CN109541239A - A method of based on parathormone blood biochemical markers breeding Wuding Chicken - Google Patents
A method of based on parathormone blood biochemical markers breeding Wuding Chicken Download PDFInfo
- Publication number
- CN109541239A CN109541239A CN201811572705.5A CN201811572705A CN109541239A CN 109541239 A CN109541239 A CN 109541239A CN 201811572705 A CN201811572705 A CN 201811572705A CN 109541239 A CN109541239 A CN 109541239A
- Authority
- CN
- China
- Prior art keywords
- chicken
- wuding
- wuding chicken
- skeleton development
- pth
- 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
Links
Classifications
-
- 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
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
- G01N33/78—Thyroid gland hormones, e.g. T3, T4, TBH, TBG or their receptors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Endocrinology (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention relates to a kind of methods based on parathormone blood biochemical markers breeding Wuding Chicken, belong to poultry blood biochemical markers technical field, the present invention specifically includes step: 1) acquiring the blood sample of Wuding Chicken to be determined, prepare serum, detect the concentration value of its parathyroid hormone (PTH)；2) concentration value that step 1) measures is compared with Wuding Chicken blood biochemical markers, Wuding Chicken to be measured is distinguished as skeleton development normal type Wuding Chicken and skeleton development ectype Wuding Chicken.Concentration variation of the present invention by measurement parathyroid hormone (PTH) in skeleton development normal type and skeleton development ectype Wuding Chicken, bone density, bone strength in conjunction with the Wuding Chicken of detection, give skeleton development normal type and skeleton development ectype Wuding Chicken blood biochemical markers, screening high-quality chicken, it provides fundamental basis for the early stage breeding of Wuding Chicken, also it is the selection for probing into Wuding Chicken, provides new approaches in breeding work for Yunnan local chicken breeds.
Description
Technical field
The invention belongs to poultry blood biochemical markers technical fields, specifically, are related to a kind of based on parathormone blood
The method of liquid biochemical marker breeding Wuding Chicken.
Background technique
For bone as organ important in poultry body, function is to play support, protection and movement to the body of poultry
Effect.And a nearly century comes, the breeding of poultry high intensity production performance has a negative impact to poultry skeleton development, causes to increase
The disease incidence for adding the osteopathy such as skeleton development defect, osteoporosis, osteomalacia affects the sound development of domestic fowl farming.Family chicken bone is close
Degree and intensity are the important economic characters of family chicken, every year because the osteopathy such as skeleton development defect, osteoporosis, osteomalacia give family chicken industry
Cause huge economic loss.American Studies show that the death rate caused by skeleton deformity accounts for about 1.5%-3.2%, show bone
Deformity brings hundred million dollars of about 15-20 of loss (Jendral et al., 2008) to aviculture every year.China's broiler chicken is averaged osteopathy
Disease incidence is 3%-16%, and direct economic loss is about 50-80 hundred million.Wherein, since heredity and trophic factor cause skeleton development
Defect and cause the osteoporosis, osteomalacia, thigh bone of poultry the generations of osteopathy such as to become fragile, cause the huge economic damage of domestic fowl farming
It loses.
Parathyroid hormone (parathyroid hormone, PTH) is synthesized and is secreted by parathyroid gland, by 84 amino
Sour polypeptide composition, molecular weight 9500u, main function are to maintain the balance of alcium and phosphor metabolization, and main target organ is bone, kidney
Dirty and small intestine.PTH has the function of the pros and cons in the homeostasis process of bone metabolism, and the use of discontinuity PTH promotes
Bon e formation；The use of continuity PTH will induce bone resorption.
Wuding Chicken mainly originates in the Wuding county of Yunnan Province Chuxiong autonomous prefecture, and figure is larger, and delicious meat is strong and brisk in taste, standby
Liked by people, becomes exclusive one " business card " in Wuding county or even Yunnan Province.Not yet in relation to parathyroid hormone to Wuding Chicken
The research that skeleton development influences, to meet the needs of people are to Local chicken breeds, it is necessary to propose that a kind of energy breeding skeleton development is good
The new method of good Wuding, Yunnan chicken establishes a blood biochemical markers by the research of parathyroid hormone and skeleton development,
It lays the groundwork for the early stage breeding of Wuding, Yunnan chicken.
Summary of the invention
In order to solve the problems, such as background technique, the present invention provides one kind to be based on parathormone blood biochemical mark
The method for remembering breeding Wuding Chicken, by measurement parathyroid hormone (PTH) in skeleton development normal type and skeleton development ectype
Concentration variation in Wuding Chicken, bone density, bone strength in conjunction with the Wuding Chicken of detection give skeleton development normal type and bone
Dysplastic type Wuding Chicken blood biochemical markers, screening high-quality chicken are provided fundamental basis for the early stage breeding of Wuding Chicken, also for
The selection of Wuding Chicken is probed into, provides new approaches in working in the rain for Yunnan local chicken breeds.
To achieve the above object, the present invention is achieved through the following technical solutions:
The method based on parathormone blood biochemical markers breeding Wuding Chicken, specifically includes the following steps:
1) blood sample for acquiring Wuding Chicken to be determined, prepares serum, detects the concentration value of its parathyroid hormone (PTH)；
2) concentration value that step 1) measures is compared with Wuding Chicken blood biochemical markers, Wuding Chicken to be measured is distinguished as
Skeleton development normal type Wuding Chicken and skeleton development ectype Wuding Chicken.
Further, Wuding Chicken blood biochemical markers establishment specifically includes the following steps:
A. skeleton development normal type Wuding Chicken family and skeleton development ectype are randomly choosed from Wuding Chicken sources group
Wuding Chicken family detects its thigh bone bone density, bone strength respectively, acquires blood sample, prepares serum；
B. serum parathyroid hormone (PTH) content assaying method: parathormone (PTH) ELISA reagent is selected
Box, chicken parathyroid hormone (PTH) content in competitive enzyme-linked immune method detection assay sample, first with anti-PTH antibody packet
By microwell plate, it is prepared into solid phase antibody, sample, biotin labeling PTH and horseradish peroxidase-labeled to be measured is then added
Avidin (HRP) is allowed to form coated antibody-biotinylation PTH- Avidin (HRP) immune complex, after thoroughly washing
Substrate TMB is added to develop the color.The PTH amount in the binding capacity and sample of PTH is marked to be inversely proportional.In microplate reader 450nm wavelength after developing the color
Lower measurement absorbance (OD value) is fitted concentration-absorbance standard curve by mapping, and inverse goes out chicken parathyroid gland in sample to be tested
Hormone (PTH) content；
C. according to step b the data obtained, skeleton development normal type Wuding Chicken and skeleton development ectype Wuding Chicken first are obtained
The concentration value of parathyrine by shape carries out parathormone and bone is close further according to the thigh bone bone density of selected Wuding Chicken, bone strength
The correlation analysis of degree, bone strength；
D. according to the obtained analysis of step c as a result, determining the blood biochemical markers of Wuding Chicken.
Further, the blood biochemical markers of the Wuding Chicken are the concentration value of parathormone, when a monthly age, when first shape
For the concentration value of other parathyrine in 167.28pg/ml hereinafter, being determined as skeleton development normal type Wuding Chicken, on the contrary is that skeleton development is abnormal
Type Wuding Chicken；When two monthly ages, when parathormone concentration value in 172.57pg/ml hereinafter, being determined as skeleton development normal type
Wuding Chicken, on the contrary it is skeleton development ectype Wuding Chicken.
Further, it cuts off the water supply before blood sampling, is 12 hours jejunitas, carrying out wing venous blood sampling the next morning；It is collected with centrifuge tube
Blood sample, standing 30min, 3000rpm is centrifuged 20min at room temperature, takes supernatant, obtains serum.
Beneficial effects of the present invention:
The present invention is by measurement parathyroid hormone (PTH) in skeleton development normal type and skeleton development ectype Wuding Chicken
In concentration variation, bone density, bone strength in conjunction with the Wuding Chicken of detection, give skeleton development normal type and skeleton development is different
Normal type Wuding Chicken blood biochemical markers, screening high-quality chicken are provided fundamental basis for the early stage breeding of Wuding Chicken, also to probe into force
Determine the selection of chicken, provides new approaches in working in the rain for Yunnan local chicken breeds.
Specific embodiment
It, below will be to preferred reality of the invention in order to keep the purpose of the present invention, technical scheme and beneficial effects clearer
It applies example to be described in detail, to facilitate the technical staff to understand.
Embodiment 1
1. the acquisition and preservation of laboratory sample
(1) preparation of serum
A monthly age is randomly selected in skeleton development normal type Wuding Chicken family and skeleton development ectype Wuding Chicken family
With two monthly age skeleton development normal types and skeleton development ectype Wuding Chicken each 20, its blood measuring bone metabolism phase is acquired respectively
Close biochemical indicator.It cuts off the water supply before blood sampling, is 12 hours jejunitas, carrying out wing venous blood sampling the next morning.Blood is collected with 10ml centrifuge tube
Sample 3-4ml, standing 30min, 3000rpm is centrifuged 20min at room temperature, takes supernatant (serum) in the centrifuge tube of 1.5ml, puts
Enter and is saved backup in -20 DEG C of refrigerators.
(2) acquisition of thigh bone sample
20 are respectively butchered respectively at 1,2 monthly age skeleton development normal types and skeleton development ectype Wuding Chicken, acquisition test
The femur and shin bone of the left and right leg of chicken.The soft tissue and connective tissue for removing bone surface, it is dry with 0.75% normal saline flushing
Only, and with the gauze for being moistened with physiological saline wrap up, be tamping with valve bag, -20 DEG C save backup in measurement bone density and bone strength
Index.
2. thigh bone Bone mineral density
After test chicken left femur and left tibia thaw, using QDR-4500 type Dual-energy X-rays absorptionmetry (Hologic company,
The U.S.) measurement bone density.
3. thigh bone bone strength detects
After test chicken left femur and left tibia thaw, by bone samples be placed in electronic universal tester (AG-IS, day the island proper
Saliva company) under carry out three point bending test, to detect bone strength index: elasticity modulus, yield stress, bursting stress, fracture are answered
Power.Span calculation formula used by this test are as follows: L=(a+3b) ± 0.5b, wherein a is the diameter of supporting rail, and b is bone
Diameter.Loading velocity is 2mm/min.Detection unit is Kunming University of Science and Technology's Materials Academy experimental center.
4. blood parameters detect
Serum parathyroid hormone (PTH) content assaying method: the measuring method of parathormone (PTH) is according to kit
The operating method that production firm (Nanjing Bo Pai Bioisystech Co., Ltd (U.S. R&D dispenses kit)) operation instructions provide
It carries out:
With chicken parathyroid hormone (PTH) content in competitive enzyme-linked immune method detection assay sample is used, first with anti-PTH
Antibody is coated with microwell plate, is prepared into solid phase antibody, sample, biotin labeling PTH and horseradish peroxidase to be measured is then added
Enzyme marks Avidin (HRP), is allowed to form coated antibody-biotinylation PTH- Avidin (HRP) immune complex, by thorough
After washing plus substrate TMB develops the color.The PTH amount in the binding capacity and sample of PTH is marked to be inversely proportional.In microplate reader after developing the color
Absorbance (OD value) is measured under 450nm wavelength, concentration-absorbance standard curve is fitted by mapping, inverse goes out in sample to be tested
Chicken parathyroid hormone (PTH) content.Concrete operations are carried out referring to kit specification.
5. data statistics
1 one monthly age of table Wuding Chicken bone density, bone strength data
Note: colleague's data, " * " indicate significant difference (P < 0.05).
2 two monthly age of table Wuding Chicken bone density, bone strength data
Note: colleague's data, " * " indicate significant difference (P < 0.05).
The content of 3 one monthly age of table Wuding Chicken serum parathyroid hormone
Note: colleague's data, " * " indicate significant difference (P < 0.05).
The content of 4 two monthly age of table Wuding Chicken serum parathyroid hormone
5 one monthly age of table skeleton development large fowl serum parathyroid hormone and bone density, bone strength correlation analysis
Note: " * * " indicates that, in 0.01 horizontal significant correlation, " * " is indicated in 0.05 horizontal significant correlation.
6 one monthly age of table skeleton development ectype chicken serum parathyroid hormone and bone density, bone strength correlation analysis
Note: " * * " indicates that, in 0.01 horizontal significant correlation, " * " is indicated in 0.05 horizontal significant correlation.
7 two monthly age of table skeleton development large fowl serum parathyroid hormone and bone density, bone strength correlation analysis
Note: " * * " indicates that, in 0.01 horizontal significant correlation, " * " is indicated in 0.05 horizontal significant correlation.
8 two monthly age of table skeleton development ectype chicken serum parathyroid hormone and bone density, bone strength correlation analysis
Note: " * * " indicates that, in 0.01 horizontal significant correlation, " * " is indicated in 0.05 horizontal significant correlation.
6. interpretation of result
By Tables 1 and 2 it is found that bone density data when one, two monthly age: the femur bone of skeleton development normal type Wuding Chicken is close
Degree content, shin bone bone density are above skeleton development ectype Wuding Chicken, wherein two time differences at monthly age are different significant (P < 0.05)；
One, bone strength data when two monthly ages: femur backbone elasticity modulus, the tibial bone dry elasticity mould of skeleton development normal type Wuding Chicken
Amount, femur backbone yield stress, shin bone backbone yield stress, femur backbone maximum stress, shin bone backbone maximum stress, femur bone
Dry fracture strength, shin bone backbone fracture strength are all remarkably higher than skeleton development ectype Wuding Chicken (P < 0.05).
By table 3 and table 4 it is found that PTH content data in Wuding County's chicken serum when one, two monthly age: skeleton development normal type is military
Determine PTH content in chicken serum and be lower than skeleton development ectype Wuding Chicken, a time difference at monthly age is different significant (P < 0.05).
By the correlation analysis of one, two monthly age Wuding Chicken bone densities and bone strength it is found that when one, two monthly age, bone hair
Educate the femoral bmd content of normal type Wuding Chicken and skeleton development ectype Wuding Chicken with shin bone bone density, femur bone
Dry elasticity modulus, tibial bone dry elasticity modulus, femur backbone yield stress, shin bone backbone yield stress, femur backbone maximum are answered
Power, shin bone backbone maximum stress, femur backbone fracture strength, shin bone backbone fracture strength are positively correlated and (are shown in Table 5, table 6, table
7, table 8).By the correlation analysis of PTH content in one, two monthly age Wuding Chicken serum and bone density and bone strength it is found that one, two
When the monthly age, PTH content is close with femur bone in the serum of skeleton development normal type Wuding Chicken and skeleton development ectype Wuding Chicken
Spend content, shin bone bone density, femur backbone elasticity modulus, tibial bone dry elasticity modulus, femur backbone yield stress, shin bone
Backbone yield stress, femur backbone maximum stress, shin bone backbone maximum stress, femur backbone fracture strength, the fracture of shin bone backbone
Stress is negatively correlated (being shown in Table 5, table 6, table 7, table 8), wherein in serum PTH content and femoral bmd related coefficient-
0.578~-0.830, with shin bone bone density related coefficient -0.397~-0.754；It is obtained by above-mentioned correlation data result,
PTH content has decisive role to early stage Wuding Chicken skeleton development in serum, can be used as the excellent skeleton development Wuding County of early stage breeding
The blood biochemical markers of chicken.
Embodiment 2
In the monthly age Wuding Chicken 100 that Wuding, Yunnan and farm select uniformity of type, are in a good state of health, examine respectively
PTH concentration value in its blood is surveyed, statistical packet is carried out according to gained concentration value in a certain range, obtains following statistical form.
The content of 9 one monthly age of table Wuding Chicken PTH counts
By this 5 group of monthly age Wuding Chicken in strict accordance with immune programme, raised under identical rearing conditions to two monthly ages, difference
Its shin bone bone density (being detected using portable borne densitometers) is measured, each group statistical result is following (being shown in Table 10).
10 1 monthly age of table Wuding Chicken mean tibial bone density statistics
As seen from Table 10, third group mean tibial bone density is compared with first group and second group of significant decrease, and fourth, fifth group
Successively reducing, the interval range for reasoning out third group is a significant break range, i.e. when a monthly age, PTH content in serum
Range (160.26-170.25pg/ml) is break range.
Above-mentioned steps are being repeated to the chicken within the scope of the break, finally obtaining 167.28pg/ml is that PTH contains in serum
The significant break of amount.
The PTH content in Wuding County's chicken serum at above-mentioned two monthly age is detected, as a result following (being shown in Table 11)
The content of 11 2 monthly age of table Wuding Chicken PTH counts
By this 5 group of two monthly age Wuding Chicken in strict accordance with immune programme, raised under identical rearing conditions to three monthly ages, difference
Its shin bone bone density (wherein there are 2 Wuding Chickens because of leg joint injury, to be not counted in statistical result) is measured, each group statistical result is as follows
(being shown in Table 12).
The Wuding Chicken mean tibial bone density accrual accounting of 12 3 monthly age of table
As seen from Table 12, third group mean tibial bone density increment two groups of significant decreases earlier above, and the 4th group and the 5th
Group Wuding Chicken mean tibial bone density increment reduces, and reasoning out 3rd interval range is significant a break range, i.e., two
When the monthly age, PTH content (170.01-174.00pg/ml) is break range in serum.
By within the scope of two breaks Wuding Chicken and adjacent interval in PTH concentration value in 170.01-174.00pg/ml
Chicken combination, then repeat above-mentioned steps, finally obtaining 172.57pg/ml is PTH content in serum
Significant break.
In Yunnan, experiment, conclusion and above-mentioned experiment knot is repeated several times by the above experimental procedure in other Wuding Chicken farms
By identical, conclusion are as follows: when a monthly age, PTH content range (160.26-170.25pg/ml) is significant break range in serum,
And 167.28pg/ml is the significant break of PTH content in serum；When two monthly ages, PTH content (170.01- in serum
It 174.00pg/ml) is break range, and 172.57pg/ml is the significant break of PTH content in serum.
Embodiment 3
1. the preparation of experimental material
In Wuding, Yunnan county, each Wuding Chicken farm sampling one monthly age Wuding Chicken totally 120 (includes skeleton development normal type
With skeleton development ectype), two monthly age Wuding Chickens totally 120 (include skeleton development normal type and skeleton development ectype), point
Its shin bone bone density is not measured；Wing venous blood sampling is carried out, blood sample is collected with 5ml centrifuge tube, stands 30min at room temperature
3000rpm is centrifuged 20min, takes supernatant, extremely serum.
2. detecting the concentration value of PTH in serum
It carries out strictly regulating operation according to PTH ELISA (ELISA) kit, be measured using double antibody sandwich method
The level of chicken osteocalcin (PTH) in sample is measured absorbance (OD value) under 450nm wavelength with microplate reader, passes through standard curve
Calculate osteocalcin (PTH) content in sample.
3. comparative analysis
When one monthly age, shin bone bone density is in 4.4176g/cm2Below is 31, in 0.4356g/cm2It is above shared
72, i.e. in 120 Wuding Chickens, skeleton development ectype Wuding Chicken 31, skeleton development normal type Wuding Chicken 72.At 31
In skeleton development ectype Wuding Chicken, there is PTH content in 27 Wuding County's chicken serums to account for skeleton development in 167.28pg/ml or more
The 87.10% of ectype Wuding Chicken.In 72 skeleton development normal type Wuding Chickens, there is PTH content in 65 Wuding County's chicken serums
In 167.28pg/ml hereinafter, accounting for the 90.28% of skeleton development normal type Wuding Chicken.
When two monthly ages, shin bone bone density is in 0.4066g/cm2Below is 33, in 0.5124g/cm2It is above shared
76, i.e. in 120 Wuding Chickens, skeleton development ectype Wuding Chicken 33, skeleton development normal type Wuding Chicken 76.At 33
In skeleton development ectype Wuding Chicken, there is PTH content in 26 Wuding County's chicken serums to account for skeleton development in 172.57pg/ml or more
The 78.79% of ectype Wuding Chicken.In 76 skeleton development normal type Wuding Chickens, there is PTH content in 70 Wuding County's chicken serums
In 172.57pg/ml hereinafter, accounting for the 92.11% of skeleton development normal type Wuding Chicken.
In conclusion the present invention is by PTH content in measurement blood in skeleton development normal type and skeleton development ectype
Concentration variation in Wuding Chicken, Wuding Chicken bone density, bone strength index in conjunction with measurement give skeleton development normal type and bone
Blood biochemical markers in bone dysplastic type Wuding Chicken, screening high-quality chicken are provided fundamental basis, together for the early stage breeding of Wuding Chicken
When be also to explore the high-quality indigenous a breed of chicken new method in plateau, new approaches are provided in breeding work for Yunnan local chicken breeds.
Finally, it is stated that preferred embodiment above is only used to illustrate the technical scheme of the present invention rather than limits, although logical
It crosses above preferred embodiment the present invention is described in detail, however, those skilled in the art should understand that, can be
Various changes are made to it in form and in details, without departing from claims of the present invention limited range.
Claims (4)
1. a kind of method based on parathormone blood biochemical markers breeding Wuding Chicken, it is characterised in that: specifically include following
Step:
1) blood sample for acquiring Wuding Chicken to be determined, prepares serum, detects the concentration value of its parathyroid hormone (PTH)；
2) concentration value that step 1) measures is compared with Wuding Chicken blood biochemical markers, Wuding Chicken to be measured is distinguished as bone
Develop normal type Wuding Chicken and skeleton development ectype Wuding Chicken.
2. a kind of method based on parathormone blood biochemical markers breeding Wuding Chicken according to claim 1, special
Sign is: the establishments of Wuding Chicken blood biochemical markers specifically includes the following steps:
A. skeleton development normal type Wuding Chicken family and skeleton development ectype Wuding County are randomly choosed from Wuding Chicken sources group
Chicken family detects its thigh bone bone density, bone strength respectively, acquires blood sample, prepares serum；
B. serum parathyroid hormone (PTH) content assaying method: parathormone (PTH) enzyme linked immunological kit is selected, is adopted
With chicken parathyroid hormone cellulose content in competitive enzyme-linked immune method detection assay sample, microwell plate, system are coated with anti-PTH antibody first
It is standby that sample, biotin labeling PTH and Horseradish peroxidase-conjugated avidin (HRP) to be measured is then added at solid phase antibody,
It is allowed to form coated antibody-biotinylation PTH- Avidin (HRP) immune complex, after thoroughly washing plus substrate TMB is aobvious
Color；The PTH amount in the binding capacity and sample of PTH is marked to be inversely proportional；Absorbance is measured under microplate reader 450nm wavelength after developing the color
(OD value) is fitted concentration-absorbance standard curve by mapping, and inverse goes out chicken parathyroid hormone cellulose content in sample to be tested；
C. it according to step b the data obtained, obtains by skeleton development normal type Wuding Chicken and skeleton development ectype Wuding Chicken first shape
The concentration value of parathyrine, further according to the thigh bone bone density of selected Wuding Chicken, bone strength, carry out parathormone and bone density,
The correlation analysis of bone strength；
D. according to the obtained analysis of step c as a result, determining the blood biochemical markers of Wuding Chicken.
3. -2 described in any item a kind of sides based on parathormone blood biochemical markers breeding Wuding Chicken according to claim 1
Method, it is characterised in that: the blood biochemical markers of the Wuding Chicken are the concentration value of parathormone, when a monthly age, by first shape
For the concentration value of parathyrine in 167.28 pg/ml hereinafter, being determined as skeleton development normal type Wuding Chicken, on the contrary is that skeleton development is abnormal
Type Wuding Chicken；When two monthly ages, when parathormone concentration value in 172.57 pg/ml hereinafter, being determined as skeleton development normal type
Wuding Chicken, on the contrary it is skeleton development ectype Wuding Chicken.
4. -2 described in any item a kind of sides based on parathormone blood biochemical markers breeding Wuding Chicken according to claim 1
Method, it is characterised in that: cut off the water supply before blood sampling, is 12 hours jejunitas, carrying out wing venous blood sampling the next morning；Blood is collected with centrifuge tube
Sample, standing 30min, 3000rpm is centrifuged 20min at room temperature, takes supernatant, obtains serum.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811572705.5A CN109541239B (en) | 2018-12-21 | 2018-12-21 | Method for breeding Wuding chicken based on parathyroid hormone blood biochemical marker |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811572705.5A CN109541239B (en) | 2018-12-21 | 2018-12-21 | Method for breeding Wuding chicken based on parathyroid hormone blood biochemical marker |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109541239A true CN109541239A (en) | 2019-03-29 |
| CN109541239B CN109541239B (en) | 2021-09-24 |
Family
ID=65856407
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811572705.5A Active CN109541239B (en) | 2018-12-21 | 2018-12-21 | Method for breeding Wuding chicken based on parathyroid hormone blood biochemical marker |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109541239B (en) |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1123401A1 (en) * | 1998-10-22 | 2001-08-16 | The General Hospital Corporation | BIOACTIVE PEPTIDES AND PEPTIDE DERIVATIVES OF PARATHYROID HORMONE (PTH) AND PARATHYROID HORMONE-RELATED PEPTIDE (PTHrP) |
| US20080181880A1 (en) * | 2004-05-14 | 2008-07-31 | Unigene Laboratories, Inc. | Method for fostering bone formation and preservation |
| US20080213333A1 (en) * | 2004-10-25 | 2008-09-04 | Agnes Vignery | Methods and compositions for fostering and preserving bone growth |
| CN101285828A (en) * | 2008-05-30 | 2008-10-15 | 中国农业大学 | Rapid quantitative analysis method for animals skeletal muscle fiber |
| US20100048871A1 (en) * | 2004-07-21 | 2010-02-25 | Ambrx, Inc. | Biosynthetic Polypeptides Utilizing Non-Naturally Encoded Amino Acids |
| CN104650233A (en) * | 2014-03-11 | 2015-05-27 | 南京任诺药业有限公司 | ADC-drug-modified antibody IgG1-CH-K330C for establishing radioactive isotope coupling labelling |
| CN104888198A (en) * | 2015-04-21 | 2015-09-09 | 徐志强 | New application of calcitonin to preparing medicines for delaying brain aging |
| CN105028306A (en) * | 2015-06-16 | 2015-11-11 | 蚌埠市宏博生态牧业有限公司 | Novel method for breeding barred rock chicken |
| CN105510573A (en) * | 2015-12-29 | 2016-04-20 | 云南农业大学 | Method for breeding Wuding chicken by blood biochemical markers based on growth axis related genes |
| CN106938031A (en) * | 2017-03-30 | 2017-07-11 | 蒋世芬 | A kind of medicine of thyroid adenoma |
-
2018
- 2018-12-21 CN CN201811572705.5A patent/CN109541239B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1123401A1 (en) * | 1998-10-22 | 2001-08-16 | The General Hospital Corporation | BIOACTIVE PEPTIDES AND PEPTIDE DERIVATIVES OF PARATHYROID HORMONE (PTH) AND PARATHYROID HORMONE-RELATED PEPTIDE (PTHrP) |
| US20080181880A1 (en) * | 2004-05-14 | 2008-07-31 | Unigene Laboratories, Inc. | Method for fostering bone formation and preservation |
| US20100048871A1 (en) * | 2004-07-21 | 2010-02-25 | Ambrx, Inc. | Biosynthetic Polypeptides Utilizing Non-Naturally Encoded Amino Acids |
| US20080213333A1 (en) * | 2004-10-25 | 2008-09-04 | Agnes Vignery | Methods and compositions for fostering and preserving bone growth |
| CN101285828A (en) * | 2008-05-30 | 2008-10-15 | 中国农业大学 | Rapid quantitative analysis method for animals skeletal muscle fiber |
| CN104650233A (en) * | 2014-03-11 | 2015-05-27 | 南京任诺药业有限公司 | ADC-drug-modified antibody IgG1-CH-K330C for establishing radioactive isotope coupling labelling |
| CN104888198A (en) * | 2015-04-21 | 2015-09-09 | 徐志强 | New application of calcitonin to preparing medicines for delaying brain aging |
| CN105028306A (en) * | 2015-06-16 | 2015-11-11 | 蚌埠市宏博生态牧业有限公司 | Novel method for breeding barred rock chicken |
| CN105510573A (en) * | 2015-12-29 | 2016-04-20 | 云南农业大学 | Method for breeding Wuding chicken by blood biochemical markers based on growth axis related genes |
| CN106938031A (en) * | 2017-03-30 | 2017-07-11 | 蒋世芬 | A kind of medicine of thyroid adenoma |
Non-Patent Citations (4)
| Title |
|---|
| MICHAEL D ET AL.,: "Binding of Intact Parathyroid Hormone to Chicken Renal Plasma Membranes: Evidence for a Second Binding Site with Carboxyl-Terminal Specificity", 《ENDOCRINOLOGY,》 * |
| 王海鸽: "紫外线对肉仔鸡生长性能、免疫功能及骨骼发育的影响", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
| 章世元 等: "鸡甲状旁腺素基因质粒对蛋鸡蛋壳质量和激素水平的调控研究", 《浙江大学学报(农业与生命科学版)》 * |
| 豆腾飞 等: "大围山微型鸡骨代谢生化指标与骨密度研究", 《畜牧与兽医》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109541239B (en) | 2021-09-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Silverberg et al. | Skeletal disease in primary hyperparathyroidism | |
| SNOW et al. | Estrogen 2-hydroxylase oxidation and menstrual function among elite oarswomen | |
| Gracia-Marco et al. | Bone mass and bone metabolism markers during adolescence: The HELENA Study | |
| Inoue et al. | Knee osteoarthritis, knee joint pain and aging in relation to increasing serum hyaluronan level in the Japanese population | |
| Slater Jr et al. | Monoclonal antibodies that detect biochemical markers of arthritis in humans | |
| Zamboni et al. | Obesity and regional body-fat distribution in men: separate and joint relationships to glucose tolerance and plasma lipoproteins | |
| Smith et al. | The effects of acute exercise on bone turnover markers in middle-aged and older adults: A systematic review | |
| Miyamoto et al. | Collagen cross-link metabolites in urine as markers of bone metastases in prostatic carcinoma | |
| Brodin et al. | Serum levels of vitamin D are not associated with future risk of venous thromboembolism | |
| Thorland et al. | Anthropometric characteristics of elite adolescent competitive swimmers | |
| Xu et al. | Fat mass accumulation compromises bone adaptation to load in finnish women: A cross‐sectional study spanning three generations | |
| Tie et al. | Measurement of bone mineral density in the tunnel regions for anterior cruciate ligament reconstruction by dual-energy X-ray absorptiometry, computed tomography scan, and the immersion technique based on Archimedes' principle | |
| Thompson et al. | Environmental, dietary, and behavioral factors distinguish Chinese adults with high waist-to-height ratio with and without inflammation | |
| Carbone et al. | Bone density in Ehlers-Danlos syndrome | |
| Menon et al. | Expression of fibroblast growth factor 23, vitamin D receptor, and sclerostin in bone tissue from hypercalciuric stone formers | |
| Gholami et al. | Dietary acid load and bone health: a systematic review and meta-analysis of observational studies | |
| Zafeirakis et al. | Clinical and imaging correlations of bone turnover markers in prostate cancer patients with bone only metastases | |
| Reid et al. | Fiber types and fiber diameters in canine respiratory muscles | |
| Park et al. | Estrogen deficiency and low-calcium diet increased bone loss and urinary calcium excretion but did not alter arterial stiffness in young female rats | |
| CN109541239A (en) | A method of based on parathormone blood biochemical markers breeding Wuding Chicken | |
| CN109855983A (en) | A method of based on osteoprotegerin blood biochemical markers breeding Wuding Chicken | |
| CN109601477A (en) | A method of based on vitamin D receptor blood biochemical markers breeding Wuding Chicken | |
| Kim et al. | Comparative assessment of bone among wild-type, restricted ovulator and out-of-production hens | |
| Hamidpour et al. | Evaluation of relationship between biochemical parameters and osteoporosis in patients with β-thalassemia major | |
| Beyitler et al. | Impact of Mediterranean climate and seasonal variation on vitamin D levels in children |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |