against Virulent Challenge in Rats", 《INFECTION AND IMMUNITY》 *
张玉敏 等: "菌影的制备方法及应用", 《动物医学进展》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109852563A (en) * 2019-03-06 2019-06-07 杨凌职业技术学院 A kind of chemical preparation process of eggs crack detection bacterium shadow
CN114921373A (en) * 2022-05-25 2022-08-19 中国农业科学院农业资源与农业区划研究所 Application of rhizobium oryzae in improving salt tolerance of plants

Also Published As

Publication number Publication date
CN105784984B (en) 2017-12-19

Similar Documents

Publication Publication Date Title
JP6095715B2 (en) Method for controlling the molecular weight of Streptococcus pneumoniae polysaccharide using carbon dioxide
JP5843612B2 (en) Method for controlling the molecular weight of Streptococcus pneumoniae serotype 19A polysaccharide
Wu et al. Production of bacterial ghosts from Gram-positive pathogen Listeria monocytogenes
CN104487086B (en) Non-animal derived nonalcoholic vaccine composition and preparation method thereof
CN103695508A (en) Fermentation and purification technologies of staphylococcus aureus HI recombinant protein
Prior et al. Bacterial-derived outer membrane vesicles are potent adjuvants that drive humoral and cellular immune responses
Tu et al. The role of macrophages in the protection of mice against leptospirosis: in vitro and in vivo studies
Corbel The serological relationship between Brucella spp., Yersinia enterocolitica serotype IX and Salmonella serotypes of Kauffmann-White group N
CN105784984A (en) Preparation method and application of gram-positive bacterium ghost
Sikarwar et al. Identification of Klebsiella pneumoniae by capsular polysaccharide polyclonal antibodies
CN105949300A (en) Method for extracting and separating and purifying protein
CN109847060A (en) Ox pasteurella multocida disease, Mannheimia haemolytica disease bivalent inactivated vaccine and preparation method thereof
Hatano et al. Immunogenic and antigenic properties of a heptavalent high-molecular-weight O-polysaccharide vaccine derived from Pseudomonas aeruginosa
Palmieri et al. GMMA as an alternative carrier for a glycoconjugate vaccine against group A Streptococcus
CN105682680A (en) Enhancing immunity to tuberculosis
Guo et al. Four simple biomimetic mineralization methods to improve the thermostability and immunogenicity of virus-like particles as a vaccine against foot-and-Mouth disease
CN101461940A (en) Photobacterium damsela vaccine as well as preparation method and use thereof
CN101014698A (en) Conserved inner core lipopolysaccharide epitopes as multi-species vaccine candidates
Li et al. Study on preparation of a Streptococcus suis ghost vaccine
CN107823638A (en) A kind of B group meningitis coccis restructuring chimeric protein vaccine and preparation method thereof
CN103933559B (en) Shigella multivalence combined vaccine
CN104448005B (en) The fusion rotein antigen of duck hepatitis A virus (HAV) 3 type VP1 albumen and LTB and application thereof
Shaw et al. Multicomponent vaccines against group A Streptococcus can effectively target broad disease presentations
CN103122336B (en) Goose parvovirus H-strain and application thereof in preventing and treating gosling plague
Dyatlov et al. Molecular Lipopolysaccharide Di-Vaccine Protects from Shiga-Toxin Producing Epidemic Strains of Escherichia coli O157: H7 and O104: H4

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant