CN109709339A - Detect colloidal gold immuno-chromatography test paper strip and the application of ox or ovine skeletal muscle Troponin I - Google Patents
Detect colloidal gold immuno-chromatography test paper strip and the application of ox or ovine skeletal muscle Troponin I Download PDFInfo
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Abstract
The present invention relates to a kind of detection oxen or the colloid gold chromatographic test paper strip of ovine skeletal muscle Troponin I and the preparation method and application thereof, belong to immunological technique field and Food Safety Analysis technical field.The test strips structure includes bottom plate, sample pad, colloidal gold pad, coated film and water absorption pad, and test strips both ends are equipped with protective film, and the sample pad, colloidal gold pad, coated film, water absorption pad are successively pasted on bottom plate；The capture antibody of colloid gold label is coated in colloidal gold pad；Coated film is equipped with the stealthy detection line of detection antibody-solutions printing, and the stealthy nature controlling line printed with goat anti-mouse igg solution.The capture antibody is secreted to obtain by the hybridoma cell strain 3A8 that deposit number is CCTCC NO:C2018217, and the detection antibody is secreted to obtain by the hybridoma cell strain 3D3 that deposit number is CCTCC NO:C2018218.Colloidal gold chromatographic test strip of the invention has the characteristics that convenient, quick, intuitive, timeliness is strong, applied widely, at low cost, easy to promote and utilize.
Description
Technical field
The present invention relates to colloidal gold immuno-chromatography test paper strip and its preparations of a kind of detection ox or ovine skeletal muscle Troponin I
Method and application, belong to immunological technique field and Food Safety Analysis technical field.
Background technique
In meat product, due to price, religion, health etc., it is true that many countries formulate laws and regulations requirement food labelling
It is real clearly to mark meat sources, forbid adulterated behavior, to protect the interests of consumer, but obscures meat kind in the market
Phenomenon is still very universal, and adulterated mode includes the means such as mixing, being mixed into, extracting, palming off, especially with beef or mutton product
Doping, adulterated behavior are most commonly seen, these adulterated behaviors greatly compromise the interests of consumer.
Currently, many laboratories all use the derived component in Protocols in Molecular Biology means detection meat products both at home and abroad,
DNA detection method means multiplicity, predominantly nucleic acid probe hybridization, DNA fingerprint analysis, PCR specific amplified, PCR-RFLP etc..This
A little methods have certain sensitivity, however, there are also it is at high cost, height, heavy workload, Bu Nengxian are required to test object
, there is very big uncertainty in the disadvantages of field detecting especially for cooked meat product detection, because this is largely dependent upon cold cuts
The treatment process of product, the diversity of production process result in the different Degradation Levels of genetic stew.Furthermore this method can only be examined
Survey the possibility source of animal DNA, it is necessary to the moment prevents cross contamination, milk, blood, fat be likely to be DNA source.Cause
This, needs other methods mutually to confirm come its testing result with him.Immunological method have high sensitivity, specificity it is good,
The features such as at low cost, easy to operate, is suitable for batch samples and screens, and wherein enzyme linked immunosorbent assay and colloidal gold strip are most
Common method.
However cooked meat product will generally pass through high temperature, HIGH PRESSURE TREATMENT, many albumen are all become in this process
Property, antigenicity and water solubility are lost, this just brings difficulty to the foundation of immunological method.To Immunological Method is applied to
The detection of animal derived materials, it is necessary to find a specific heat-staple protein as marker antigen, then develop
Out for the monoclonal antibody of the specificity of this antigen.Troponin I (TnI) in animals skeletal muscle has kind specificity,
It can be used as a kind of heat-resisting type marker protein, distinguish the meat type source of different genera life, cooked meat product.With ox or sheep bone bone
Flesh Troponin I (skTnI) is target detection thing, is able to achieve building for ox or mutton derived component immunology detection authentication technique
It is vertical.Therefore, the preparation of ox or ovine skeletal muscle Troponin I colloidal gold immuno-chromatography test paper strip is for carrying out beef or mutton meat source
The immunological test identification work of ingredient has important practical significance and social effect.
Summary of the invention
The technical problem to be solved by the present invention is to overcome prior art defect to provide a kind of detection ox or ovine skeletal muscle flesh
The colloidal gold immuno-chromatography test paper strip of calcium protein I, the colloidal gold immuno-chromatography test paper strip have easy to operate, quickly, are suitble to big
The advantages of batch sample primary dcreening operation.In addition, the present invention further provides the colloidal golds of the detection ox or ovine skeletal muscle Troponin I to exempt from
The preparation method and application of epidemic disease chromatograph test strip.
Technical problem of the present invention is realized by the following technical solutions.
A kind of colloidal gold immuno-chromatography test paper strip detecting ox or ovine skeletal muscle Troponin I, the test strip is the bottom of by
Plate, sample pad, colloidal gold pad, coated film and water absorption pad composition, test paper both ends are equipped with protective film；The sample pad, colloidal gold pad,
Coated film and water absorption pad are successively pasted on bottom plate；Wherein colloidal gold pad is the capture antibody for being adsorbed with colloid gold label, coating
Contain the useful pre-coated detection line T line of detection antibody, with the pre-coated nature controlling line C line of goat-anti or rabbit anti-mouse IgG solution, two on film
Bar line is arranged in parallel vertically.
Above-mentioned colloidal gold immuno-chromatography test paper strip, the capture antibody are CCTCC NO:C2018217's by deposit number
Hybridoma cell strain 3A8 secretes to obtain, the hybridoma that the detection antibody is CCTCC NO:C2018218 by deposit number
Strain 3D3 secretes to obtain；Two plants of cell strains deliver China typical culture collection center (CCTCC) guarantor on December 20th, 2018
Hiding.
Above-mentioned colloidal gold immuno-chromatography test paper strip, the capture antibody and detection antibody are source of mouse, Ma Yuan, Yang Yuan, rabbit source
Or cavy resource monoclonal antibody, preferred source of mouse monoclonal antibody；It extracts immunizing antigen by ox or ovine skeletal muscle Troponin I and exempts from
Epidemic disease obtains.
Above-mentioned colloidal gold immuno-chromatography test paper strip, the bottom plate are PVC item or other hard materials not absorbed water；The sample
Product pad and colloidal gold pad are made of glass fibre cotton；The water absorption pad is made of absorbent filter；The coated film can be fine for nitric acid
Tie up plain film or cellulose acetate film；The protective film is opaque glue film.
Colloid gold chromatographic test paper strip preparation method of the present invention, includes the following steps:
(1) the capture antibody of colloid gold label is prepared:
Capture antibody (3D3) solution 10000r/min to be marked is centrifuged 30min；Colloidal gold solution 10mL is taken, is used
0.1mol/L K2CO3Solution adjusts the pH value of colloidal gold solution to 8.2；Take equivalent to capture antibody-solutions, under magnetic agitation with glue
The mixing of body gold solution is incubated at room temperature 15min, and 10000r/min is centrifuged 30min, abandons supernatant, and gained precipitating is contained 10g with every liter
BSA, 0.5g Sodium azide, the Na that concentration is 0.02mol/L2B4O7Solution dilution, i.e. the capture antibody of acquisition colloid gold label, 4 DEG C
It saves backup；
(2) prepared by colloidal gold pad: glass fibre cotton is cut by specification, the capture antibody gold spraying instrument of colloid gold label is equal
Even to be sprayed on glass fibre cotton, 37 DEG C of dry 1h, sealing, 4 DEG C save backup；
(3) preparation of detection line and nature controlling line: it will test antibody (3A8) and be put in gold spraying instrument storage pool A, goat anti-mouse igg
Solution is put in storage pool B, and fixed fire forms the linear stealthy detection line and stealthy matter of spacing 0.5cm in film center respectively for booting
Line is controlled, is spontaneously dried, sealing, 4 DEG C save backup；
(4) test strips assemble: sample pad, colloidal gold pad, coated film and water absorption pad are successively pasted on bottom plate in order,
And the surface mount protective film at test strips both ends, test strips are cut on slot-cutting machine.
The colloid gold chromatographic test paper strip ox or ovine skeletal muscle Troponin I in for fresh meat and its product are answered
With.
The above-mentioned application for detecting ox or ovine skeletal muscle Troponin I, includes the following steps:
(1) sample pre-treatments
Fresh meat removal fat and connective tissue, meat processing food remove casing, weigh the NaCL that 0.15M is added in 1g
Solution 2ml (1:2w/v), heats 20min with boiling water after homogenate, and 2000g is centrifuged 30min；Removal precipitating, supernatant Whatman1
The filtering of number filter paper collects filtrate for detecting；
(2) it is detected with colloid gold chromatographic test paper strip
The colloidal gold immuno-chromatography test paper strip for taking out the detection ox or ovine skeletal muscle Troponin I from the package, by sample
Product end is inserted into analyte sample fluid, and insertion depth is no more than mark line, and about 10-20 seconds taking-up test strip is horizontally arranged,
Observation in 3-5 minutes determines testing result, and result is invalid after 10 minutes；
(3) result judgement
(a) it is positive: if corresponding quality control region location of C shows that a reddish brown colo(u)r streak, detection zone T location are shown on coated film
Reddish brown colo(u)r streak indicates that testing result is the positive, illustrates to contain ox or ovine skeletal muscle Troponin I in sample to be tested；
(b) negative: if T location does not develop the color on coated film, to show a reddish brown colo(u)r streak in location of C, indicate result
For feminine gender, illustrate in sample to be tested without ox or ovine skeletal muscle Troponin I；
(c) it fails: whether quality control region C does not show brownish red band, then no matter detection zone T shows brownish red band
The test paper is judged in vain.
Colloid gold chromatographic test paper strip of the invention is limited to 20mg/L to the lowest detection of ox or ovine skeletal muscle Troponin I,
Detection sensitivity to beef and mutton is 5%, is lower than 0.1% with chicken, duck, Animal muscles Troponin I extract cross reacting rate.
The method for detecting ox or ovine skeletal muscle Troponin I with colloid gold chromatographic test paper strip of the present invention is easy, quick, intuitive, accurate, suitable
With wide, at low cost, the easy popularization and use of range.
Detailed description of the invention
Fig. 1 detection ox of the present invention or ovine skeletal muscle Troponin I colloid gold chromatographic test paper strip test paper cross-section structure show
It is intended to
Fig. 2 detection ox of the present invention or ovine skeletal muscle Troponin I colloid gold chromatographic test paper strip test paper plan structure are shown
It is intended to
Fig. 3 detection ox of the present invention or ovine skeletal muscle Troponin I colloid gold chromatographic test paper strip test paper result process decision chart
Fig. 4 detection ox of the present invention or ovine skeletal muscle Troponin I colloid gold chromatographic test paper strip test paper sensitivity determination
Figure, Fig. 4 a cromogram, Fig. 4 b artwork master
The hybridoma cell strain skTnI-3A8 of anti-bovine muscle Troponin I monoclonal antibody of the present invention, in
Deliver China typical culture collection center (abbreviation CCTCC, address: Wuhan University, Chinese Typical Representative culture on December 20th, 2018
Object collection, postcode: 430072) preservation, deposit number CCTCC NO:C2018217.
The hybridoma cell strain skTnI-3D3 of anti-ovine skeletal muscle Troponin I monoclonal antibody of the present invention, in
Deliver China typical culture collection center (abbreviation CCTCC, address: Wuhan University, Chinese Typical Representative culture on December 20th, 2018
Object collection, postcode: 430072) preservation, deposit number CCTCC NO:C2018218.
Specific embodiment
Invention is further described in detail With reference to embodiment, and cited embodiment is not to the present invention
Content and protection scope constitute any restrictions.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
The preparation of the ox of the present invention of embodiment one or ovine skeletal muscle Troponin I antigen
The skeletal muscle of ox or sheep removal fat and connective tissue are taken, is ground, the NaCL for weighing 40g addition 0.15M is molten
Liquid (1:2w/v)；After further mixing, ultrasonic extraction 5min (50W, 20KHz), then with after boiling water heating 20min, 2000g is centrifuged
30min；Removal precipitating takes after half supernatant liquid filtering as treatment fluid 1.The other half supernatant 121 DEG C of high pressures 30min, 5000g
After being centrifuged 30min, with Whatman No. 1 filter paper filtering, 90% ethyl alcohol (1:3.74v/v) is added in filtrate, take mixed liquor 7000g from
Heart 20min is treatment fluid 2 after taking 37 DEG C of drying of precipitating, physiological saline to redissolve.Treatment fluid 1 and treatment fluid 2 are through SDS-PAGE electrophoresis
Identification, qualification result shows that ox and ovine skeletal muscle Troponin I immunogene and detection original have protein band in 21~22kD, with text
21~the 24kD of molecular weight for offering the skTnI of report is consistent.SkTnI electrophoretic band is ground, is made respectively with after normal saline dilution
For detection antigen and immunizing antigen.
The preparation of the ox of the present invention of embodiment two or ovine skeletal muscle Troponin I monoclonal antibody
1, animal immune: the female Balb/c mouse of 6-8 week old is immunized in the immunizing antigen of selective extraction, is spaced 2 weeks and is immunized
1 time, 3 times it is immune after docking take hematometry potency and inhibiting rate, select the optimal mouse of immune result to prepare fusion；
2, cell fusion: the splenocyte and mouse myeloma SP2/0 cell for the mouse for taking step 1 selected are merged,
It connects ELISA method measurement supernatant and chooses positive high hole, positive hole is subcloned by limiting dilution assay, is produced until establishing
The hybridoma cell strain of the monoclonal antibody of raw single anti-ox or ovine skeletal muscle Troponin I；
3, a large amount of preparations of monoclonal antibody: choosing the biggish female Balb/c mouse of individual, induces ascites using internal
Method largely prepares ascites, and purifies ascites by octanoic acid-ammonium sulfate precipitation, is divided into tubule, and -20 DEG C of preservations obtain ox or sheep bone
Bone flesh Troponin I monoclonal antibody.
The capture antibody of the present invention of embodiment three and detection antibody characteristic identification
1, titration
Will test antigen diluent with the carbonate buffer solution of pH9.6 is that 5 μ g/ml are coated with detection plate, by Dan Ke after purification
Grand antibody carries out 1:2000, and 1:4000,1:8000 ... ... 1:1024000 dilution are added in ELISA Plate hole, are added after reaction
The sheep anti mouse secondary antibody of HRP label, is finally developed the color with TMB, the results show that bovine muscle Troponin I monoclonal after purification is anti-
Potency when bulk concentration is 1mg/mL reaches 1:106, ovine skeletal muscle Troponin I MAb concentration after purification is 1mg/mL
When potency reach 1:2.56 × 105。
2, hypotype measures
Hypotype measurement is carried out using the mouse source monoclonal antibody hypotype identification kit purchased from Sigma company, the results show that ox bone bone
Flesh Troponin I monoclonal antibody hypotype is IgG1, and ovine skeletal muscle Troponin I monoclonal antibody hypotype is IgM.
3, affinity determination
Using the affinity costant of Dot-ELISA measurement monoclonal antibody, the results show that anti-bovine muscle flesh calcium egg
White I monoclonal affinity costant is Ka=8.1 × 108L/mol, anti-ovine skeletal muscle Troponin I monoclonal affinity costant are Ka=
7.6×108L/mol。
Example IV colloid gold chromatographic test paper strip of the present invention
1, prepared by colloidal gold liquid: take 100mL distilled water, boil 5min, be added gold chloride 1mL that mass concentration is 1% and
The citric acid three sodium solution 1.5mL that mass concentration is 1%, continues agitating and heating, and solution eventually passes through grey, claret to being in
Bright red is cooled to room temperature, carries out Quality Identification by ocular estimate, ultraviolet spectrophotometry respectively, qualified colloidal gold is molten
4 DEG C of liquid are kept in dark place；
2, the preparation of the capture antibody (3D3) of colloid gold label: by capture antibody-solutions 10000r/min to be marked from
Heart 30min；Colloidal gold solution 10mL is taken, with 0.1mol/L K2CO3Solution adjusts the pH value of colloidal gold solution to 8.2；Take equivalent
Antibody-solutions are captured, are mixed under magnetic agitation with colloidal gold solution, 15min is incubated at room temperature, 10000r/min is centrifuged 30min, abandons
Supernatant, the Na for being 0.02mol/L with every liter of BSA containing 10g, 0.5g Sodium azide, concentration by gained precipitating2B4O7Solution dilution, i.e.,
The capture antibody of colloid gold label is obtained, 4 DEG C save backup；
3, prepared by colloidal gold pad: cutting glass fibre cotton by specification, gold labeling antibody is uniformly sprayed on glass with gold spraying instrument
On cellucotton, 37 DEG C of dry 1h, sealing, 4 DEG C are saved backup；
4, the preparation of detection line and nature controlling line: it will test antibody (3A8) and be put in gold spraying instrument storage pool A, goat anti-mouse igg is molten
Liquid is put in storage pool B, and fixed fire is central in film respectively for booting, the linear stealthy detection line T line of formation spacing 0.5cm and stealthy matter
Line C line is controlled, is spontaneously dried, sealing, 4 DEG C save backup；
5, test strips assemble: sample pad, colloidal gold pad, coated film and water absorption pad are successively pasted on bottom plate in order,
And the surface mount protective film at test strips both ends, test strips are cut on slot-cutting machine.
6, the structure of colloid gold chromatographic test paper strip, referring to Fig. 1, Fig. 2.Figure insole board 1 is made of PVC board, and sample pad 2 uses glass
Glass cellucotton is made, and capture antibody is adsorbed in colloidal gold pad 3, and coated film 4 uses nitrocellulose filter, the water suction of water absorption pad 7
Filter paper is made, and each layer of number 2,3,4,7 is successively pasted and fixed on bottom plate 1 from right to left, intersection fiber mutually intersects each other
Fork infiltration.Stealthy detection line 5 and stealthy nature controlling line 6 are equipped on coated film 4, the detection antibody printing of stealthy detection line is stealthy
Nature controlling line is printed with goat anti-mouse igg antibody solution, and two traces are arranged in parallel at " ︱ ︱ ".8 be to be covered on sample pad 2 and colloid
Sample end protective film above gold pad 3,9 be the handle end protective film being covered on above water absorption pad 7, on the right side of sample mark line
Arrow and MAX printed words are printed on protective film.
Ox or ovine skeletal muscle Troponin I in the colloid gold chromatographic test paper strip test sample of the present invention of embodiment five
1, the pre-treatment of sample
Fresh meat removal fat and connective tissue, meat processing food remove casing, and the NaCL for weighing 1g addition 0.15M is molten
Liquid 2ml (1:2w/v), heats 20min with boiling water after homogenate, and 2000g is centrifuged 30min；Removal precipitating, supernatant is with Whatman 1
Filter paper filtering collects filtrate for detecting.
(2) it is detected with colloid gold chromatographic test paper strip
The colloidal gold immuno-chromatography test paper strip for taking out the detection ox or ovine skeletal muscle Troponin I from the package, by sample
Product end is inserted into analyte sample fluid, and insertion depth is no more than mark line, and about 10-20 seconds taking-up test strip is horizontally arranged,
Observation in 3-5 minutes determines testing result, and result is invalid after 10 minutes.
(3) result judgement
As shown in figure 3, a is feminine gender, corresponding quality control region location of C shows a reddish brown colo(u)r streak on coated film, detection zone T
It sets and does not show reddish brown colo(u)r streak, indicate that testing result is feminine gender, illustrate in sample to be tested without ox or ovine skeletal muscle troponin
I；B is the positive, and T, location of C show two days reddish brown colo(u)r streaks on coated film, indicates that result is the positive, illustrates in sample to be tested
Contain ox or ovine skeletal muscle Troponin I；C, d is failure, when quality control region C does not show brownish red band, then no matter detection zone T
The test paper is judged in vain whether showing brownish red band.
The colloid gold chromatographic test paper strip performance detection of the present invention of embodiment six
1, sensitivity
Ox or ovine skeletal muscle Troponin I are diluted to the series of standards of 30,25,20,15,10mg/L respectively with PBS
Solution, label are respectively 1,2,3,4,5, then take 90 μ L above-mentioned standard solution drop in the sample pad of test strips respectively, after 3min
Observe the colour developing situation of test strips.
As a result as shown in Fig. 2, T line starts to develop the color, and illustrates the detection of colloidal gold strip when adding concentration is 20 μ g/L
It is limited to 20mg/L.
2, specific
Chicken, duck, Animal muscles troponin extract are chosen, is diluted to 100mg/L test with PBS, observation test strips are aobvious
Color effect, testing result show that cross reaction is not present in the colloidal gold strip of preparation and chicken, duck, Animal muscles extract.
The technical concept and advantage of above-described embodiment only to illustrate the invention, the present invention also can have other forms and become
Change, as well known to the skilled person, above-described embodiment is functioned only as to the exemplary role in foregoing invention protection scope, right
For those of ordinary skill in the art, there are also many conventional deformations and other implementations in the protection scope defined by the present invention
Example, these deformations and embodiment all will be within the pending protection scopes of the present invention.
Claims (7)
1. the colloidal gold immuno-chromatography test paper strip of a kind of detection ox or ovine skeletal muscle Troponin I, which is characterized in that detection examination
Paper slip is made of bottom plate, sample pad, colloidal gold pad, coated film and water absorption pad, and test paper both ends are equipped with protective film；The sample pad,
Colloidal gold pad, coated film and water absorption pad are successively pasted on bottom plate；Wherein colloidal gold pad is to be adsorbed with the capture of colloid gold label
Antibody contains the useful pre-coated detection line T line of detection antibody on coated film, with the pre-coated matter of goat-anti or rabbit anti-mouse IgG solution
Line C line is controlled, two lines are arranged in parallel vertically.
2. colloid gold chromatographic test paper strip according to claim 1, which is characterized in that the bottom plate be PVC item or it is other not
The hard material of water suction；The sample pad and colloidal gold pad are made of glass fibre cotton；The water absorption pad is made of absorbent filter；
The coated film can be nitrocellulose filter or cellulose acetate film；The protective film is opaque glue film.
3. colloid gold chromatographic test paper strip according to claim 1, which is characterized in that the capture antibody is by deposit number
The hybridoma cell strain 3A8 of CCTCC NO:C2018217 secretes to obtain, and the detection antibody is CCTCC NO by deposit number:
The hybridoma cell strain 3D3 of C2018218 secretes to obtain；Two plants of cell strains deliver Chinese Typical Representative culture on December 20th, 2018
Object collection (CCTCC) preservation.
4. colloid gold chromatographic test paper strip according to claim 1, which is characterized in that the capture antibody and detection antibody are
Source of mouse, Ma Yuan, Yang Yuan, rabbit source or cavy resource monoclonal antibody, preferably source of mouse monoclonal antibody；It is by ox or ovine skeletal muscle flesh calcium
Protein I extraction immunizing antigen is immune to be obtained.
5. preparation detection detection ox or ovine skeletal muscle Troponin I colloidal gold immuno-chromatography test paper strip as described in claim 1
Preparation method, which comprises the steps of:
(1) the capture antibody of colloid gold label is prepared:
Capture antibody (3D3) solution 10000r/min to be marked is centrifuged 30min；Colloidal gold solution 10mL is taken, 0.1mol/ is used
L K2CO3Solution adjusts the pH value of colloidal gold solution to 8.2；Take equivalent to capture antibody-solutions, under magnetic agitation with colloidal gold solution
Mixing is incubated at room temperature 15min, and 10000r/min is centrifuged 30min, abandons supernatant, by gained precipitating every liter of BSA containing 10g, 0.5g
Sodium azide, the Na that concentration is 0.02mol/L2B4O7Solution dilution, that is, obtain the capture antibody of colloid gold label, and 4 DEG C of preservations are standby
With；
(2) prepared by colloidal gold pad: cutting glass fibre cotton by specification, the capture antibody of colloid gold label is uniformly sprayed with gold spraying instrument
It spills on glass fibre cotton, 37 DEG C of dry 1h, sealing, 4 DEG C save backup；
(3) preparation of detection line and nature controlling line: it will test antibody (3A8) and be put in gold spraying instrument storage pool A, goat anti-mouse igg solution
It is put in storage pool B, fixed fire is central in film respectively for booting, the linear stealthy detection line of formation spacing 0.5cm and stealthy nature controlling line,
It spontaneously dries, sealing, 4 DEG C save backup；
(4) test strips assemble: sample pad, colloidal gold pad, coated film and water absorption pad are successively pasted on bottom plate in order, and
The surface mount protective film at test strips both ends, is cut into test strips on slot-cutting machine.
6. colloid gold chromatographic test paper strip as described in claim 1 ox or ovine skeletal muscle flesh calcium in for fresh meat and its product
The application of protein I.
7. application according to claim 6, which comprises the steps of:
(1) sample pre-treatments
Fresh meat removal fat and connective tissue, meat processing food remove casing, weigh the NaCL solution that 0.15M is added in 1g
2ml (1:2w/v), heats 20min with boiling water after homogenate, and 2000g is centrifuged 30min；Removal precipitating, supernatant are filtered with Whatman 1
Paper filtering collects filtrate for detecting；
(2) it is detected with colloid gold chromatographic test paper strip
The colloidal gold immuno-chromatography test paper strip for taking out the detection ox or ovine skeletal muscle Troponin I from the package, by sample end
It is inserted into analyte sample fluid, insertion depth is no more than mark line, and about 10-20 seconds taking-up test strip is horizontally arranged, 3-5
Minute observation determines testing result, and result is invalid after 10 minutes；
(3) result judgement
(a) it is positive: if corresponding quality control region location of C shows that a reddish brown colo(u)r streak, detection zone T location show reddish brown on coated film
Colo(u)r streak indicates that testing result is the positive, illustrates to contain ox or ovine skeletal muscle Troponin I in sample to be tested；
(b) negative: if T location does not develop the color on coated film, to show a reddish brown colo(u)r streak in location of C, indicate that result is yin
Property, illustrate in sample to be tested without ox or ovine skeletal muscle Troponin I；
(c) it fails: examination whether quality control region C does not show brownish red band, then no matter detection zone T shows brownish red band
Paper is judged in vain.
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| CN201811623399.3A CN109709339B (en) | 2018-12-28 | 2018-12-28 | Colloidal gold immunochromatographic test strip for detecting skeletal muscle troponin I of cattle or sheep and application thereof |
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| CN201811623399.3A CN109709339B (en) | 2018-12-28 | 2018-12-28 | Colloidal gold immunochromatographic test strip for detecting skeletal muscle troponin I of cattle or sheep and application thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109709338A (en) * | 2018-12-28 | 2019-05-03 | 河北省科学院生物研究所 | Detect ox or the enzyme linked immunological kit of ovine skeletal muscle Troponin I and the preparation method and application thereof |
| CN109705216A (en) * | 2018-12-28 | 2019-05-03 | 河北省科学院生物研究所 | A kind of anti-bovine muscle Troponin I monoclonal antibody and its application |
| CN109810191A (en) * | 2018-12-28 | 2019-05-28 | 河北省科学院生物研究所 | A kind of anti-ovine skeletal muscle Troponin I monoclonal antibody and its application |
| CN111220809A (en) * | 2019-12-27 | 2020-06-02 | 河北省科学院生物研究所 | Colloidal gold immunochromatographic test strip for detecting chicken or duck skeletal muscle troponin I and preparation method and application thereof |
| CN113215274A (en) * | 2021-05-10 | 2021-08-06 | 中国农业科学院农业质量标准与检测技术研究所 | miRNA detection method and lateral flow chromatography test strip for detecting miRNA |
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| WO2004076684A2 (en) * | 2003-02-27 | 2004-09-10 | Strategic Diagnostics Inc. | Compositions and methods for the detection of mammalian muscle troponin i |
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| CN109709338A (en) * | 2018-12-28 | 2019-05-03 | 河北省科学院生物研究所 | Detect ox or the enzyme linked immunological kit of ovine skeletal muscle Troponin I and the preparation method and application thereof |
| CN109705216A (en) * | 2018-12-28 | 2019-05-03 | 河北省科学院生物研究所 | A kind of anti-bovine muscle Troponin I monoclonal antibody and its application |
| CN109810191A (en) * | 2018-12-28 | 2019-05-28 | 河北省科学院生物研究所 | A kind of anti-ovine skeletal muscle Troponin I monoclonal antibody and its application |
| CN109810191B (en) * | 2018-12-28 | 2021-09-07 | 河北省科学院生物研究所 | Monoclonal antibody for resisting sheep skeletal muscle troponin I and application thereof |
| CN109705216B (en) * | 2018-12-28 | 2021-09-07 | 河北省科学院生物研究所 | Monoclonal antibody for resisting bovine skeletal muscle troponin I and application thereof |
| CN109709338B (en) * | 2018-12-28 | 2022-03-15 | 河北省科学院生物研究所 | Enzyme linked immunosorbent assay kit for detecting skeletal muscle troponin I of cattle or sheep and preparation method and application thereof |
| CN111220809A (en) * | 2019-12-27 | 2020-06-02 | 河北省科学院生物研究所 | Colloidal gold immunochromatographic test strip for detecting chicken or duck skeletal muscle troponin I and preparation method and application thereof |
| CN111220809B (en) * | 2019-12-27 | 2023-06-16 | 河北省科学院生物研究所 | Colloidal gold immunochromatography test strip for detecting chicken or duck skeletal muscle troponin I and preparation method and application thereof |
| CN113215274A (en) * | 2021-05-10 | 2021-08-06 | 中国农业科学院农业质量标准与检测技术研究所 | miRNA detection method and lateral flow chromatography test strip for detecting miRNA |
| CN113215274B (en) * | 2021-05-10 | 2022-07-29 | 中国农业科学院农业质量标准与检测技术研究所 | miRNA detection method and lateral flow chromatography test strip for detecting miRNA |
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