CN106771274A - Brucella abortus reference serum storehouse - Google Patents
Brucella abortus reference serum storehouse Download PDFInfo
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- CN106771274A CN106771274A CN201610987222.6A CN201610987222A CN106771274A CN 106771274 A CN106771274 A CN 106771274A CN 201610987222 A CN201610987222 A CN 201610987222A CN 106771274 A CN106771274 A CN 106771274A
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Abstract
The present invention relates to a kind of Brucella abortus reference serum storehouse.Meaning of the present invention is：(1) take the lead in having built up Brucella abortus reference serum storehouse of certain scale in the world；(2) for the scientific evaluation of brucellosis diagnostic method provides cruel material；(3) all kinds of brucellosis diagnostic methods for exploitation for ox provide material base.
Description
Technical field
The present invention relates to a kind of Bovine brucellosis reference serum storehouse, belong to biological product technical field.
Background technology
Brucellosis (Brucellosis, abbreviation cloth disease) is drawn by brucella or brucella (Brucella)
The Amphixenosis being characterized with miscarrying and generating heat for rising, seriously threatens the life and health of people and many animals.This disease is not
But there is serious harm to the breeding and production performance of animal, it is often more important that, after people infection brucella, it tends to be difficult to control
More, so as to cause serious public health problem.Therefore in the country that brucella is popular, it is always public health to eliminate cloth disease
One of most important target in the works.
There is existing history remote in cloth disease, the mankind progressively deepen to the research understanding of the disease, cloth disease is examined in the world
Disconnected method is improved constantly in constantly improve, accuracy rate.Generally, brucella diagnostic method mainly includes diagnostic antigen method and resists
Body diagnosis, common laboratory mainly judges whether animal infects cloth Shandong by detecting the Brucella antibody in animal body
Salmonella.Brucella serological method not only includes traditional agglutination test (such as：The red antigen plate agglutination test of tiger, test tube resist
Former agglutination test, newborn ring agglutination test), also including sensitiveness is high, high specificity ELISA method, fluorescence polarization assay
(FPA), the new detection technique such as colloidal gold strip.
Agglutination test is a kind of conventional method of brucellosis diagnosis, including serum agglutination is tested, newborn ring precipitation is tried
Test and tested with human immunoglobulins, wherein classical standard tube agglutination test (STAT), plate agglutination test (PAT), with
Upper method is stopped using substantially in developed country, the substitute is buffering brucellergen agglutination test such as tiger red flat
Plate agglutination test (RBPT).The red Avian tubercula plain agglutination test antigen of tiger is through culture, inactivation, centrifugation with the good brucella bacterial strain of antigenicity
It is suspended in lactic acid buffer after being dyeed with brave red after collects thalline and is made, the method sensitivity is high, cheap, operation
Convenient, detection is quick, is suitable to the generaI investigation of animal population brucellosis.It is ox, sheep, traum's disease inspection in international trade
The specified experiment surveyed, the primary dcreening operation of people's brucellosis surveillance is also used in China.Newborn ring precipitation test is still cow cloth Lu Shi
The main method of bacterium monitoring, the method directly detects that screening can be carried out to milk cow colony, and the method is easy to be fast to cow's milk
Speed.
Unanimously think that complement fixation test (CFT) (CFT) is better than its other party in specificity in brucellosis patients inspection
Method, is often used to carry out qualitative inspection to tube agglutination test and rose bengal precipitation test test positive or suspicious case
Survey.Complement fixation test (CFT) is considered as always maximally effective cloth disease diagnostic method, can still be replaced without a kind of diagnostic method so far
Status of the complement fixation test (CFT) in serodiagnosis.But the preparation of the hemolysin required for complement fixation test (CFT) is difficult, experiment
It is cumbersome, it is difficult to clinically to commonly use, and the effect of GPC can be disturbed due to the complement in pig body, cause quick
Perception declines.
ELISA is a kind of diagnostic method suitable with CFT effects, and the method is easy to operate, and sensitivity is high, specificity compared with
It is good, the detection of a large amount of samples can be once completed, both animal population quarantine can be applied to as screening test, it is also used as
Confirmed diagnosis test.ELISA can be applied not only to the detection of serum antibody, apply also for the detection of antibody in milk sample.The cloth of ox
Shandong Salmonella disease ELISA detection method has been one of detection method for specifying in international trade, other kind of ELISA inspection of brucella
Survey method is also the focus of research.
At present, the serological method for being usually used in Bovine brucellosis detection mainly includes：Tube agglutination test (STAT), tiger
Red plate agglutination test ((RBPT), complement fixation test (CFT) (CFT), Brucella abortus indirect ELISA (iELISA) detection reagent
Box, brucella competitive ELISA (cELISA) antibody assay kit, fluorescence polarization (FPA) diagnostic method.Different serology are examined
Disconnected method respectively has its advantage and disadvantage, how the various serological diagnostic methods of objective evaluation specificity and sensitivity assessment, be unable to do without
Of certain scale, representative, background clearly serum sample storehouse.
Base has been established for Bovine brucellosis serological diagnostic method in Brucella abortus reference serum storehouse for evaluation is various
Plinth.
The content of the invention
The purpose of the present invention is by collecting or preparing of certain scale, certain representational, background clearly ox cloth
The serum of Shandong Salmonella difference Infection Status (natural infection, vaccine immunity, health), freezes and various Serologic detections through packing,
Brucella abortus reference serum storehouse is built up, for the scientific evaluation of brucellosis patients diagnostic method provides reference serum.
Technical scheme
1. Brucella abortus reference serum storehouse, it is characterised in that the Bovine brucellosis positive serum 30 containing natural infection
More than part；More than 30 parts of the Bovine brucellosis positive serum of vaccine immunity；More than 30 parts of brucellosis negative serum；Ox resists
The anti-yersinia enterocolitica O9 serum of Escherichia coli O 157 serum, ox, the anti-salmonella dublin serum of ox, ox are anti-thick
Each more than 2 parts of rough type brucella serum；This Serum Bank can be that the evaluation of all kinds of Bovine brucellosis serological diagnostic methods is carried
Material for reference, wherein：
(1) each part serum is accurately dispensed in Serum Bank, and lyophilized preservation, and every part of lyophilized sample number of serum is more than 10,
Meet the repeatability that different time is used；
(2) brucella positive serum in Serum Bank, brucella negative serum refer to rose bengal precipitation test,
The serum that the method such as tube agglutination test and complement fixation test (CFT) confirms jointly；
(3) Escherichia coli O 157, yersinia enterocolitica O9, salmonella berlin, Rough Anti-Brucella are easy
Cross reaction, the anti-Escherichia coli O 157 serum of ox, the anti-enterocolitis Yale of ox in Serum Bank are produced in Brucella antibody
Gloomy Salmonella O9 serum, the anti-salmonella berlin serum of ox, the anti-Rough Anti-Brucella blood of ox please be with corresponding bacterial cultures people
Work immune sheep is prepared from, and is used to evaluate the specificity of various serological diagnostic methods.
2. the application in the Brucella abortus reference serum storehouse described in claim 1, it is characterised in that use the Serum Bank
Reference serum can carry out scientific evaluation to the specificity of various ox cloth disease serological diagnostic method and sensitiveness.
The specific embodiment of the invention
The present invention uses the cow's serum for collecting different background source (natural infection, vaccine immunity, health population), by tiger
Red plate agglutination test (RBPT) Preliminary detection, then titration is carried out through tube agglutination test (SAT), and complement combines examination
Test (CFT) to make a definite diagnosis, dispense in a small amount after freezing, respectively as Brucella abortus positive reference serum, Brucella abortus vaccine immunity
Positive reference serum, Brucella abortus negative reference serum.
The hyper-immune serum of easily other Gram-negative bacterias of interference cloth disease serodiagnosis is prepared, mainly includes the anti-small intestine of ox
Colitis Yersinia ruckeri O9 positive serums, the anti-Escherichia coli O 157 positive serum of ox, the anti-salmonella berlin positive serum of ox,
The anti-Rough Anti-Brucella positive serum of ox.Above-mentioned serum is dispensed after freezing in a small amount, is referred to as brucella specific negative
Serum (Quality Control negative serum).
Above-mentioned background is understood, lyophilized serum as Brucella abortus reference serum storehouse is accurately dispensed, for ox cloth
Shandong Salmonella disease serological diagnostic method carries out characteristic and sensitivity assessment.This Serum Bank is various ox cloth disease serological diagnostic method
Evaluate and provide necessary material base.
1. the sick positive ox of clinically cloth is screened as positive serum samples sources.It is right respectively using rose bengal precipitation test
The cattle farm that there is abortion history Shandong District carries out cloth disease antibody test, to the positive serum for detecting, is tried with TA respectively
Testing carries out titration, while being made a definite diagnosis with complement fixation test (CFT).The ox positive to being diagnosed as cloth disease, gathers more serum,
And tube agglutination test redeterminates potency.The packing of each serum 0.5ml/ branch is lyophilized.
2. the immune cows antibody level of tracking and monitoring, gathers serum and comes as the positive serum sample that cloth disease vaccine is immunized
Source.Oral immunity, 50,000,000,000 CFU/ part are carried out to experiment milk cow using brucella disease vaccine (S2 plants).Adopted after the completion of immune
Antibody level is monitored weekly with rose bengal precipitation test, the serum of part sheep is collected in antibody level valency high-stage, according to facing
Bed positive serum sample preparation methods carry out titration and complement fixation test (CFT) is made a definite diagnosis.Each serum 0.5ml/ branch is dispensed and is frozen
It is dry.
3. monitoring healthy cow group cloth disease antibody situation, gathers the serum of negative antibody sheep as negative reference serum sample
Source.Antibody detection is carried out to miscarriage not occurring and the cattle farm of cloth disease vaccine not being immunized, selection rose bengal precipitation test,
Tube agglutination test and complement fixation test (CFT) are detected as the milk cow of feminine gender, prepare negative reference serum.By each serum 0.5ml/
Branch packing is lyophilized.
4. yersinia enterocolitica O9, Escherichia coli O 157, salmonella dublin, rough type cloth Lu Shi are prepared
Bacterium etc. easily produces the Gram-negative bacteria inactivation antigen of interference with smooth type brucella antiserum, and immunity test ox prepares phase
The antiserum answered, as Brucella abortus specific negative reference serum samples sources.The packing of each serum 0.5ml/ branch is lyophilized.
5. the background archives of above-mentioned various serum are set up.
The present invention relates to biomaterial resource information
(such as Xu Yunming is distinguished O9W22703 plants of yersinia enterocolitica (Yersinia.enterocolitica)
The design and analysis of detection yersinia enterocolitica and brucella LAMP primer, Chinese experimental diagnostics, 2013 1
The moon volume 17 the 1st phase, p19-22) come from Amphixenosis research institute of Jilin University；ETEC (Escherichia
Coli) O157 (CVCC1489), salmonella Dublin serotype (Salmonella sp.serotype dublin) C79-86
Strain (CVCC79351) and coarse bion brucella (Brucella melitensis Biotype roughness) M111
Strain (CVCC19) is all from National Veterinary Microbiological Culture Collection administrative center of China Veterinery Drug Inspection Office (see Chinese animal doctor
Medicine supervision institute, Chinese veterinary microorganism culture presevation administrative center write, Chinese animal doctor's strain catalogue (second edition), Chinese agriculture
Industry science tech publishing house, p33, p75, p109 in 2002).
Positive effect of the invention
The present invention relates to a kind of Brucella abortus reference serum storehouse, meaning of the present invention is：(1) take the lead in building in the world
Into Brucella abortus reference serum storehouse of certain scale；(2) for the scientific evaluation of cloth disease diagnostic method provides cruelty
Material；(3) all kinds of brucellosis diagnostic methods for exploitation for ox provide material base.
Embodiment
Following examples are not limited the invention to further illustrate the present invention.
Embodiment 1
--- the preparation of positive clinical reference serum
1. gathering clinic has part of cow serum more than 300 of abortion history and non-immune vaccine, is entered with rose bengal precipitation test
Row Preliminary Determination.Take the brucella rose bengal precipitation test antigen (lot number of serum 0.03ml to be checked and equivalent：201501),
Make flat board aggegation, in observing response result in 4min.Totally 79 parts of the positive serum that the red testing inspection of result tiger is arrived.
2. pair 79 parts of samples of the red test positive of tiger further determine Brucella antibody potency with tube agglutination test,
Retain antibody titer in the serum sample of 1 ﹕ more than 100, further carry out complement fixation test (CFT).Concrete operations are as follows：By test tube
0.5% phenol physiological saline of aggegation antigen (201501 batches) is (referred to as：Halite water) make 1:20 dilutions；Serum to be checked is made respectively
1:25、1:50、1:100、1:200 dilutions.Specific method is：First pipe first adds 2.4mL halite waters, often manages later and all adds
0.5mL halite waters, add 0.1mL serum to take 0.5mL and be added in second test tube after mixing and make 1 to first test tube:1 dilution.
Later each tubing is pushed away.First test tube should discard 1.5ml dilute serums；The brucella tube agglutination test of 20 times of dilutions is resisted
Former 0.5mL is separately added into each serology tubes, is shaken up, and puts 37 DEG C of 24h observation results.The dilution factor of last serum is 1:50、
1:100、1:200、1:400, while setting up opacity tube by table 1.Experiment sets positive serum (201501 batches), negative serum
(201501 batches) and antigen control (being shown in Table 1).
The tube agglutination test opacity tube of table 1 is prepared and transparency is represented
Result judgement：The result of determination on the premise of establishment is compareed.Serum titer to be checked is 1:100 (++) above persons are sun
Property；1:50 (++) are suspicious, are below feminine gender.
In 79 parts of positive serums of result, 68 parts of tube agglutination test potency are detected altogether higher than 1:100 serum sample.
3. rose bengal precipitation test and tube agglutination test detection are 68 parts of serum of the positive, physiological saline is used
Make 1:10 dilutions, after going out energy 30min in 56 DEG C, complement fixation test (CFT) operation are carried out by the art formula of table 2.Last time water-bath is heated
After 20min, standard haemolytic pipe is prepared by table 3, and carry out result judgement.
In 68 parts of positive serums of result, complement fixation test (CFT) detects 64 parts of positive serums altogether.
Each reagent pipetting volume amount of the complement fixation test (CFT) of table 2 and operation art formula
The standard haemolytic pipe of table 3 is prepared and criterion
Embodiment 2
--- the result that 79 parts of serum are determined using three kinds of different detection methods.
It is positive cow's serum through brave red primary dcreening operation to 79 parts, further carries out tube agglutination test and complement fixation test (CFT) inspection
Survey, three kinds of serum of the equal test positive of method of screening are accurately dispensed as Brucella abortus positive reference serum sample
It is lyophilized.Three kinds of distinct methods are following (table 4) to 79 parts of testing results of serum：
The result that the Brucella abortus positive serum of the clinical acquisitions of table 4 is determined using three kinds of different detection methods
Note：1.RBT：Rose bengal precipitation test, takes serum 0.03ml to be checked and is coagulated with the brucella red flat board of tiger of equivalent
Collection experiment antigen (lot number：201501) flat board aggegation, is made, in observing response result in 4min；++++：Occur big aggegation piece or
Particle, liquid is fully transparent；+++：There is obvious agglutinating particle, liquid is almost fully transparent；++：There is obvious aggegation
Grain, liquid is slightly transparent；+：Aggegation, opaque can slightly be seen；
—：Without aggegation, liquid is uniformly muddy.
2.SAT：Tube agglutination test, takes serum to be checked and distinguishes through 0.5% phenol physiological saline:25、1:50、1:100、1:
200 dilutions, with brucella tube agglutination test antigen (lot number：201501), reacted, 37 DEG C act on 22~24 hours,
Result of determination.4+：The complete aggegation of thalline and precipitation, liquid 100% are limpid；3+：Thalline almost complete aggegation and precipitation, liquid
75% is limpid；2+：There are significant aggegation and precipitation, liquid 50% is limpid；1+：There are clearly visible aggegation and precipitation, liquid
25% is limpid.
—：Without aggegation and precipitation, liquid bacterium solution is muddy.
3.CFT：Complement fixation test (CFT)；P：It is positive；N：It is negative.
4. cloth disease positive serum controls：2015-1 batches, loading amount：1ml, valid until on March 26th, 2025, from Chinese beast
Pharmaceuticals supervision institute cloth disease laboratory.
5. cloth disease negative serum control：2015-1 batches, loading amount：1ml, valid until on May 9th, 2025, from Chinese beast
Pharmaceuticals supervision institute cloth disease laboratory.
6. " B " represents the serum sample from ox.
Embodiment 3
--- the preparation of vaccine immune sera
The immune flock of sheep antibody level of tracking and monitoring, collection serum is originated as immuno positive serum sample.Using cloth disease epidemic disease
Seedling (S2 plants) carries out oral immunity, 2,500,000,000 CFU/ part to experiment sheep and goat.Brave red flat board aggegation is used after the completion of immune
Antibody level is monitored weekly in experiment, and part cow's serum is collected in antibody level high-stage, is prepared according to positive clinical serum sample
Method carries out titration and complement fixation test (CFT) is made a definite diagnosis.The packing of each serum 0.5ml/ branch is lyophilized.
Embodiment 4
--- the brucella negative reference serum (Quality Control negative serum) for easily intersecting with the generation of brucella antiserum
Prepare and quality inspection
1. Escherichia coli O 157 inactivation antigen, salmonella dublin inactivation antigen, yersinia enterocolitica O9
With the preparation of Rough Anti-Brucella M111 inactivation antigens
(1) be inoculated into for the Escherichia coli O 157 single bacterium colony of recovery and be equipped with by the preparation of Escherichia coli O 157 inactivation antigen
In the conical flask of 100ml ordinary broths (culture medium), 37 DEG C of shake culture 12-18h.10000r/min centrifugations 10min is collected
Thalline.With the resuspended thalline of 10mlPBS, 50 μ l formalin solutions are added, make content of formaldehyde be 0.2%, put 37 DEG C of inactivation 24h,
Rock 3-4 times midway.
(2) salmonella dublin, yersinia enterocolitica O9 use culture medium, antigens inactive method with it is big
Enterobacteria O157 is consistent；The culture medium that Rough Anti-Brucella M111 is used is pancreas peptone soybean broth culture medium (TSB), is resisted
Former ablation method is consistent with Escherichia coli O 157.
(3) the inactivation inspection of antigen：100 μ l are coated with LB solid mediums, put 37 DEG C of culture 18h, and observation whether there is bacterium life
It is long, if still there is bacterial growth to represent inactivation not exclusively inactivated, it is necessary to continue shake.If such as bacterial growth, representing that inactivation is complete.
2. prepared by immunizing antigen
(1) a small amount of Escherichia coli CVCC1489 plants of O157 inactivation antigen SPSSs are taken and is diluted to Maxwell opacity tube
Identical turbidity, with reference to the explanation of Maxwell opacity tube, now bacterial concentration is about 8 × 108CFU/ml, according to dilution ratio, calculating is gone out
Living bacterial liquid original concentration.Accordingly, the original antigen of preparation is adjusted to 4 × 1010CFU/ml, as immune use antigen, 2~8 DEG C
Save backup.
(2) ibid method that a small amount of salmonella berlin CVCC79351 plants of inactivation antigen is diluted into Maxwell opacity tube is identical turbid
Degree, with reference to the explanation of Maxwell opacity tube, now bacterial concentration is about 1 × 109CFU/ml, according to dilution ratio, calculates inactivated bacterial liquid
Original concentration.Accordingly, the original antigen of preparation is adjusted to 4 × 1010CFU/ml, used as immune use antigen, 2~8 DEG C of preservations are standby
With.
(3) ibid a small amount of yersinia enterocolitica W22703 plants of O9 inactivation antigen is diluted to Maxwell opacity tube by method
Identical turbidity, with reference to the explanation of Maxwell opacity tube, now bacterial concentration is about 8 × 108CFU/ml, according to dilution ratio, calculating is gone out
Living bacterial liquid original concentration.Accordingly, the original antigen of preparation is adjusted to 4 × 1010CFU/ml, as immune use antigen, 2~8 DEG C
Save backup.
(4) ibid a small amount of Rough Anti-Brucella M111 plants of inactivation antigen is diluted to the identical turbidity of Maxwell opacity tube by method,
With reference to the explanation of Maxwell opacity tube, now bacterial concentration is about 3 × 109CFU/ml, according to dilution ratio, calculates inactivated bacterial liquid original
Concentration.Accordingly, the original antigen of preparation is adjusted to 4 × 1010CFU/ml, used as immune use antigen, 2~8 DEG C save backup.
2. it is immunized
Immune general need to be carried out twice.Just exempt from：With 2ml inactivation antigens (4 × 1010CFU/ml) neck subcutaneous inoculation cloth disease
Negative ox.Booster immunization：After just exempting from 3 months, the subcutaneous booster immunization of doubling dosage neck 1 time.Take a blood sample within 2 weeks after booster immunization, use
Tube agglutination test determines agglutination titer and should be not less than 1:200.If potency does not reach standard, continue booster immunization 1~2 time, 15
Blood is tried again in the future, until potency is qualified.
3. prepared by serum
Two exempt from 14 after take a blood sample determine TA potency, if potency meet require, jugular vein blood collection, separate serum,
With 0.45 μm of filter filtration sterilization, proclin 300 to final concentration of 0.05% is added, it is aseptic subpackaged lyophilized.
4. check
Tested by following project to preparing the brucella for completing specificity Quality Control negative serum：
(1) each freeze-dried products color and luster of character observation, adds 1ml distilled water to observe its dissolubility.
(2) steriling test is by existing《Chinese veterinary pharmacopoeia》Annex is tested.
(3) specific assay
1) to the specific using each Quality Control negative serum as serum to be checked of brucella, brucella cELISA is used
Antibody assay kit is detected that calculating PI all should be less than 30%.
2) the specific 4 parts of Quality Control negative serums (BN1~BN4) to specific pathogen respectively make 1:100 dilutions, it is corresponding respectively
(concentration is 2 × 10 to inactivation antigen9CFU/ml tube agglutination test) is carried out, the positive should be judged to.
Embodiment 5.
--- brucella specific negative quality controlled serum quality test results
Reference method embodiment 4 prepares each brucella specific negative Quality Control blood sample, wherein BN1, BN2, BN3 and BN4
Immune Escherichia coli O 157 inactivation antigen, salmonella dublin inactivation antigen, Yersinia enterocolitica are represented number respectively
Antiserum prepared by bacterium O9 and Rough Anti-Brucella inactivation antigen.Negative quality controlled serum, after packing is lyophilized.Extract serum sample
Test, it is as a result as follows：
1. proterties is faint yellow loose agglomerate, can be dissolved rapidly after adding 1ml distilled water.
2. steriling test respectively extracts 5 bottles of negative serums at random, after being redissolved with 1ml sterile distilled waters, is transferred completely into 50ml
THIOGLYCOLLIC ACID salt (T.G) bottle in, to after culture in 37 DEG C of incubators 3 days after mixing, 0.2ml trainings are drawn from T.G bottles
Foster thing is inoculated with T.G tubules and each 2 of G.A inclined-planes respectively, and 1 is put 25 DEG C of cultures, and another is put 37 DEG C of cultures；Separately 0.2ml is taken to connect
A G.P tubule is planted, 25 DEG C of cultures are put, above test tube is cultivated 5.Records tests result (is shown in Table 5).
Specific negative quality controlled serum steriling test result in the Brucella abortus reference serum storehouse of table 5
"-" represents asepsis growth；"+" indicates bacteria growing.
3. specific assay
(1) to brucella specificity
Carried out using competitive ELISA method.Each Quality Control negative reference serum is tried with brucella cELISA antibody tests
Agent box is detected, is brucella negative serum less than 30% according to OD450 values calculating PI.Testing result is shown in Table 6.
The Brucella abortus Quality Control negative serum specific assay result of table 6
(2) to the specificity of specific pathogen
3 bottles of serum are respectively randomly selected from Quality Control negative serum BN1, BN2, BN3, BN4, is answered with 1m sterile distilled waters respectively
It is molten, by each redissolution serum with corresponding tube agglutination test antigen, tube agglutination test and result judgement are carried out, assay is shown in Table
7。
The brucella specific negative quality controlled serum TA potency assay of table 7
Note：4+：The complete aggegation of thalline and precipitation, liquid 100% are limpid；3+：Thalline almost complete aggegation and precipitation, liquid
75% is limpid；2+：There are significant aggegation and precipitation, liquid 50% is limpid；1+：There are clearly visible aggegation and precipitation, liquid
25% is limpid；—：Without aggegation and precipitation, liquid bacterium solution is muddy.2++:The intensity of agglutination is between 3+ and 2+, but closer 2+；2
+-:The intensity of agglutination is between 2+ and 1+, but closer 2+.
Claims (2)
1. Brucella abortus reference serum storehouse, it is characterised in that 30 parts of the Bovine brucellosis positive serum containing natural infection with
On；More than 30 parts of the Bovine brucellosis positive serum of vaccine immunity；More than 30 parts of cloth brucella negative serum；The anti-large intestine of ox
The anti-yersinia enterocolitica O9 serum of bacillus O157 serum, ox, the anti-salmonella berlin serum of ox, the anti-rough type cloth of ox
Each more than 2 parts of Shandong Salmonella serum；This Serum Bank can provide reference for the evaluation of all kinds of Bovine brucellosis serological diagnostic methods
Material, wherein：
(1) each part serum is accurately dispensed in Serum Bank, and lyophilized preservation, and every part of lyophilized sample number of serum is more than 10, meets
The repeatability that different time is used；
(2) brucella positive serum in Serum Bank, brucella negative serum refer to rose bengal precipitation test, test tube
The serum that the method such as agglutination test and complement fixation test (CFT) confirms jointly；
(3) Escherichia coli O 157, yersinia enterocolitica O9, salmonella berlin, Rough Anti-Brucella are easy to cloth
Shandong Salmonella antibody produces cross reaction, the anti-Escherichia coli O 157 serum of ox in Serum Bank, the anti-Yersinia enterocolitica of ox
The anti-salmonella berlin serum of bacterium O9 serum, ox, the anti-Rough Anti-Brucella Serum of ox are manually exempted from corresponding bacterial cultures
Sheep is prepared from epidemic disease, is used to evaluate the specificity of various serological diagnostic methods.
2. the application in the Brucella abortus reference serum storehouse described in claim 1, it is characterised in that use the reference of the Serum Bank
Serum can carry out scientific evaluation to the specificity and sensitiveness of various Bovine brucellosis serological diagnostic methods.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610987222.6A CN106771274A (en) | 2016-11-10 | 2016-11-10 | Brucella abortus reference serum storehouse |
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| CN115508561A (en) * | 2022-08-02 | 2022-12-23 | 中国农业科学院北京畜牧兽医研究所 | Method and kit for evaluating whether cattle are immunized with brucellosis vaccine |
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| CN105067812A (en) * | 2015-08-06 | 2015-11-18 | 中国兽医药品监察所 | Bovine brucella indirect ELISA antibody detection kit |
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| CN105067812A (en) * | 2015-08-06 | 2015-11-18 | 中国兽医药品监察所 | Bovine brucella indirect ELISA antibody detection kit |
Non-Patent Citations (2)
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| 王芳等: "牛布鲁氏菌间接ELISA 抗体检测方法的建立", 《中国农业科学》 * |
| 赵仲堂: "《流行病学研究方法与应用》", 31 August 2000, 科学出版社 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115508561A (en) * | 2022-08-02 | 2022-12-23 | 中国农业科学院北京畜牧兽医研究所 | Method and kit for evaluating whether cattle are immunized with brucellosis vaccine |
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