CN109187964A - A kind of method and its application purifying detection simultaneously to avian leukosis and salmonellosis of chicken using one piece of hatching egg - Google Patents
A kind of method and its application purifying detection simultaneously to avian leukosis and salmonellosis of chicken using one piece of hatching egg Download PDFInfo
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- CN109187964A CN109187964A CN201811034968.0A CN201811034968A CN109187964A CN 109187964 A CN109187964 A CN 109187964A CN 201811034968 A CN201811034968 A CN 201811034968A CN 109187964 A CN109187964 A CN 109187964A
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- hatching egg
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- freeze thawing
- avian leukosis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
- G01N33/56916—Enterobacteria, e.g. shigella, salmonella, klebsiella, serratia
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
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Abstract
The invention belongs to animal and veterinary fields, and in particular to a kind of method and its application for purifying detection simultaneously to avian leukosis and salmonellosis of chicken using one piece of hatching egg.The method are as follows: breeder flock single cage is raised and numbered, and collects breeder flock primiparity hatching egg, and correspond with the number of Brood Laying hens；Hatching egg surface is carried out disinfection, and opens eggshell from hatching egg gas chamber end；It draws egg white and yolk is respectively placed in PE pipe and marks；Egg white and yolk sample are placed in refrigerator-freezer freeze thawing several times again；Egg white after taking freeze thawing detects ALV viral antigen by avian leukosis ELISA detection method；Physiological saline, chloroform is added in yolk after taking freeze thawing, after mixing on the oscillator, solution is centrifuged and is layered, lower layer is organic solvent, and middle layer is denatured lipoprotein, upper layer is the antibody extracted, draws the antibody on upper layer, detects antibodies toward salmonella by plate agglutination；Eliminate chicken group's individual of ALV viral antigen and the antibodies toward salmonella testing result positive.
Description
Technical field
The invention belongs to animal and veterinary fields, and in particular to a kind of to utilize one piece of hatching egg to avian leukosis and Salmonella gallinarum
Disease purifies the method and its application of detection simultaneously.
Background technique
Avian leukosis is that a kind of infectiousness caused by avian leukosis virus (Avian leukosis virus, ALV) is swollen
Tumor disease.It reports for the first time within 1908, the disease is in many country's generations and prevalence so far.China is to find this in broiler chicken group earliest
Case, in recent years, laying hen and indigenous chicken group case also persistently have been reported that, fall ill in rising trend.It is anti-that ALV belongs to reverse transcription section α
Retroviral category belongs to retrovirus with human immunodeficiency virus (HIV), and has similar viral genome knot
Structure.The virus can be divided into A~K totally 11 subgroups, wherein only A, B, C, D, J and K subgroup energy infected chicken.Pathogenic ALV with A,
B, based on this 4 kinds of subgroups of J and K, and popular most commonly used 4 exogenous virus subgroups in Chinese chicken group at present.The disease can
The different types of tumours such as lymphocytoma, fibrosarcoma, myelocytome, erythroblastoma, osteosclerosis, hemangioma are induced, are given
China's aviculture causes huge loss.
Avian leukosis is amplified step by step based on vertical transmission.Due to there is no effective vaccine to be prevented at present,
There is no suitable clinical treatment drug, on the prevention and control strategy of avian leukosis, the method for mainly taking purification detection superseded reaches
The purpose of purification.In conditional kind of chicken house, chicken group's purification can be carried out by detection meconium, blood plasma etc..Due to avian leukosis
Purification needs to acquire a large amount of sample and is detected, and most domestic kind chicken house there is no cell culture condition.
Chicken Sharpe door Salmonella disease always is (including white diarrhea, fowl typhoid and chicken paratyphoid) influences the bacillary of poultry husbandry
Infectious disease seriously affects one of the important hidden danger of survivability of chicks and the Adult Chicken underproduction, easily causes the pollution of poultry product,
To bring or cause food-safety problem.Salmonella is mostly important one of the borne Parasitic Encephalopathy cause of disease in the whole world, 30% food
Property germ contamination in source is by salmonellal, this is of great significance to public health.Salmonella gallinarum both can be passed vertically
It broadcasts, and can be with horizontal transmission.In actual production, usually quarantining by using blood plate agglutination test, it is next to eliminate positive chicken
Establish healthy group.Since blood plate agglutination test requires in advance that blood was collected, and a generation needs to carry out 2~3 times to adopt
Blood, it is larger to the stress reaction of chicken group, and operation of taking a blood sample is time-consuming and laborious, needs suitable skilled operator.Whole blood plate is solidifying
Collect the interference vulnerable to factors such as environment, light, operator's qualifications, and causes to judge by accident.Blood collection procedure is easy to generate chicken group
Stress, cause egg drop reduction or infectious disease.
The purification of current kind of chicken house avian leukosis and salmonellosis of chicken is all carried out separately, and the two does not carry out effectively
It is unified.
Summary of the invention
The present invention is in view of the deficiencies of the prior art, and it is an object of the present invention to provide a kind of husky to avian leukosis and chicken using one piece of hatching egg
The method and its application that door Salmonella disease is detected simultaneously.
For achieving the above object, the technical solution adopted by the present invention are as follows:
A method of detection being purified simultaneously to avian leukosis and salmonellosis of chicken using one piece of hatching egg, including is walked as follows
It is rapid:
(1) breeder flock single cage is raised and is numbered；Breeder flock primiparity hatching egg is collected, and a pair of with the number of Brood Laying hens one
It answers；
(2) hatching egg surface is carried out disinfection, and opens eggshell from hatching egg gas chamber end；It draws egg white and yolk is respectively placed in PE
In pipe and mark；Egg white and yolk sample are placed in refrigerator-freezer freeze thawing several times again；
(3) egg white after taking freeze thawing detects ALV viral antigen by avian leukosis ELISA detection method；
(4) physiological saline, chloroform is added in the yolk after taking freeze thawing, after mixing on the oscillator, solution is centrifuged and is layered,
Lower layer is organic solvent, and middle layer is denatured lipoprotein, and upper layer is the antibody extracted, draws the antibody on upper layer, solidifying by plate
Collection method detects antibodies toward salmonella；
(5) chicken group's individual of ALV viral antigen and the antibodies toward salmonella testing result positive is eliminated.
In above scheme, the temperature of the freeze thawing is -20 DEG C, and the number of freeze thawing is 1~3 time.
In above scheme, the yolk, physiological saline, chloroform volume ratio be 1:2:2.
In above scheme, the revolving speed of the centrifugation is 3500r/min, and the time of centrifugation is 15min.
Application of the above method in avian leukosis and salmonellosis of chicken purification context of detection.
Beneficial effects of the present invention are as follows: the present invention for the first time detects avian leukosis purification detection and salmonella purification very
Good combines, and effectively increases detection efficiency；The purification of avian leukosis and salmonellosis is carried out using same piece of hatching egg
Detection, compared with conventional purge detection method, reduces sampling number, is especially the reduction of blood sampling step, effectively prevents to chicken
Group stress, largely improve the utilization rate of hatching egg, and testing result determine quickly, it is accurate.
Specific embodiment
For a better understanding of the present invention, below with reference to the embodiment content that the present invention is furture elucidated, but it is of the invention
Content is not limited solely to the following examples.
Embodiment 1
A method of detection being purified simultaneously to avian leukosis and salmonellosis of chicken using one piece of hatching egg, including is walked as follows
It is rapid:
(1) acquisition and number of hatching egg: the breeder flock of quasi- purification is subjected to single cage raising, and to every breeder (or cage position)
It is numbered；The hatching egg of moment is collected as purification test sample, hatching egg is marked and is corresponded with chicken number；
(2) processing of sample: the hatching egg surface tincture of iodine and medicinal alcohol are sterilized in laboratory, with the micro shifting sterilized
Liquid device draws that 1ml egg white is placed in the PE pipe of 1.5ml, 1ml yolk is placed in the PE pipe of another 1.5ml, and makees respectively respectively
Good label, egg white and yolk are placed in -20 DEG C of refrigerator-freezer freeze thawing 1~3 time after the completion；Using preceding taking-up sample, restore to room temperature；
(3) egg white after taking freeze thawing is detected according to " avian leukosis antigen detection kit " method of IDEXX company
ALV viral antigen eliminates positive chicken according to positive threshold value standard；
(4) yolk after taking freeze thawing, by the ratio of yolk, physiological saline, chloroform 1:2:2 by volume, on the oscillator
After mixing, centrifuge is placed in 4 DEG C, 3500r/min centrifugation 15min, solution centrifugation layering, lower layer is organic solvent, middle layer
For denatured lipoprotein, upper layer is the antibody extracted, the antibody on upper layer is drawn, according to " the white diarrhea of China Veterinery Drug Inspection Office
Fowl typhoid multivalence Plate Antigen is referring to product operation instructions " to carry out " blood plate agglutination test " operation detection salmonella anti-
Body eliminates positive chicken according to kit Positive judgement standards；
(5) it eliminates ALV viral antigen and antibodies toward salmonella testing result is positive chicken group's individual.
Embodiment 2
Using 1 the method for embodiment, somewhere 30 pieces of hatching egg samples of kind chicken kind chicken house are detected, and scene is adopted
The blood for collecting corresponding chicken carries out antibodies toward salmonella detection using blood plate agglutination test, as a result as follows as control
Table:
1 ALV viral antigen of table and antibodies toward salmonella while testing result
Count avian leukosis and the positive number of salmonellosis of chicken detection: the positive number of avian leukosis antigen detection: 07,
12,14,23；Salmonella gallinarum antibody test positive number: 04,12,24, the positive number of contrast method detection: No. 12, eliminate institute
There is positive number chicken: as 04,07,12,14,23,24 all eliminate in this example.Illustrated by this test, Yolk antibody detection
Method is high compared with whole blood test method accuracy rate, false negative result caused by can be effectively reduced because of artificial judgement.
Obviously, above-described embodiment is only intended to clearly illustrate made example, and is not the limitation to embodiment.It is right
For those of ordinary skill in the art, can also make on the basis of the above description it is other it is various forms of variation or
It changes.There is no necessity and possibility to exhaust all the enbodiments.And the obvious variation or change therefore amplified
It moves within still in the protection scope of the invention.
Claims (5)
1. a kind of method for purifying detection simultaneously to avian leukosis and salmonellosis of chicken using one piece of hatching egg, which is characterized in that
Include the following steps:
(1) breeder flock single cage is raised and is numbered；Breeder flock primiparity hatching egg is collected, and is corresponded with the number of Brood Laying hens；
(2) hatching egg surface is carried out disinfection, and opens eggshell from hatching egg gas chamber end；It draws egg white and yolk is respectively placed in PE pipe
And it marks；Egg white and yolk sample are placed in refrigerator-freezer freeze thawing several times again；
(3) egg white after taking freeze thawing detects ALV viral antigen by avian leukosis ELISA detection method；
(4) physiological saline, chloroform is added in the yolk after taking freeze thawing, after mixing on the oscillator, solution is centrifuged and is layered, lower layer
For organic solvent, middle layer is denatured lipoprotein, and upper layer is the antibody extracted, draws the antibody on upper layer, passes through plate agglutination
Detect antibodies toward salmonella；
(5) chicken group's individual of ALV viral antigen and the antibodies toward salmonella testing result positive is eliminated.
2. the method according to claim 1, wherein the temperature of the freeze thawing is -20 DEG C, the number of freeze thawing is 1 ~
3 times.
3. the method according to claim 1, wherein the volume ratio of the yolk, physiological saline, chloroform is 1:2:
2。
4. the method according to claim 1, wherein the revolving speed of the centrifugation is 3500r/min, the time of centrifugation
For 15min.
5. one the method for claim 1 ~ 4 people is in the application of avian leukosis and salmonellosis of chicken purification context of detection.
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| CN201811034968.0A CN109187964A (en) | 2018-09-06 | 2018-09-06 | A kind of method and its application purifying detection simultaneously to avian leukosis and salmonellosis of chicken using one piece of hatching egg |
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| CN201811034968.0A CN109187964A (en) | 2018-09-06 | 2018-09-06 | A kind of method and its application purifying detection simultaneously to avian leukosis and salmonellosis of chicken using one piece of hatching egg |
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| CN109187964A true CN109187964A (en) | 2019-01-11 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112715478A (en) * | 2020-12-29 | 2021-04-30 | 北京市华都峪口禽业有限责任公司 | Method for purifying pullorum disease of egg-laying chicken and method for determining purification time of pullorum disease of egg-laying chicken |
| CN114532295A (en) * | 2022-03-19 | 2022-05-27 | 广西参皇养殖集团有限公司 | Method for jointly purifying leukemia and pullorum disease of high-quality chicken breeding core group |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112715478A (en) * | 2020-12-29 | 2021-04-30 | 北京市华都峪口禽业有限责任公司 | Method for purifying pullorum disease of egg-laying chicken and method for determining purification time of pullorum disease of egg-laying chicken |
| CN114532295A (en) * | 2022-03-19 | 2022-05-27 | 广西参皇养殖集团有限公司 | Method for jointly purifying leukemia and pullorum disease of high-quality chicken breeding core group |
| CN114532295B (en) * | 2022-03-19 | 2024-04-16 | 广西参皇养殖集团有限公司 | Combined purification method for leukemia and pullorum disease of high-quality chicken breeding core group |
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