CN110554193A - Preparation method of colloidal gold immunochromatographic test strip for paraquat in urine - Google Patents
Preparation method of colloidal gold immunochromatographic test strip for paraquat in urine Download PDFInfo
- Publication number
- CN110554193A CN110554193A CN201910942583.2A CN201910942583A CN110554193A CN 110554193 A CN110554193 A CN 110554193A CN 201910942583 A CN201910942583 A CN 201910942583A CN 110554193 A CN110554193 A CN 110554193A
- Authority
- CN
- China
- Prior art keywords
- colloidal gold
- paraquat
- solution
- test strip
- urine
- 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.)
- Pending
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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
Landscapes
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to the technical field of biological detection, and particularly discloses a preparation method of a colloidal gold immunochromatographic test strip for paraquat in urine, which comprises the following steps: preparing a colloidal gold solution containing colloidal gold particles with the particle size of 10-30nm by taking a chloroauric acid aqueous solution and a trisodium citrate aqueous solution as raw materials; labeling paraquat monoclonal antibody with colloidal gold solution: centrifuging the colloidal gold solution obtained in the step a, then removing supernatant, adding the paraquat monoclonal antibody into the colloidal gold solution after the precipitate is resuspended, and standing after uniformly stirring; dropwise adding sterile bovine serum albumin, stirring, and standing overnight at room temperature; centrifuging the labeled colloidal gold solution again, discarding the supernatant, and re-dissolving the precipitate with a heavy suspension to obtain a colloidal gold-labeled paraquat monoclonal antibody solution; and then, carrying out gold spraying, coating the C line, the T line and assembling the colloidal gold immune test strip. The method is based on the colloidal gold immunochromatographic principle, and can simply, accurately and quickly screen the paraquat in the urine.
Description
Technical Field
The invention relates to the field of biological detection, in particular to a preparation method of a colloidal gold immunochromatographic test strip for paraquat in urine.
background
Paraquat, the chemical name of which is 1-1-dimethyl-4-4-bipyridine cation salt, is a quick biocidal herbicide and has a contact action and a certain systemic action. Paraquat is the most used non-selective contact herbicide in the world at present, and has the advantages of good activity, wide weed control spectrum, good effect, quick response and the like. However, paraquat has great toxicity to human, has no specific antidote, has the death rate of more than 90 percent after oral poisoning, and is a herbicide with the highest death rate of acute poisoning of human beings. Paraquat poisoning has become the pesticide poisoning type with the widest influence range and the most serious harm in the global scope, the high fatality rate of pesticide poisoning in China is the first, and the serious harm to the health of people and the harmonious and stable society are achieved.
At present, two modes of Liquid Chromatography (LC) and liquid-mass spectrometry (LC-MS) are mainly adopted for the detection method of paraquat clinically, and although the detection method of paraquat can qualitatively and quantitatively detect paraquat by means of a large instrument, the pretreatment of a sample is complex, the detection time is long, and the cost is high, so that a quick, simple, accurate and low-cost detection method is urgently needed in clinic.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, and provides a paraquat colloidal gold immune test strip in urine based on a colloidal gold immunochromatographic principle, which can simply, accurately and quickly screen paraquat in urine.
In order to achieve the purpose, the technical scheme of the invention is as follows: the preparation method of the colloidal gold immunochromatographic test strip for paraquat in urine comprises the following steps:
Step a: preparing a colloidal gold solution: preparing a colloidal gold solution containing colloidal gold particles with the particle size of 10-30nm by taking a chloroauric acid aqueous solution and a trisodium citrate aqueous solution as raw materials;
Step b: labeling paraquat monoclonal antibody with colloidal gold solution: centrifuging the colloidal gold solution in the step a, then removing supernatant, and resuspending the precipitate by using potassium carbonate solution with the pH of 8.0-8.5; adding 0.04-0.1mg of paraquat monoclonal antibody into the colloidal gold solution, stirring uniformly and standing; dropwise adding sterile bovine serum albumin with the mass concentration of 8-12%, stirring, standing overnight at room temperature to obtain a labeled colloidal gold solution; centrifuging the labeled colloidal gold solution again, discarding the supernatant, and re-dissolving the precipitate with 4-6mL of heavy suspension to obtain a colloidal gold-labeled paraquat monoclonal antibody solution;
Step c: spraying gold;
Step d: coating the nitrocellulose membrane with a C line and a T line;
Step e: and (5) assembling the colloidal gold immune test strip.
The technical scheme also provides a use method of the colloidal gold immunochromatographic test strip for paraquat in urine, which comprises the following steps:
Step (1): taking 1-3ml of biological specimen urine, adding ammonia water, adjusting the pH value to 7.0-8.0, and standing to obtain a sample solution to be detected;
step (2): dropwise adding the sample solution to be detected on a paraquat colloidal gold immunochromatographic rapid detection test strip, and timing;
and (3): reading the result after timing;
And (4): and (5) judging a result: the C line of the quality control line is displayed as a red line, when the T line is developed, the result is negative, and the concentration of paraquat in the sample to be detected is less than 50 ng/ml; when the T line does not develop color, the result is positive, the concentration of paraquat in the sample to be detected is more than 50ng/ml, the quality control line C line does not develop color, and the test strip judges that the test strip is invalid.
The principle and the beneficial effects of the technical scheme are as follows: in the technical scheme, the defect that detection of paraquat only depends on a large-scale detection instrument in the prior art is overcome, and the principle of immunochromatography is creatively adopted, but the immunochromatography technology in the prior art has the problems of low detection sensitivity and low detection precision, and the detection requirement of paraquat in urine in the technical scheme cannot be met. The inventor finds that the particle size of colloidal gold particles in a colloidal gold solution is an important factor influencing the precision of the prepared test strip in long-term research, and in the preparation process, the particle size of the colloidal gold particles in the colloidal gold solution is uniform and a dispersion system is uniform by adjusting the particle size of the colloidal gold in the colloidal gold solution to be between 10 and 30 nm; in addition, according to the technical scheme, before the potassium carbonate solution is added to adjust the pH value, the colloidal gold solution is centrifuged to remove the supernatant, and then the precipitate is adjusted by the potassium carbonate, so that the problem that the pH value of the colloidal gold solution is directly adjusted by the potassium carbonate solution in the prior art, the probe of a pH meter is easily damaged, the accuracy of the finally adjusted pH value is influenced, the agglomeration problem of the colloidal gold solution is easily caused, a stable colloidal gold solution is provided for the marking of a subsequent paraquat monoclonal antibody, and the monoclonal antibody can be stably adsorbed and combined on the surface of a colloidal gold particle.
According to the technical scheme, the whole preparation process of the colloidal gold immunochromatographic test strip for paraquat is improved and optimized, so that the prepared test strip can rapidly and qualitatively detect paraquat in human urine within 15 minutes, and compared with the existing detection time of a professional detection mechanism which needs seven to eight hours and generally needs about one week for the detection report, the technical scheme has a qualitative leap in detection efficiency. And the immunochromatography test strip is used for detection, so that the cost is low, only about dozens of yuan is needed for detection once, and compared with thousands of yuan for instrument detection, the cost of detection is greatly reduced.
in addition, in the process of detecting the paraquat, because the metabolic pathway of the paraquat is metabolized through urine, the pH is adjusted by ammonia water before detection, so that the paraquat is free from the urine at a specific pH value and becomes a monomer to exist, and the detection of the paraquat is possible. The colloidal gold immunochromatographic assay rapid detection test strip prepared by the technical scheme has the advantages that the sensitivity can reach 50ng/mL, the lowest detection concentration can be compared favorably with large-scale instruments such as a high-phase liquid-phase mass spectrum combination instrument, the result is accurate and reliable, professional technicians and large-scale instruments are not needed, the detection operation is simple and convenient, the test strip is suitable for laboratories, is suitable for vast basic level personnel and large-scale detection, and has strong applicability.
further, in the step a, the preparation process of the colloidal gold solution comprises the steps of stirring and heating the aqueous solution of the gold chlorate to boiling, adding the aqueous solution of trisodium citrate, continuing stirring and heating until the solution is wine red, and cooling at room temperature to obtain the colloidal gold solution containing the colloidal gold particles with the particle size of 10-30 nm.
In the technical scheme, through research, the trisodium citrate aqueous solution is rapidly added into the gold chlorate aqueous solution once only when the colloidal gold solution is prepared, and rapid stirring is assisted, so that the particle size of colloidal gold particles can be effectively ensured, the particle size of the colloidal gold particles in the colloidal gold solution is uniform, the concentration of the prepared colloidal gold solution is further improved, the paraquat monoclonal antibody is not required to be concentrated in the later period when the paraquat monoclonal antibody is marked, and the preparation steps of the colloidal gold immunochromatographic test strip are simplified.
further, in the step b, the resuspension is a mixed solution of 0.02-0.1mol/LTris-HCl, bovine serum albumin with the mass concentration of 3% -8%, TritonX-100 with the mass concentration of 0.1% -0.3% and sucrose with the mass concentration of 5% -15% in equal volume ratio.
in the technical scheme, the composition of the resuspension can play a positive role in the sensitivity and the chromatography speed of the prepared test strip, so that paraquat in urine can smoothly pass through chromatography, and the sensitivity of the immunochromatographic test strip is further improved.
further, in the step b, the colloidal gold labeled paraquat monoclonal antibody solution is stored in a refrigerator at 4 ℃ for later use.
In the technical scheme, the marked paraquat monoclonal antibody solution is stored at low temperature, so that the stability of the colloidal gold solution marked by the paraquat monoclonal antibody can be maintained, and the failure is avoided.
Furthermore, when the colloidal gold solution is centrifuged in step b, the rotation speed is 9000-12000r/min, and the centrifugation time is 10-30 min.
the colloidal gold solution is centrifuged under the condition, so that the separation effect of the clear liquid and the precipitate in the colloidal gold solution can be ensured.
Further, in the step c, the gold spraying process is to spray colloidal gold solution on the glass fiber bonding pad according to the spraying amount of 6-15 mul/cm, and the glass fiber bonding pad after gold spraying is obtained after drying.
Through adjusting the volume of spouting gold, can guarantee the chromogenic effect and the sensitivity of colloidal gold immunochromatography test paper strip at the front end, in addition, the glass fiber bonding pad is easily obtained and stability can be high, can guarantee the stable combination of paraquat monoclonal antibody mark's colloidal gold solution, and choose for use glass fiber bonding pad, interact with heavy suspension, be favorable to the release of colloidal gold granule and the chromatography process of waiting to examine the sample.
Further, in the step d, the C thread packet is coated with goat anti-mouse IgG, and the concentration is 0.5-1.5 mg/ml; and (3) coating the T-shaped coil with the detection antigen of paraquat at the concentration of 0.5-2.0mg/ml, and drying to obtain the coated nitrocellulose membrane.
in the technical scheme, the sensitivity and the detection effect of the colloidal gold immunochromatographic test strip in the later period can be ensured by optimizing the coating substance and the substance concentration.
And step e, sticking the sample pad, the glass fiber combined pad after gold spraying, the coated nitrocellulose membrane and the absorbent paper on a polyvinyl chloride bottom plate in a sequentially connected mode, assembling, and cutting into strips to obtain the paraquat colloidal gold immunochromatography rapid detection test strip.
in the technical scheme, the assembled colloidal gold immunochromatographic test paper is cut into strips, so that the colloidal gold immunochromatographic test paper is convenient to store and use in later periods.
further, when reading the result, the test strip for quick detection of paraquat colloidal gold immunochromatography was vertically placed on the front of the observer with one end of the sample pad facing downward.
In the technical scheme, when a result is read, the test strip dropwise added with the detection sample is vertically placed, so that the dipping rate of the detection sample can be accelerated, the effective combination of the detection sample and the immunochromatography test strip can be ensured, and the detection precision is improved.
Detailed Description
The following is further detailed by way of specific embodiments:
Examples 1 to 8 are specific examples of the present invention, and parameters of the paraquat colloidal gold immunochromatographic rapid test strip prepared in each example are shown in table 1.
TABLE 1
the preparation method of the colloidal gold immunochromatographic test strip for paraquat in urine is illustrated by taking the example of example 1, and comprises the following steps:
Step a: preparing a colloidal gold solution: taking 80ml of chloroauric acid aqueous solution with the mass concentration of 0.008%, stirring and heating to boiling, quickly adding 3ml of trisodium citrate aqueous solution with the mass concentration of 0.8% at one time, continuously stirring and heating for 1h until the solution is wine red, and cooling at room temperature to obtain colloidal gold solution containing colloidal gold particles with the particle size of 18 nm;
step b: labeling paraquat monoclonal antibody with colloidal gold solution: centrifuging 8ml of the colloidal gold solution obtained in the step a at 10000r/min for 30min, discarding the supernatant, resuspending the precipitate by using 4ml of potassium carbonate solution with the pH value of 8.0, adding 0.04mg of paraquat monoclonal antibody into the colloidal gold solution, uniformly stirring, and standing for 2.0; dropwise adding 0.4ml of sterile bovine serum albumin with the mass concentration of 10%, stirring for 1.0 hour, and standing overnight at room temperature; centrifuging the marked colloidal gold solution at 10000r/min for 15min, removing supernatant, re-dissolving the precipitate with 4ml of resuspension (mixed solution of 0.05mol/LTris-HCl, bovine serum albumin with the mass concentration of 4%, TritonX-100 with the mass concentration of 0.1% and sucrose with the mass concentration of 5% in an equal volume ratio) to obtain a colloidal gold marked paraquat monoclonal antibody solution, and storing the colloidal gold marked paraquat monoclonal antibody solution in a refrigerator at 4 ℃ for later use;
Step c: spraying gold: b, spraying the colloidal gold-labeled paraquat monoclonal antibody solution obtained in the step b on a glass fiber bonding pad according to the spraying amount of 6 mu l/cm, and drying in a drying oven at 37 ℃ for 24 hours to obtain the glass fiber bonding pad after gold spraying;
Step d: coating a C line and a T line on the nitrocellulose membrane: coating a C line and a T line on a nitrocellulose membrane, wherein the C line is coated with goat anti-mouse IgG and the concentration is 1.0 mg/ml; the concentration of the T line wrapped by the detection antigen of paraquat is 2.0 mg/ml; drying in an oven at 37 ℃ for 24 hours to obtain the coated nitrocellulose membrane, wherein a detection line T line is close to one end of the sample pad;
step e: assembling the colloidal gold immune test strip: and (3) pasting the sample pad, the glass fiber combined pad after gold spraying, the coated nitrocellulose membrane and the absorbent paper on a polyvinyl chloride bottom plate in a sequentially connected mode for assembly, and cutting into strips to obtain the paraquat colloidal gold immunochromatography rapid detection test strip.
The test strips prepared in examples 1 to 8 each include a base plate, the base plate is sequentially connected with a sample pad, a conjugate pad after gold spraying, a coated nitrocellulose membrane and absorbent paper, the conjugate pad after gold spraying is sprayed with a gold-labeled paraquat monoclonal antibody solution with a gold particle size of 18nm, and the gold spraying amount is 6. mu.l/cm, 10. mu.l/cm, 15. mu.l/cm, 10. mu.l/cm, 6. mu.l/cm, 10. mu.l/cm, 15. mu.l/cm and 10. mu.l/cm, respectively.
Preparing biological sample urine with different paraquat concentrations, wherein the concentration of paraquat in the biological sample urine is 1000ng/ml, 800ng/ml, 600ng/ml, 400ng/ml, 200ng/ml, 100ng/ml, 90ng/ml, 80ng/ml, 70ng/ml, 60ng/ml, 50ng/ml, 40ng/ml and 30ng/ml respectively. The colloidal gold immunochromatographic rapid detection test strip prepared in the examples 1 to 8 is used for detection, and meanwhile, paraquat detected by liquid-mass combination (comparative example 1) in the prior art is used as a comparative example for detection, and each concentration is subjected to three repeated tests.
The detection steps of the paraquat colloidal gold immunochromatographic assay test strip in urine are as follows:
(1) Taking 1ml of urine of a biological sample, adding about 100 mu l of ammonia water solution with the mass concentration of 25%, measuring the pH value to be 7.5 by using a pH test strip, and standing for 5 minutes;
(2) Sucking 100 mu l of the sample solution to be detected obtained in the step (1) by using a dropper, dripping the sample solution on a sample pad of a test strip, and timing after dripping the sample;
(3) After dropping the sample for 15min, reading the result, and during reading, vertically placing the test strip on the front side of an observer in a manner that one end of the sample pad faces downwards;
(4) and (5) judging a result: the C line of the quality control line is displayed as a red line, when the T line is developed, the result is negative, and the concentration of paraquat in the sample to be detected is less than 50 ng/ml; when the T line does not develop color, the result is positive, and the concentration of paraquat in the sample to be detected is more than 50 ng/ml; the control line C line is not colored, and the test paper strip is invalid.
the results of urine testing of biological specimens using the examples and comparative examples are shown in Table 2:
TABLE 2
Note: + positive result, detecting; negative results, no detection.
Table 3 units of assay time comparison: min (detection time is the time when the test strip begins to display the result)
The colloidal gold immunochromatographic test strip prepared by the technical scheme can accurately detect paraquat in urine by combining tables 2 and 3, and the result shows that the detection sensitivity is 50 mu g/L, the lowest detection concentration can be completely comparable with that of the traditional large-scale instrument detection equipment, the technical bias that the detection of paraquat in the prior art can be completed only by large-scale equipment is overcome, the prepared colloidal gold immunochromatographic test strip is accurate and reliable in detection result, professional technicians and large-scale instrument equipment are not needed, the detection operation is simple and convenient, the colloidal gold immunochromatographic test strip is suitable for laboratories, is suitable for vast basic level personnel and large-scale detection, and is high in applicability and low in detection cost. In the aspect of detection time, the detection of results can be completed within 15min by adopting the colloidal gold immunochromatographic test strip, the detection time is shortened by more than 30 times compared with the traditional liquid chromatography-mass spectrometry detection method, and the method has important significance in clinical rapid screening of paraquat. In addition, the paraquat colloidal gold immunochromatographic test strip can also be used for on-site rapid detection of emergency treatment of sudden poisoning and family personal screening detection, and has the advantages of wide application range and low operation difficulty.
The foregoing is merely an example of the present invention and common general knowledge in the art of designing and/or characterizing particular aspects and/or features is not described in any greater detail herein. It should be noted that, for those skilled in the art, without departing from the technical solution of the present invention, several variations and modifications can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicability of the patent. The scope of the claims of the present application shall be determined by the contents of the claims, and the description of the embodiments and the like in the specification shall be used to explain the contents of the claims.
Claims (10)
1. the preparation method of the colloidal gold immunochromatographic test strip for paraquat in urine is characterized by comprising the following steps of:
Step a: preparing a colloidal gold solution: preparing a colloidal gold solution containing colloidal gold particles with the particle size of 10-30nm by taking a chloroauric acid aqueous solution and a trisodium citrate aqueous solution as raw materials;
Step b: labeling paraquat monoclonal antibody with colloidal gold solution: centrifuging the colloidal gold solution in the step a, then removing supernatant, and resuspending the precipitate by using potassium carbonate solution with the pH of 8.0-8.5; adding 0.04-0.1mg of paraquat monoclonal antibody into the colloidal gold solution, stirring uniformly and standing; dropwise adding sterile bovine serum albumin with the mass concentration of 8-12%, stirring, standing overnight at room temperature to obtain a labeled colloidal gold solution; centrifuging the labeled colloidal gold solution again, discarding the supernatant, and re-dissolving the precipitate with 4-6mL of heavy suspension to obtain a colloidal gold-labeled paraquat monoclonal antibody solution;
Step c: spraying gold;
Step d: coating the nitrocellulose membrane with a C line and a T line;
Step e: and (5) assembling the colloidal gold immune test strip.
2. The method for preparing the colloidal gold immunochromatographic test strip for paraquat in urine according to claim 1, which is characterized in that: in the step a, the preparation process of the colloidal gold solution comprises the steps of stirring and heating a gold chlorate aqueous solution to boiling, adding a trisodium citrate aqueous solution, continuing stirring and heating until the solution is wine red, and cooling at room temperature to obtain the colloidal gold solution containing the colloidal gold particles with the particle size of 10-30 nm.
3. The method for preparing the colloidal gold immunochromatographic test strip for paraquat in urine according to claim 2, which is characterized in that: in the step b, the resuspension is a mixed solution of 0.02-0.1mol/LTris-HCl, bovine serum albumin with the mass concentration of 3% -8%, TritonX-100 with the mass concentration of 0.1% -0.3% and sucrose with the mass concentration of 5% -15% in an equal volume ratio.
4. the method for preparing the colloidal gold immunochromatographic test strip for paraquat in urine according to claim 3, which is characterized in that: and in the step b, storing the colloidal gold-labeled paraquat monoclonal antibody solution in a refrigerator at 4 ℃ for later use.
5. the method for preparing the colloidal gold immunochromatographic test strip for paraquat in urine according to claim 4, which is characterized in that: when the colloidal gold solution is centrifuged in the step b, the revolution is 9000-12000r/min, and the centrifugation time is 10-30 min.
6. The method for preparing the colloidal gold immunochromatographic test strip for paraquat in urine according to claim 5, which is characterized in that: and c, spraying the colloidal gold solution on the glass fiber bonding pad according to the spraying amount of 6-15 mu l/cm in the gold spraying process, and drying to obtain the glass fiber bonding pad after gold spraying.
7. the method for preparing the colloidal gold immunochromatographic test strip for paraquat in urine according to claim 6, which is characterized in that: in the step d, coating goat anti-mouse IgG on the C thread, wherein the concentration is 0.5-1.5 mg/ml; and (3) coating the T-shaped coil with the detection antigen of paraquat at the concentration of 0.5-2.0mg/ml, and drying to obtain the coated nitrocellulose membrane.
8. the method for preparing the colloidal gold immunochromatographic test strip for paraquat in urine according to claim 7, which is characterized in that: and e, pasting the sample pad, the glass fiber combined pad after gold spraying, the coated nitrocellulose membrane and the absorbent paper on a polyvinyl chloride bottom plate in a sequentially connected mode for assembly, and cutting into strips to obtain the paraquat colloidal gold immunochromatography rapid detection test strip.
9. the method for using the colloidal gold immunochromatographic strip for paraquat in urine as claimed in any one of claims 1 to 8, which comprises the following steps:
Step (1): taking 1-3ml of biological specimen urine, adding ammonia water, adjusting the pH value to 7.0-8.0, and standing to obtain a sample solution to be detected;
Step (2): dropwise adding the sample solution to be detected on a paraquat colloidal gold immunochromatographic rapid detection test strip, and timing;
and (3): reading the result after timing;
and (4): and (5) judging a result: the C line of the quality control line is displayed as a red line, when the T line is developed, the result is negative, and the concentration of paraquat in the sample to be detected is less than 50 ng/ml; when the T line does not develop color, the result is positive, the concentration of paraquat in the sample to be detected is more than 50ng/ml, the quality control line C line does not develop color, and the test strip judges that the test strip is invalid.
10. The use method of the colloidal gold immunochromatographic strip for paraquat in urine according to claim 9, is characterized in that: when reading the result, the paraquat colloidal gold immunochromatographic rapid detection test strip is vertically placed on the front of an observer in a manner that one end of the sample pad faces downwards.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910942583.2A CN110554193A (en) | 2019-09-30 | 2019-09-30 | Preparation method of colloidal gold immunochromatographic test strip for paraquat in urine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910942583.2A CN110554193A (en) | 2019-09-30 | 2019-09-30 | Preparation method of colloidal gold immunochromatographic test strip for paraquat in urine |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110554193A true CN110554193A (en) | 2019-12-10 |
Family
ID=68741993
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910942583.2A Pending CN110554193A (en) | 2019-09-30 | 2019-09-30 | Preparation method of colloidal gold immunochromatographic test strip for paraquat in urine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110554193A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110940805A (en) * | 2020-02-14 | 2020-03-31 | 北京纳百生物科技有限公司 | Immune colloidal gold homogeneous phase labeling method |
| CN112986566A (en) * | 2021-03-29 | 2021-06-18 | 浙江理工大学 | Colloidal gold labeling method for silkworm monoclonal antibody for identifying silk fabrics |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101324575A (en) * | 2008-07-10 | 2008-12-17 | 江南大学 | Preparation and application of paraquat semi-quantitative rapid test reagent strip |
| CN101493458A (en) * | 2009-03-03 | 2009-07-29 | 周坚 | Paraquat-glyphosate dual detecting card and processing method of test sample thereof |
| CN106568904A (en) * | 2015-10-13 | 2017-04-19 | 江苏维赛科技生物发展有限公司 | Immunity colloid gold detection card of paraquat and preparation method thereof |
-
2019
- 2019-09-30 CN CN201910942583.2A patent/CN110554193A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101324575A (en) * | 2008-07-10 | 2008-12-17 | 江南大学 | Preparation and application of paraquat semi-quantitative rapid test reagent strip |
| CN101493458A (en) * | 2009-03-03 | 2009-07-29 | 周坚 | Paraquat-glyphosate dual detecting card and processing method of test sample thereof |
| CN106568904A (en) * | 2015-10-13 | 2017-04-19 | 江苏维赛科技生物发展有限公司 | Immunity colloid gold detection card of paraquat and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 孙秀兰等: "粮食中百草枯残留的金标免疫层析检测方法研究", 《分析测试学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110940805A (en) * | 2020-02-14 | 2020-03-31 | 北京纳百生物科技有限公司 | Immune colloidal gold homogeneous phase labeling method |
| CN110940805B (en) * | 2020-02-14 | 2022-11-29 | 北京纳百生物科技有限公司 | Immune colloidal gold homogeneous phase labeling method |
| CN112986566A (en) * | 2021-03-29 | 2021-06-18 | 浙江理工大学 | Colloidal gold labeling method for silkworm monoclonal antibody for identifying silk fabrics |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111398588A (en) | Use method of immunochromatography kit for rapidly detecting novel coronavirus N protein | |
| CN111398583A (en) | Kit for detecting novel coronavirus N protein and application thereof | |
| CN102177437B (en) | Reference control for cell by cell analysis | |
| US20130084580A1 (en) | Chromatographic kit and chromatography method | |
| JP2000514196A (en) | Determination of glycohemoglobin (%) | |
| CN111751527A (en) | Novel coronavirus IgG and IgM antibody detection kit and preparation method and application thereof | |
| JPH08501145A (en) | Solid-phase immunoassay | |
| CN111308072A (en) | Colloidal gold immunochromatography kit for rapidly detecting novel coronavirus IgG antibody and preparation method thereof | |
| CN113325171B (en) | Kit for detecting human body erythrocyte folic acid content, detection method and application | |
| CN110554193A (en) | Preparation method of colloidal gold immunochromatographic test strip for paraquat in urine | |
| CN113834944B (en) | Quantum dot fluorescence detection method for folic acid in red blood cells | |
| CN104865247A (en) | Application of color development method based on nanogold aggregation to immunodetection | |
| KR101451733B1 (en) | Labeling agent for aflatoxin B1 detection and the kit for detecting aflatoxin B1 comprising thereof | |
| CN110441537B (en) | Method for measuring content of 25-hydroxy vitamin D | |
| US11141732B2 (en) | Kit for quickly detecting lead content in sample | |
| US4714672A (en) | Immunoassays involving complement lysing of chromophore containing microcapsules | |
| CN116990510A (en) | Adenovirus antigen reagent strip for detecting ocular secretion based on colloidal gold method and preparation method thereof | |
| US20210096075A1 (en) | Kit for quickly detecting cadmium content in sample | |
| CN110596378A (en) | Multichannel universal chromatography method for detecting small molecules, test strip and kit | |
| CN114675028A (en) | Preparation method and application of fenitrothion nano antibody-colloidal gold marker | |
| Sidki et al. | Direct determination of phenobarbital in serum or plasma by polarization fluoroimmunoassay | |
| CN104459152A (en) | Rapid sperm concentration detection method based on colloidal gold immunochromatograohic assay technology | |
| EP1256802A1 (en) | Method for examination of feces occult blood | |
| CN112180080B (en) | Small molecule triple immunochromatography detection method, test strip and kit | |
| JP4273111B2 (en) | Detection apparatus and detection method |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20191210 |
|
| RJ01 | Rejection of invention patent application after publication |