CN109709339B - Colloidal gold immunochromatographic test strip for detecting skeletal muscle troponin I of cattle or sheep and application thereof - Google Patents
Colloidal gold immunochromatographic test strip for detecting skeletal muscle troponin I of cattle or sheep and application thereof Download PDFInfo
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Abstract
The invention relates to a colloidal gold chromatography test strip for detecting skeletal muscle troponin I of cattle or sheep, a preparation method and application thereof, belonging to the technical field of immunology and the technical field of food safety analysis. The test strip structure comprises a bottom plate, a sample pad, a colloidal gold pad, a coating film and a water absorption pad, wherein protective films are arranged at two ends of the test strip, and the sample pad, the colloidal gold pad, the coating film and the water absorption pad are sequentially adhered to the bottom plate; the colloidal gold pad is coated with a capture antibody marked by colloidal gold; the envelope film is provided with a hidden detection line printed by a detection antibody solution and a hidden quality control line printed by a goat anti-mouse IgG solution. The capture antibody is obtained by secreting a hybridoma cell strain skTnI-3A8 with the preservation number of CCTCC NO: C2018217, and the detection antibody is obtained by secreting a hybridoma cell strain skTnI-3D3 with the preservation number of CCTCC NO: C2018218. The colloidal gold chromatography detection test strip has the characteristics of convenience, rapidness, intuition, strong timeliness and the like, and is wide in application range, low in cost and convenient to popularize and apply.
Description
Technical Field
The invention relates to a colloidal gold immunochromatographic test strip for detecting skeletal muscle troponin I of cattle or sheep, a preparation method and application thereof, belonging to the technical field of immunology and the technical field of food safety analysis.
Background
In meat products, due to reasons of price, religion, health and the like, regulations established by many countries require food labels to truly and definitely mark meat sources and prohibit adulteration behaviors so as to protect the benefits of consumers, but phenomena of confusing meat varieties in the market are still very common, adulteration modes comprise means of adulteration, mixing, extraction, counterfeiting and the like, particularly the most common behaviors of adulteration and adulteration of beef and mutton products, and the adulteration behaviors greatly damage the benefits of the consumers.
At present, many laboratories at home and abroad use molecular biology technical means to detect source components in meat products, and DNA detection methods are various and mainly comprise nucleic acid probe hybridization, DNA fingerprint analysis, PCR specific amplification, PCR-RFLP and the like. These methods have a certain sensitivity, but also have the disadvantages of high cost, high requirement on detection objects, large workload, incapability of on-site detection and the like, and particularly have great uncertainty on detection of cooked meat products, because the methods depend on the processing process of the cooked meat products to a great extent, and the diversity of the production process causes different degradation levels of gene substances. Furthermore, this method only detects possible sources of animal DNA, and must be done at all times to prevent cross-contamination, and milk, blood, and fat are all possible sources of DNA. Therefore, other methods are needed to mutually corroborate with other test results. The immunological method has the characteristics of high sensitivity, good specificity, low cost, convenient operation and the like, and is suitable for screening large-batch samples, wherein an enzyme-linked immunosorbent assay and a colloidal gold test strip are the most common methods.
However, cooked meat products are generally subjected to high temperature and high pressure treatment, and many proteins are denatured in the process, so that antigenicity and water solubility are lost, and the establishment of an immunological method is difficult. In order to apply the immunological method to the detection of animal-derived components, a specific thermostable protein must be found as a marker antigen, and then a specific monoclonal antibody against the antigen must be developed. Troponin I (TnI) in animal skeletal muscle has species specificity, and can be used as a heat-resistant species marker protein for distinguishing meat species sources of different species of raw and cooked meat products. The bovine or sheep skeletal muscle troponin I (skTnI) is taken as a target detection object, and the establishment of the immunological detection and identification technology of bovine or sheep meat-derived components can be realized. Therefore, the preparation of the colloidal gold immunochromatographic test strip for skeletal muscle troponin I of cattle or sheep has important practical significance and social significance for carrying out the immunological test and identification work of beef or mutton meat components.
Disclosure of Invention
The invention aims to solve the technical problem of overcoming the defects of the prior art and provides the colloidal gold immunochromatographic test strip for detecting the skeletal muscle troponin I of the cattle or sheep, and the colloidal gold immunochromatographic test strip has the advantages of simple and quick operation and suitability for preliminary screening of large-batch samples. In addition, the invention further provides a preparation method and application of the colloidal gold immunochromatographic test strip for detecting skeletal muscle troponin I of cattle or sheep.
The technical problem of the invention is realized by the following technical scheme.
A colloidal gold immunochromatographic test strip for detecting skeletal muscle troponin I of cattle or sheep comprises a bottom plate, a sample pad, a colloidal gold pad, a coating film and a water absorption pad, wherein protective films are arranged at two ends of the test strip; the sample pad, the colloidal gold pad, the coating film and the water absorption pad are sequentially adhered to the bottom plate; the colloidal gold pad is a capture antibody adsorbed with a colloidal gold label, the coating film contains a detection line T line pre-coated with the detection antibody and a quality control line C line pre-coated with goat anti or rabbit anti-mouse IgG solution, and the two lines are arranged vertically and parallelly.
The capture antibody is secreted by a hybridoma cell strain skTnI-3A8 with the preservation number of CCTCC NO of C2018217, and the detection antibody is secreted by a hybridoma cell strain skTnI-3D3 with the preservation number of CCTCC NO of C2018218; the two cell strains are delivered to China Center for Type Culture Collection (CCTCC) for preservation in 2018, 12 months and 20 days.
According to the colloidal gold immunochromatographic test strip, the capture antibody and the detection antibody are murine, equine, ovine, rabbit or guinea pig monoclonal antibodies, preferably murine monoclonal antibodies; it is obtained by immunizing bovine or ovine skeletal muscle troponin I with an immunizing antigen.
In the colloidal gold immunochromatographic test strip, the base plate is a PVC strip or other hard materials which do not absorb water; the sample pad and the colloidal gold pad are made of glass fiber cotton; the water absorption pad is made of water absorption filter paper; the coating film can be a nitrocellulose film or a cellulose acetate film; the protective film is an opaque adhesive film.
The preparation method of the colloidal gold chromatography test strip comprises the following steps:
(1) preparation of colloidal gold labeled capture antibody:
will be markedThe solution of the capture antibody skTnI-3A8 is centrifuged at 10000r/min for 30 min; taking 10mL of colloidal gold solution, and using 0.1mol/L K2CO3Adjusting the pH value of the colloidal gold solution to 8.2 by using the solution; collecting equal amount of capture antibody solution, mixing with colloidal gold solution under magnetic stirring, incubating at room temperature for 15min, centrifuging at 10000r/min for 30min, discarding supernatant, and collecting precipitate with Na containing 10g BSA, 0.5g sodium azide and 0.02mol/L concentration2B4O7Diluting the solution to obtain a capture antibody marked by the colloidal gold, and storing the capture antibody at 4 ℃ for later use;
(2) preparing a colloidal gold pad: cutting glass fiber cotton according to specification, uniformly spraying capture antibody marked with colloidal gold on the glass fiber cotton by using a gold spraying instrument, drying at 37 ℃ for 1h, sealing, and storing at 4 ℃ for later use;
(3) preparation of detection line and quality control line: placing detection antibody skTnI-3D3 in a storage pool A of a metal spraying instrument, placing goat anti-mouse IgG solution in a storage pool B, starting the machine, respectively shooting at the center of the membrane to form a linear invisible detection line and an invisible quality control line with a distance of 0.5cm, naturally drying, sealing, and storing at 4 ℃ for later use;
(4) assembling the test strip: and sequentially adhering the sample pad, the colloidal gold pad, the coating film and the water absorption pad on the bottom plate, adhering protective films on the surfaces of two ends of the test strip, and cutting the test strip on a grooving machine.
The colloidal gold chromatography test strip is applied to the application of the skeletal muscle troponin I of cattle or sheep in fresh meat and products thereof.
The application of the protein in detecting skeletal muscle troponin I of cattle or sheep comprises the following steps:
(1) sample pretreatment
Removing fat and connective tissue from fresh meat, processing meat food, removing casing, weighing 1g, adding 0.15M NaCl solution 2ml (1:2w/v), homogenizing, heating with boiling water for 20min, and centrifuging at 2000g for 30 min; removing precipitate, filtering supernatant with Whatman No. 1 filter paper, and collecting filtrate for detection;
(2) detection with colloidal gold chromatography test paper strip
Taking out the colloidal gold immunochromatographic test strip for detecting the skeletal muscle troponin I of the cattle or sheep from the package, inserting the sample end into the sample solution to be detected, wherein the insertion depth does not exceed the mark line, taking out the test strip in about 10-20 seconds, horizontally placing, observing for 3-5 minutes to judge the detection result, and after 10 minutes, the result is invalid;
(3) determination of results
(a) Positive: if a brownish red line is displayed at the position of the corresponding quality control area C on the coating film, and a brownish red line is displayed at the position of the detection area T, the detection result is positive, and the result shows that the sample to be detected contains the bovine or sheep skeletal muscle troponin I;
(b) negative: if the T position on the envelope membrane does not develop color, a brown red line is displayed on the C position, the result is negative, and the result indicates that the sample to be detected does not contain the bovine or sheep skeletal troponin I;
(c) and (3) failure: and when the quality control area C does not show a brownish red strip, the test paper is judged to be invalid whether the detection area T shows a brownish red strip or not.
The colloidal gold chromatography test strip has the lowest detection limit of 20mg/L on skeletal muscle troponin I of cattle or sheep, has the detection sensitivity of 5 percent on cattle and mutton, and has the cross reaction rate of less than 0.1 percent with chicken, duck and pig skeletal muscle troponin I extracts. The method for detecting the skeletal muscle troponin I of the cattle or sheep by using the colloidal gold chromatography test strip is simple, convenient, rapid, visual, accurate, wide in application range, low in cost and easy to popularize and use.
Drawings
FIG. 1 is a schematic diagram of a cross-sectional structure of colloidal gold chromatography test paper strip for detecting skeletal muscle troponin I of cattle or sheep
FIG. 2 is a schematic view of a top view of a colloidal gold chromatography test strip for detecting skeletal muscle troponin I of cattle or sheep
FIG. 3 is a result determination chart of colloidal gold chromatography test paper strip for detecting skeletal muscle troponin I of cattle or sheep
FIG. 4 is a graph showing the sensitivity of the colloidal gold chromatographic test strip for detecting skeletal muscle troponin I in cattle or sheep, and the graph is shown in FIG. 4a in color and FIG. 4b in black and white
The hybridoma cell strain skTnI-3A8 of the monoclonal antibody for resisting the bovine skeletal muscle troponin I is delivered to the China center for type culture Collection (CCTCC for short, address: Wuhan university, China center for type culture Collection, postal code: 430072) for preservation in 2018, 12 months and 20 days, and the preservation number is CCTCC NO: C2018217.
The hybridoma cell strain skTnI-3D3 of the monoclonal antibody for resisting the sheep skeletal muscle troponin I is delivered to a China center for type culture Collection (CCTCC for short, address: Wuhan university, China center for type culture Collection, postal code: 430072) for preservation in 2018, 12 months and 20 days, and the preservation number is CCTCC NO: C2018218.
Detailed Description
The present invention is further described in detail with reference to the following specific embodiments, which are not intended to limit the scope of the present invention.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
EXAMPLE A preparation of the skeletal muscle troponin I antigen of cattle or sheep according to the present invention
Taking cattle or sheep skeletal muscle to remove fat and connective tissue, grinding and mixing evenly, weighing 40g, adding 0.15M NaCL solution (1:2 w/v); further mixing, ultrasonic extracting for 5min (50W, 20KHz), heating with boiling water for 20min, and centrifuging at 2000g for 30 min; removing the precipitate, taking half of the supernatant, and filtering to obtain a treatment solution 1. Centrifuging the other half of the supernatant at 121 deg.C under high pressure for 30min, 5000g for 30min, filtering with Whatman No. 1 filter paper, adding 90% ethanol (1:3.74v/v) into the filtrate, centrifuging 7000g of the mixture for 20min, oven drying the precipitate at 37 deg.C, and dissolving in normal saline to obtain treated solution 2. The processing liquid 1 and the processing liquid 2 are identified by SDS-PAGE electrophoresis, and the identification result shows that the bovine and ovine skeletal muscle troponin I immunogen and the detection antigen have protein bands in the range of 21-22 kD, and the molecular weight of the protein bands is consistent with the molecular weight of 21-24 kD of skTnI reported in the literature. And grinding the sktnI electrophoresis strip, and diluting the ground sktnI electrophoresis strip with physiological saline to be respectively used as a detection antigen and an immune antigen.
EXAMPLE two preparation of monoclonal antibody against skeletal muscle troponin I of bovine or ovine of the present invention
1. Animal immunization: selecting extracted immune antigen, immunizing female Balb/c mice of 6-8 weeks old, immunizing for 1 time at intervals of 2 weeks, after 3 times of immunization, cutting tail and taking blood to measure titer and inhibition rate, and selecting the mice with the best immune result for fusion;
2. cell fusion: fusing splenocytes of the mouse selected in the step 1 with myeloma SP2/0 cells of the mouse, determining supernatant fluid by an indirect ELISA method, selecting holes with high positive, and subcloning the positive holes by a limiting dilution method until a hybridoma cell strain generating a single monoclonal antibody against bovine or sheep skeletal muscle troponin I is established;
3. large-scale preparation of monoclonal antibodies: selecting a female Balb/c mouse with a larger individual, preparing a large amount of ascites by an in vivo induced ascites method, purifying the ascites by caprylic acid-ammonium sulfate precipitation, dividing the ascites into small tubes, and storing the small tubes at the temperature of minus 20 ℃ to obtain the bovine or sheep skeletal muscle troponin I monoclonal antibody.
EXAMPLE III characterization of the Capture and detection antibodies of the invention
1. Potency assay
Diluting the detection antigen to 5 mu g/mL by using a carbonate buffer solution with pH9.6 to coat a detection plate, diluting the purified monoclonal antibody by 1:2000, 1:4000, 1:8000, … … 1:1024000, adding the diluted monoclonal antibody into an enzyme label plate hole, adding a goat-anti-mouse secondary antibody marked by HRP after reaction, and finally developing by using TMB (Tetramethylbenzidine), wherein the result shows that the titer of the purified bovine skeletal muscle troponin I monoclonal antibody reaches 1 when the concentration is 1 mg/mL: 106The titer of the purified sheep skeletal muscle troponin I monoclonal antibody reaches 1:2.56 multiplied by 10 when the concentration is 1mg/mL5。
2. Subtype determination
Subtype determination was performed using a murine monoclonal antibody subtype identification kit purchased from Sigma, and the results showed that the bovine skeletal muscle troponin I monoclonal antibody subtype was IgG1 and the ovine skeletal muscle troponin I monoclonal antibody subtype was IgM.
3. Affinity assay
The result of measuring the affinity constant of the monoclonal antibody by indirect enzyme-linked immunoassay shows that the monoclonal affinity constant of anti-bovine skeletal muscle troponin I is Ka-8.1 multiplied by 108L/mol, monoclonal anti-sheep skeletal muscle troponin IAffinity constant Ka 7.6X 108L/mol。
Example four the colloidal gold chromatography test strip of the invention
1. Preparing colloidal gold liquid: boiling 100mL of distilled water for 5min, adding 1mL of chloroauric acid with the mass concentration of 1% and 1.5mL of trisodium citrate solution with the mass concentration of 1%, continuously stirring and heating, finally carrying out gray and wine red treatment on the solution until the solution is bright red, cooling the solution to room temperature, respectively carrying out quality identification by a visual method and an ultraviolet spectrophotometry, and storing the qualified colloidal gold solution at the temperature of 4 ℃ in a dark place;
2. preparation of colloidal gold-labeled capture antibody skTnI-3 A8: centrifuging 10000r/min of capture antibody solution to be marked for 30 min; taking 10mL of colloidal gold solution, and using 0.1mol/L K2CO3Adjusting the pH value of the colloidal gold solution to 8.2 by the solution; collecting equal amount of capture antibody solution, mixing with colloidal gold solution under magnetic stirring, incubating at room temperature for 15min, centrifuging at 10000r/min for 30min, discarding supernatant, and collecting precipitate with Na containing 10g BSA, 0.5g sodium azide and 0.02mol/L concentration2B4O7Diluting the solution to obtain a capture antibody marked by the colloidal gold, and storing the capture antibody at 4 ℃ for later use;
3. preparing a colloidal gold pad: cutting glass fiber cotton according to specification, uniformly spraying gold-labeled antibody on the glass fiber cotton by using a gold spraying instrument, drying at 37 ℃ for 1h, sealing, and storing at 4 ℃ for later use;
4. preparation of detection line and quality control line: placing the detection antibody skTnI-3D3 in a storage pool A of a metal spraying instrument, placing goat anti-mouse IgG solution in a storage pool B, starting the machine, respectively shooting at the center of the membrane to form a linear invisible detection line T line and an invisible quality control line C line which are spaced by 0.5cm, naturally drying, sealing, and storing at 4 ℃ for later use;
5. assembling the test strip: sequentially adhering the sample pad, the colloidal gold pad, the coating film and the water absorption pad on the bottom plate, adhering the protective films on the surfaces of two ends of the test strip, and cutting the test strip on a grooving machine.
6. The structure of the colloidal gold chromatography test strip is shown in figure 1 and figure 2. In the figure, a bottom plate 1 is made of a PVC plate, a sample pad 2 is made of glass fiber cotton, a capture antibody is adsorbed on a colloidal gold pad 3, a coating film 4 adopts a nitrocellulose membrane, a water absorption pad 7 is made of water absorption filter paper, all layers of numbers 2, 3, 4 and 7 are sequentially stuck and fixed on the bottom plate 1 from right to left, and fibers at the junctions of the numbers are mutually crossed and permeated. An invisible detection line 5 and an invisible quality control line 6 are arranged on the coating film 4, the invisible detection line is printed by using a detection antibody, the invisible quality control line is printed by using a goat anti-mouse IgG antibody solution, and two imprints are arranged in parallel to form a 'stick'. The sample end protective film 8 is covered on the sample pad 2 and the colloidal gold pad 3, the handle end protective film 9 is covered on the water absorption pad 7, and the arrow and MAX characters are printed on the protective film on the right side of the sample marking line.
Example five the colloidal gold chromatography test strip of the invention detects skeletal muscle troponin I in cattle or sheep in a sample
1. Pretreatment of samples
Removing fat and connective tissue from fresh meat, processing meat food, removing casing, weighing 1g, adding 0.15M NaCl solution 2ml (1:2w/v), homogenizing, heating with boiling water for 20min, and centrifuging at 2000g for 30 min; the precipitate was removed, the supernatant was filtered through Whatman No. 1 filter paper, and the filtrate was collected for detection.
(2) Detection with colloidal gold chromatography test paper strip
And taking out the colloidal gold immunochromatographic test strip for detecting the skeletal muscle troponin I of the cattle or sheep from the package, inserting the sample end into the sample solution to be detected, wherein the insertion depth does not exceed the marking line, taking out the test strip in about 10-20 seconds, horizontally placing, observing for 3-5 minutes to judge the detection result, and after 10 minutes, obtaining invalid results.
(3) Determination of results
As shown in fig. 3, a is negative, a brownish red line is displayed at the position of the quality control area C corresponding to the coating film, and a brownish red line is not displayed at the position of the detection area T, which indicates that the detection result is negative, and indicates that the sample to be detected does not contain bovine or sheep skeletal troponin I; b is positive, a brown-red line of two days is displayed at the position T, C on the coating film, the result is positive, and the result indicates that the sample to be detected contains the bovine or sheep skeletal muscle troponin I; c. d is invalid, when the quality control area C does not show a brownish red strip, the test paper is judged to be invalid whether the detection area T shows a brownish red strip or not.
Sixth embodiment of the invention the colloidal gold chromatography test strip performance detection
1. Sensitivity of the probe
Diluting the skeletal muscle troponin I of cattle or sheep with PBS to obtain a series of standard solutions with the numbers of 1, 2, 3, 4 and 5 of 30, 25, 20, 15 and 10mg/L respectively, dripping 90 mu L of the standard solutions on a sample pad of a test strip, and observing the color development condition of the test strip after 3 min.
As shown in FIG. 4, when the concentration of the reagent was 20mg/L, the T-line began to develop color, indicating that the detection limit of the colloidal gold test strip was 20 mg/L.
2. Specificity of
The chicken, duck and pig skeletal muscle troponin extracts are selected, diluted to 100mg/L by PBS for testing, the color development effect of the test strip is observed, and the test result shows that the prepared colloidal gold test strip has no cross reaction with the chicken, duck and pig skeletal muscle extracts.
The above-described embodiments are intended to illustrate the technical idea and advantages of the invention, and the invention may also be subject to other variants, as known to the skilled person, which serve merely as illustrations of the scope of protection of the invention described above, and to the skilled person in the art who is within the scope of protection of the invention defined by the present invention there are many conventional variants and other embodiments, which are all within the scope of protection of the invention covered by the present invention.
Claims (4)
1. A colloidal gold immunochromatographic test strip for detecting skeletal muscle troponin I of cattle or sheep comprises a bottom plate, a sample pad, a colloidal gold pad, a coating film and a water absorption pad, wherein protective films are arranged at two ends of the test strip; the sample pad, the colloidal gold pad, the coating film and the water absorption pad are sequentially adhered to the bottom plate; wherein the colloidal gold pad is a capture antibody adsorbed with a colloidal gold label, the coating film contains a detection line T line pre-coated with the detection antibody and a quality control line C line pre-coated with goat anti or rabbit anti-mouse IgG solution, and the two lines are arranged vertically and in parallel; the bottom plate is a PVC strip or other hard materials which do not absorb water; the sample pad and the colloidal gold pad are made of glass fiber cotton; the water absorption pad is made of water absorption filter paper; the coating film is a nitrocellulose film or a cellulose acetate film; the protective film is an opaque adhesive film; the method is characterized in that the capture antibody is obtained by secreting a hybridoma cell strain skTnI-3A8 with the preservation number of CCTCC NO: C2018217, and the detection antibody is obtained by secreting a hybridoma cell strain skTnI-3D3 with the preservation number of CCTCC NO: C2018218; the two cell strains are delivered to China Center for Type Culture Collection (CCTCC) for preservation in 2018, 12 months and 20 days.
2. The method for preparing the colloidal gold immunochromatographic test strip for detecting skeletal muscle troponin I of cattle or sheep as defined in claim 1, which comprises the steps of:
(1) preparation of colloidal gold labeled capture antibody:
centrifuging 10000r/min of a solution of a capture antibody skTnI-3A8 to be marked for 30 min; taking 10mL of colloidal gold solution, and using 0.1mol/L K2CO3Adjusting the pH value of the colloidal gold solution to 8.2 by using the solution; collecting equal amount of capture antibody solution, mixing with colloidal gold solution under magnetic stirring, incubating at room temperature for 15min, centrifuging at 10000r/min for 30min, discarding supernatant, and collecting precipitate with Na containing 10g BSA, 0.5g sodium azide and 0.02mol/L2B4O7Diluting the solution to obtain a capture antibody marked by colloidal gold, and storing at 4 ℃ for later use;
(2) preparing a colloidal gold pad: cutting glass fiber cotton according to specification, uniformly spraying capture antibody marked by colloidal gold on the glass fiber cotton by a gold spraying instrument, drying at 37 ℃ for 1h, sealing, and storing at 4 ℃ for later use;
(3) preparation of detection line and quality control line: placing detection antibody skTnI-3D3 in a storage pool A of a metal spraying instrument, placing goat anti-mouse IgG solution in a storage pool B, starting the machine, respectively shooting at the center of the membrane to form a linear invisible detection line and an invisible quality control line with a distance of 0.5cm, naturally drying, sealing, and storing at 4 ℃ for later use;
(4) assembling the test strip: and sequentially adhering the sample pad, the colloidal gold pad, the coating film and the water absorption pad on the bottom plate, adhering protective films on the surfaces of two ends of the test strip, and cutting the test strip on a grooving machine.
3. The use of the colloidal gold immunochromatographic test strip of claim 1 for detecting bovine or ovine skeletal muscle troponin I in fresh meat and products thereof.
4. Use according to claim 3, characterized in that it comprises the following steps:
(1) sample pretreatment
Removing fat and connective tissue from fresh meat, processing meat food, removing casing, weighing 1g, adding 0.15M NaCl solution 2ml, homogenizing, heating with boiling water for 20min, centrifuging at 2000g for 30 min; removing precipitate, filtering the supernatant with Whatman No. 1 filter paper, and collecting filtrate for detection;
(2) detection with colloidal gold immunochromatographic test strip
Taking out the colloidal gold immunochromatographic test strip for detecting the skeletal muscle troponin I of the cattle or sheep from the package, inserting the sample end into a sample solution to be detected, wherein the insertion depth does not exceed a marking line, taking out the test strip in about 10-20 seconds, horizontally placing, observing for 3-5 minutes to judge a detection result, and after 10 minutes, obtaining an invalid result;
(3) determination of results
(a) Positive: if a brownish red line is displayed at the position of the corresponding quality control area C on the coating film, and a brownish red line is displayed at the position of the detection area T, the detection result is positive, and the result shows that the sample to be detected contains the bovine or sheep skeletal muscle troponin I;
(b) negative: if the T position on the envelope membrane does not develop color, a brown red line is displayed on the C position, the result is negative, and the result indicates that the sample to be detected does not contain the bovine or sheep skeletal troponin I;
(c) and (3) failure: and when the quality control area C does not show a brownish red strip, the test paper is judged to be invalid whether the detection area T shows a brownish red strip or not.
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| 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 |
| 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 |
| 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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