CN112114139A - Novel coronavirus IgM-IgA-IgG antibody colloidal gold detection kit - Google Patents
Novel coronavirus IgM-IgA-IgG antibody colloidal gold detection kit Download PDFInfo
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Abstract
The invention provides a novel coronavirus IgM-IgA-IgG antibody colloidal gold detection kit, which is characterized in that card strips for detecting IgM antibody, IgA antibody and IgG antibody are respectively and parallelly arranged in a card shell to be assembled into a detection card; the gold-labeled areas of the coated plates of the three card strips are respectively coated with colloidal gold-labeled anti-human IgM, colloidal gold-labeled anti-human IgA and colloidal gold-labeled anti-human IgG, the detection areas are coated with novel coronavirus recombinant antigens, and the quality control areas are coated with goat anti-mouse IgG; or respectively coating anti-human IgM antibody, anti-human IgA antibody and anti-human IgG antibody in different areas of the nitrocellulose membrane as detection areas, coating streptavidin at positions above three detection lines as quality control lines, and fixing the colloidal gold-labeled recombinant antigen and biotinylated bovine serum albumin in the gold-labeled area. Wherein the novel coronavirus recombinant antigen is S-RBD protein. The invention realizes the joint detection of the novel coronavirus IgM-IgA-IgG antibody and further improves the detection sensitivity.
Description
Technical Field
The invention belongs to the technical field of immunoassay detection, and particularly relates to a novel coronavirus IgM-IgA-IgG antibody colloidal gold detection kit.
Background
The coronavirus belongs to single-stranded positive-strand RNA virus, and there are 6 kinds of coronavirus known to infect humans, namely HCoV-229E, HCoV-OC43, SARSr-CoV, HCoV-NL63, HCoV-HKU1 and MERSR-CoV. The novel coronavirus (2019-nCoV) belongs to the 7 th species.
The novel coronavirus pneumonia is an acute infectious pneumonia, and the pathogen of the novel coronavirus is a novel coronavirus which is not found in human before, namely 2019 novel coronavirus. Transmission via respiratory droplets and intimate contact is the primary transmission route, and there is the potential for transmission via aerosols during prolonged exposure to high concentrations of aerosols in a relatively closed environment. The initial symptoms of the patients are mostly fever, hypodynamia and dry cough, and the patients gradually show severe manifestations such as dyspnea and the like. The prognosis is good in most patients and acute respiratory distress syndrome or septic shock may occur in some severe cases and even death.
The novel coronavirus SARS-COV-2 is known as a mucosa-targeted virus, and IgM, IgA and IgG antibodies appear at different infection stages. However, the novel coronavirus IgM-IgA-IgG antibody joint detection is lacked at present, and a method for quickly detecting the novel coronavirus IgM-IgA-IgG antibody joint detection is provided, so that the detection sensitivity is further improved.
Disclosure of Invention
In view of this, the invention aims to provide a novel coronavirus IgM-IgA-IgG antibody colloidal gold detection kit to realize the joint detection of the novel coronavirus IgM-IgA-IgG antibody and further improve the detection sensitivity.
A novel coronavirus IgM-IgA-IgG antibody colloidal gold detection kit is characterized in that card strips for detecting IgM antibodies, IgA antibodies and IgG antibodies are respectively arranged in parallel in a card shell to be assembled into a detection card; the gold-labeled areas of the coated plates of the three card strips are respectively coated with colloidal gold-labeled anti-human IgM, colloidal gold-labeled anti-human IgA and colloidal gold-labeled anti-human IgG, the detection areas are coated with novel coronavirus recombinant antigens, and the quality control areas are coated with goat anti-mouse IgG; wherein the novel coronavirus recombinant antigen is S-RBD protein.
Preferably, the preparation method of the colloidal gold-labeled anti-human IgA, colloidal gold-labeled anti-human IgM or colloidal gold-labeled anti-human IgG comprises the following steps:
adding K to homogeneous colloidal gold solution2CO3Adjusting the pH to 6.0-10.0, adjusting the pH to 6.5-8.0 when coupling an antibody, adjusting the pH to 8.0-10.0 when coupling an antigen, adding anti-human IgA to the solution with the adjusted pH to ensure that the protein content of the anti-human IgA is 15-60 mu g/mL, coupling for 0.5-4 hours, adding a stop solution to the solution to stop the coupling reaction, wherein the final concentration of the stop solution is 1-1.5%; centrifuging after 0.5-2 hours after the reaction is ended, centrifuging for 15-30min at the centrifugal parameter of 12000-18000RPM, discarding the supernatant, redissolving the precipitate by using a redissolution, and storing at 2-8 ℃ for later use; the IgM and IgG antibodies are labeled by the same method as IgA.
The invention also provides a novel coronavirus IgM-IgA-IgG antibody colloidal gold detection kit, which comprises a detection card provided with a coating plate and a sample pad, wherein colloidal gold particles in a gold mark area of the coating plate coat novel coronavirus recombinant antigens and biotinylated bovine serum albumin, different positions of a detection area are respectively coated with anti-human IgM, anti-human IgA and anti-human IgG, and a quality control area is coated with streptavidin; wherein the novel coronavirus recombinant antigen is S-RBD protein.
Preferably, the preparation method of the novel coronavirus recombinant antigen coated by the colloidal gold particles comprises the step of coating a colloidal gold solution with 0.1M K2CO3Adjusting pH to 8.5-9.0, mixing thoroughly for 5-10 min, adding novel coronavirus recombinant antigen to the mixture to a final concentration of 10-30 μ g/mL, adding bovine serum albumin to the mixture to a final concentration of 0.8-1.2% after 10-30 min, discarding the supernatant after 30-60 min, 10000RPM, centrifuging for 10-15min, discarding the supernatant, and redissolving the precipitate with a redissolution; spraying the colloidal gold conjugate on a glass fiber membrane according to the spraying amount of 45-60 muL/cm, and drying at 35-45 ℃.
Preferably, the sample pad is coated with a sample treatment solution, and the sample treatment solution is tris buffer solution which contains BSA with the mass concentration of 0.5-1.5%, casein with the mass concentration of 0.5-1.5%, sucrose with the mass concentration of 0.3-1.2%, trehalose with the mass concentration of 0.1-1.2%, Procline with the volume concentration of 0.05-0.3%, and tween-20 with the volume concentration of 0.05-0.5% and has the pH value of 7.0-7.6; the preparation method of the sample pad comprises spraying or soaking the sample pad treating solution onto or into the glass fiber film or polyester film at 45-85 microliter per square centimeter, and oven drying at 37-45 deg.C for 8-16 hr.
Preferably, the colloidal gold solution is prepared by preparing chloroauric acid into a solution with a mass-to-volume ratio of 1% -10%, adding the solution with a final concentration of one ten thousandth to four ten thousandth into boiled purified water, continuing to boil for 2-5 minutes, adding 0.1-0.2M sodium citrate into 100mL chloroauric acid solution in an amount of 500-800 microliters, and continuing to stir and heat for 10-20 minutes to prepare the colloidal gold solution with uniform particle size.
Preferably, the kit further comprises a sample diluent and a 0.05M PB buffer solution, wherein the pH value is 8.0, and 0.5-3% by mass and volume of NaCl and 0.5-5% by mass and volume of sucrose are added into the sample diluent.
The invention also provides application of the kit in preparation of a novel coronavirus IgM-IgA-IgG antibody detection reagent.
The conjugate pad is a module to which the labeled conjugate is attached, and the treatment solution for treating the conjugate pad is pH 8.0-8.5, 0.01-0.05M tris buffer, 0.1-0.3% tween-20.
The coating buffer solution for diluting the antigen-antibody raw material for coating is 0.01-0.05M buffer solution with pH of 7.4-8.5, 0.1-3% sucrose, 0.1-1% casein.
The principle of the invention is that,
the detection card is assembled by respectively putting card strips for detecting IgM antibody, IgA antibody and IgG antibody into a card shell in parallel, the kit adopts an immunochromatography method, a blood sample to be detected is added on the detection card, novel coronavirus IgM/IgA/IgG antibody and non-novel coronavirus IgM/IgA/IgG antibody in the sample are combined with anti-human IgM/IgA/IgG marked by colloidal gold to form a novel coronavirus IgM/IgA/IgG antibody-anti-human IgM/IgA/IgG antibody-colloidal gold compound, the non-novel coronavirus IgM/IgA/IgG antibody-anti-human IgM/IgA/IgG antibody-colloidal gold compound, the immune compound moves forwards along the detection card under the capillary action, and the novel coronavirus IgM/IgA/IgG antibody-anti-human IgM/IgA/IgG antibody-colloidal gold compound can be coated on a new detection area on a membrane And (3) capturing the recombinant antigen of the coronavirus to form a purple red strip, continuously moving the non-novel coronavirus IgM/IgA/IgG antibody-antihuman IgM/IgA/IgG-colloidal gold antibody compound forwards, combining with a goat anti-mouse in a quality control area to form a purple red strip, and respectively prompting that the novel coronavirus IgM/IgA/IgG antibody is positive corresponding to the condition that three window strips appear.
When a sample contains novel coronavirus specific antibodies IgM and (or) IgA and (or) IgG, the detection card is combined with gold-labeled recombinant protein to form a specific antibody IgM and (or) IgA and (or) IgG-novel coronavirus recombinant protein-colloidal gold compound, the compound continuously moves forwards under the capillary action and moves to the position coated with an anti-human IgM strip, and the specific antibody IgM-novel coronavirus recombinant protein-colloidal gold compound is captured to form a purplish red strip to prompt that the antibody is positive; on the contrary, when the sample does not contain the novel coronavirus antibody IgM, a color development band cannot be formed on the anti-human IgM band, and the antibody is indicated to be negative. The detection principle of the novel coronavirus specific antibody IgA/IgG on the detection card is the same as that of the IgM described above.
Compared with the prior art, the kit has the following advantages: the three-item joint inspection can further improve the detection sensitivity compared with single-item detection and two-item joint inspection.
Drawings
FIG. 1 is a test strip condition analysis in accordance with one embodiment;
FIG. 2 is a test strip analysis of example two.
Detailed Description
Unless defined otherwise, technical terms used in the following examples have the same meanings as commonly understood by one of ordinary skill in the art to which the present invention belongs. The test reagents used in the following examples, unless otherwise specified, are all conventional biochemical reagents; the experimental methods are conventional methods unless otherwise specified.
The present invention will be described in detail with reference to examples.
The joint detection of the three antibodies can further improve the detection sensitivity and can be realized by the following method, the detection card can be assembled and formed by respectively preparing IgM/IgA/IgG card strips and one card shell which can be simultaneously filled with the three card strips in parallel. Under this form, this kit is including being equipped with the detection card of peridium board and sample pad, and the gold mark district of peridium board is wrapped up respectively and is had anti human IgM/IgA/IgG of colloidal gold mark, and the detection area is wrapped up and is had novel coronavirus recombination antigen, and quality control district is wrapped up goat anti mouse IgG, need carry out the application of sample operation to every detection card during the use, and the testing result independently demonstrates in every detection card window. Or anti-human IgM/IgA/IgG can be respectively coated on the same card strip in the detection area, the quality control area is coated with Streptavidin (SA), and the colloidal gold particles in the gold label area are coated with the novel coronavirus recombinant antigen. When in use, only one sample is needed to be added, and the result is judged according to the marked IgM/IgA/IgG position on the card surface.
In both of the above two preparation forms, a reaction region and a quality control region of the detection card, a gold label region, and a sample pad are indispensable.
The sample pad is coated with a sample treatment solution, and the sample treatment solution is a Tris buffer solution which contains BSA with the mass concentration of 1%, casein with the mass concentration of 0.5%, sucrose with the mass concentration of 0.5%, trehalose with the mass concentration of 1.0%, Proclin 300 with the volume concentration of 0.05%, tween-20 with the volume concentration of 0.08% and has the pH value of 7.4; the preparation method of the sample pad comprises spraying or soaking the sample pad treatment solution with 60 microliter per square centimeter on glass fiber film or polyester film, and oven drying at 37 deg.C for 12 hr.
Example one
A novel coronavirus IgM-IgA-IgG antibody colloidal gold detection kit is characterized in that card strips for detecting IgM antibodies, IgA antibodies and IgG antibodies are respectively arranged in parallel in a card shell to be assembled into a detection card; the gold-labeled areas of the coated plates of the three card strips are respectively coated with colloidal gold-labeled anti-human IgM, colloidal gold-labeled anti-human IgA and colloidal gold-labeled anti-human IgG, the detection areas are coated with novel coronavirus recombinant antigens, and the quality control areas are coated with goat anti-mouse IgG; wherein the novel coronavirus recombinant antigen is S-RBD.
The preparation of colloidal gold-labeled anti-human IgA comprises the step of adding K to a homogeneous colloidal gold solution2CO3Adjusting the pH value to 8.0, adding anti-human IgA to the solution with the adjusted pH value to enable the protein content to be 25 mu g/mL, coupling for 0.5 hour, adding a stop solution to the solution, and stopping the coupling reaction; centrifuging after 0.5 hour after reaction termination, wherein the centrifugation parameter is 12000RPM, centrifuging for 20min, discarding the supernatant, redissolving the precipitate with redissolving solution, and storing at 2-8 ℃ for later use. The colloidal gold labeled IgM/IgG method is the same as IgA.
The stop solution is bovine serum albumin or skim milk powder with the mass concentration of 10%; the compound solution is 0.02M PB buffer solution, the pH value is 7.4, and the buffer solution is added with 0.9 mass volume percent of NaCl, 0.8 mass volume percent of trehalose and 1.5 mass volume percent of PEG 20000; bovine serum albumin mass volume ratio of 1.5%, Triton X-100 volume ratio of 0.5%.
The colloidal gold solution is prepared by preparing chloroauric acid into a solution with the mass volume ratio of 10%, adding the solution with the final concentration of chloroauric acid being two-fifths of the mass volume ratio into boiled purified water, continuously boiling for 3 minutes, adding 580 microliters of sodium citrate into each 100mL of chloroauric acid solution, continuously stirring and heating for 10 minutes, cooling to room temperature, thus preparing the colloidal gold solution with uniform particle size, and storing at 2-8 ℃ for later use.
Preparing the detection card, namely paving the colloidal gold-labeled anti-human IgA on a labeling pad uniformly, and drying for 12 hours at 37 ℃ to obtain the labeling pad; preparing the novel coronavirus recombinant antigen and goat anti-mouse IgG into solutions with certain concentrations respectively, and coating the solutions at the positions of a T line and a C line respectively by using a film-cutting gold spraying instrument at 37 ℃ for drying to obtain coated nitrocellulose membranes; and sequentially overlapping and sticking the absorbent paper, the nitrocellulose membrane, the marking pad, the blood filtering membrane and the sample pad on the PVC plate.
The blood filter membrane is used after being treated by an anti-RBC antibody, the treatment concentration is 0.1mg/mL, the treatment capacity is 60 microliters per square centimeter, and the blood filter membrane is dried for 12 hours at the temperature of 37 ℃. Can be used for intercepting red blood cells in blood sample.
Also comprises sample diluent, 0.05M PB buffer solution with pH of 8.0, and NaCl with mass-volume ratio of 0.5-3% and sucrose with mass-volume ratio of 0.5-5%.
The kit comprises the following reaction steps:
procedure of experiment
1 if the sample is stored in a refrigerated or frozen state, the sample to be tested and the required reagents are removed from the storage conditions and allowed to equilibrate to room temperature (20-30 ℃). Fully and uniformly mixing the sample to be detected after melting;
2 when in preparation for detection, opening the aluminum foil bag from the tear, taking out the detection card, and horizontally placing the detection card on a horizontal desktop;
marking a sample number on the detection card;
and 4, sucking 10 mu L of the sample to be detected from the sample tube by using a pipette or a dropper, and dripping 100 mu L of buffer solution to ensure that no bubbles exist in the operation process.
5, counting time for 15 minutes, judging the result, and after 20 minutes, the judging result is invalid.
Interpretation of test results
1, when only a quality control line C appears, the corresponding (IgM/IgA/IgG) detection zone detection line T is not colored, which indicates that no novel coronavirus antibody is detected, and the result is negative, as shown in (a) in FIG. 1 (it should be noted that the schematic diagram does not include all detection conditions, and the result corresponding to the detection zone is understood to correspond to the negative and positive of the respective immune type antibody);
2, a positive result, when the quality control line C and the detection line T of the corresponding (IgM/IgA/IgG) detection area both appear, indicating that the antibodies (IgM and (or) IgA and (or) IgG) of the corresponding type of the novel coronavirus antibody are detected, as shown in fig. 1 (b) (it should be noted that the schematic diagram does not include all detection conditions, and it should be understood that the result of the corresponding detection area corresponds to the negative and positive of the respective immune antibody);
(iii) a null result, wherein if the quality control line C is not observed, the null result is obtained regardless of whether T is present or not, and the re-detection is performed as shown in (C/d) of FIG. 1 (it should be noted that the schematic does not include all detection cases, and the result corresponding to the detection region should be understood as negative or positive with respect to the respective immunotype antibody)
Through nucleic acid detection, 200 cases are diagnosed, 300 cases are excluded, the detection is carried out by using a novel coronavirus antibody IgM/IgA/IgG colloidal gold kit (card-discharging mode), and the detection result takes the positive nucleic acid detection as the reference. The detection sensitivity and the analysis specificity are evaluated, and test results show that the detection sensitivity of the single IgM is 83.64%, the detection sensitivity of the single IgG positive product is 85.1%, the detection sensitivity of the single IgA is 90.91, the detection sensitivity of the IgM/IgG joint detection is 92.73%, the detection sensitivity is higher than that of a single antibody type, and when the IgM/IgA/IgG joint detection is carried out, the detection sensitivity can reach 96.36%, so that the detection sensitivity is further improved. . The three-strip combined detection can further improve the positive detection rate compared with single antibody detection or two-antibody combined detection. The specific detection results are all over 95 percent, and the performance is good.
Summary of IgM/IgA/IgG Combined test (Row card mode)
Example two
This embodiment is for on the different regions of wrapping the detection zone to same card strip, can save the operation of assembling three detection membrane strips, only need install a membrane strip to plastics card shell can.
Compared with the above (card arrangement) embodiment, the most important difference is the coating and labeling process, in this embodiment, the detection area is coated with anti-human IgM/anti-human IgA/anti-human IgG, and the quality control area is coated with streptavidin.
The 0.01M PB buffer solution with the pH value of 8.0 of the coating buffer solution contains 1% of sucrose by mass and 0.1% of casein by mass.
And (3) diluting the anti-human IgM/anti-human IgA/anti-human IgG by using a coating buffer solution, respectively diluting the antibody content to 1.0mg/mL, and diluting the streptavidin content to 1.0mg/mL by using the coating buffer solution.
The reaction area and the quality control area of NC membrane (IgM/IgA/IgG) were streaked at 0.75uL/cm, and the coated plate was oven-dried at 37 ℃ for 16 hours.
In the binding pad area of the test card, there were colloidal gold conjugates of the novel coronavirus recombinant protein, and colloidal gold conjugates of biotinylated bovine serum albumin.
The novel colloidal gold conjugate of coronavirus recombinant protein is prepared by the following process:
mixing the colloidal gold solution with 0.1M K2CO3Adjusting pH to 9.0, mixing thoroughly for 5min, adding new coronavirus recombinant antigen to the mixture to a final concentration of 10 μ g/mL, adding bovine serum albumin to the mixture at a final concentration of 1% after 30min, discarding the supernatant after 30min, performing centrifugation at 10000RPM for 15min, discarding the supernatant, and redissolving the precipitate with a redissolution. The colloidal gold conjugate was sprayed on a glass fiber membrane in a spraying amount of 60. mu.L/cm, and dried at 37 ℃.
And (3) overlapping the dried combination pad and the sample pad in sequence and adhering the combination pad and the sample pad to a coated plate coated with anti-human IgM/IgA/IgG.
Cutting into 3-4mm strips, and placing into plastic card shell. NaCl solution with the mass fraction of 0.9% is prepared, and triton X-100 with the volume ratio of 0.1% is added as sample diluent.
And (3) testing: 10 μ L of the sample was added dropwise to the assay well followed by 80-100 μ L of sample diluent. And (3) carrying out forward chromatography along with the sample on the colloidal gold conjugate, wherein when the sample contains a corresponding specific antibody, the specific antibody can be combined with a recombinant antigen on the colloidal gold conjugate, and in the forward chromatography process, if the sample contains the specific IgM antibody, the specific IgM antibody is combined with coated anti-human IgM in an IgM reaction area to form a positive reaction condition, otherwise, no reaction condition is generated in the corresponding reaction area.
The results are explained as follows:
1, when only a quality control line C appears, the corresponding (IgM/IgA/IgG) detection zone detection line T is not colored, which indicates that no novel coronavirus antibody is detected, and the result is negative, as shown in (a) in FIG. 2 (it should be noted that the schematic diagram does not include all detection conditions, and the result corresponding to the detection zone is understood to correspond to the negative and positive of the respective immune type antibody);
2, a positive result, when the quality control line C and the detection line T of the corresponding (IgM/IgA/IgG) detection area both appear, indicating that the antibody (IgM and/or IgA and/or IgG) of the corresponding type of the novel coronavirus antibody is detected, as shown in (b) in fig. 2 (it should be noted that the schematic diagram does not include all detection cases, and it should be understood that the result of the corresponding detection area corresponds to the negative and positive of the respective immune antibody);
(iii) a null result, wherein if the quality control line C is not observed, the null result is obtained regardless of whether T is present or not, and the re-detection is performed as shown in FIG. 2 (C/d) (it should be noted that the schematic does not include all detection cases, and the result corresponding to the detection region should be understood as negative or positive with respect to the respective immunotype antibody)
Verification by equivalence of multi-strip detection mode and card arrangement mode
And simultaneously detecting a group of clinical samples by using a multi-strip joint detection mode detection card and a row card joint detection mode detection card, and confirming the detection consistency of the two forms by chi-square detection and kappa consistency analysis of the detection result.
The specific antibody IgM is subjected to statistical analysis by a card arrangement form and a multi-strip form detection result, a chi-square test result P is 1.000 and is more than 0.5, and the two methods have no significant difference; kappa is 0.976 ≧ 0.75, and the consistency of the kappa and the beta-cyclodextrin is good;
the statistical analysis is carried out on the detection results of the specific antibody IgA in a card arraying mode and a multi-strip mode, the chi-square test result P is 1.000 to 0.5, and the two methods have no significant difference; the kappa is 1.0 ≧ 0.75, and the diagnosis is consistent;
the specific antibody IgM is subjected to statistical analysis by a card arrangement form and a multi-strip form detection result, a chi-square test result P is 1.000 and is more than 0.5, and the two methods have no significant difference; kappa is 0.955 ≧ 0.75, and the consistency of the kappa and the beta-cyclodextrin is good;
2019-nCOV IgM row card form and multi-strip form detection card detection results
| 2019-nCOV IgM | Row card positive test | Row card detection negative | Total up to |
| Multiple line Pattern test Positive | 27 | 1 | 28 |
| Multiple line pattern detection negative | 0 | 82 | 82 |
| Total up to | 27 | 83 | 110 |
2019-nCOV IgM row card form and multi-strip form detection card detection results
Consistency analysis of the detection results of 2019-nCOV IgM row card format and the detection card of the multiple strip format kappa
2019-nCOV IgA row card type and multi-strip type detection card detection results
Chi-square test of 2019-nCOV IgA row card form and multi-strip form detection card detection results
Consistency analysis of the detection results of 2019-nCOV IgA arrayed cards and multi-strip-shaped detection cards
2019-nCOV IgG test card form and multi-strip form test card test result four-grid table
Chi-square test of detection results of 2019-nCOV IgG row card format and multi-strip format detection cards
Detection result kappa consistency analysis of 2019-nCOV IgG row card form and multiple strip form detection card
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (8)
1. A novel coronavirus IgM-IgA-IgG antibody colloidal gold detection kit is characterized in that: respectively putting card strips for detecting IgM antibody, IgA antibody and IgG antibody into a card shell in parallel to assemble a detection card; the gold-labeled areas of the coated plates of the three card strips are respectively coated with colloidal gold-labeled anti-human IgM, colloidal gold-labeled anti-human IgA and colloidal gold-labeled anti-human IgG, the detection areas are coated with novel coronavirus recombinant antigens, and the quality control areas are coated with goat anti-mouse IgG; wherein the novel coronavirus recombinant antigen is S-RBD protein.
2. The novel coronavirus IgM-IgA-IgG antibody colloidal gold detection kit according to claim 1, characterized in that: the preparation method of the colloidal gold labeled anti-human IgA, the colloidal gold labeled anti-human IgM and the colloidal gold labeled anti-human IgG comprises the following steps:
adding K to homogeneous colloidal gold solution2CO3Adjusting the pH to 6.0-10.0, adjusting the pH to 6.5-8.0 when coupling an antibody, adjusting the pH to 8.0-10.0 when coupling an antigen, adding anti-human IgA to the solution with the adjusted pH to ensure that the protein content of the anti-human IgA is 15-60 mu g/mL, coupling for 0.5-4 hours, adding a stop solution to the solution to stop the coupling reaction, wherein the final concentration of the stop solution is 1-1.5%; centrifuging after 0.5-2 hours after the reaction is ended, centrifuging for 15-30min at the centrifugal parameter of 12000-18000RPM, discarding the supernatant, redissolving the precipitate by using a redissolution, and storing at 2-8 ℃ for later use; the IgM and IgG antibodies are labeled by the same method as IgA.
3. A novel coronavirus IgM-IgA-IgG antibody colloidal gold detection kit is characterized in that: the detection card comprises a coating plate and a sample pad, wherein colloidal gold particles in a gold mark area of the coating plate are coated with a novel coronavirus recombinant antigen and biotinylated bovine serum albumin, different positions of the detection area are respectively coated with anti-human IgM, anti-human IgA and anti-human IgG, and a quality control area is coated with streptavidin; wherein the novel coronavirus recombinant antigen is S-RBD protein.
4. The novel coronavirus IgM-IgA-IgG antibody colloidal gold detection kit according to claim 3, wherein: preparation method of novel coronavirus recombinant antigen coated with colloidal gold particles comprises coating colloidal gold solution with 0.1M K2CO3Adjusting pH to 8.5-9.0, mixing thoroughly for 5-10 min, adding novel coronavirus recombinant antigen to the mixture to a final concentration of 10-30 μ g/mL, adding bovine serum albumin to the mixture to a final concentration of 0.8-1.2% after 10-30 min, discarding the supernatant after 30-60 min, 10000RPM, centrifuging for 10-15min, discarding the supernatant, and redissolving the precipitate with a redissolution; spraying the colloidal gold conjugate on a glass fiber membrane according to the spraying amount of 45-60 muL/cm, and drying at 35-45 ℃.
5. The novel coronavirus IgM-IgA-IgG antibody colloidal gold detection kit according to any one of claims 1 to 4, wherein: the sample pad is coated with sample treatment liquid, and the sample treatment liquid is tris buffer solution which contains BSA with the mass concentration of 0.5-1.5%, casein with the mass concentration of 0.5-1%, sucrose with the mass concentration of 0.3-1.2%, trehalose with the mass concentration of 0.1-1.2%, Procline with the volume concentration of 0.05-0.3%, tween-20 with the volume concentration of 0.05-0.5% and has the pH value of 7.0-7.6; the preparation method of the sample pad comprises spraying or soaking the sample pad treating solution onto or into the glass fiber film or polyester film at 45-85 microliter per square centimeter, and oven drying at 37-45 deg.C for 8-16 hr.
6. The novel coronavirus IgM-IgA-IgG antibody colloidal gold detection kit according to any one of claims 1 to 4, wherein: the colloidal gold solution is prepared by preparing chloroauric acid into a solution with the mass volume ratio of 1-10%, adding the solution with the final concentration of one ten thousandth to four ten thousandth into boiled purified water, continuously boiling for 2-5 minutes, adding 500-800 microliter of sodium citrate with the concentration of 0.1-0.2M into each 100mL of chloroauric acid solution, and continuously stirring and heating for 10-20 minutes to prepare the colloidal gold solution with uniform particle size.
7. The novel coronavirus IgM-IgA-IgG antibody colloidal gold detection kit according to any one of claims 1 to 4, wherein: also comprises sample diluent, 0.05M PB buffer solution with pH of 8.0, and NaCl with mass-volume ratio of 0.5-3% and sucrose with mass-volume ratio of 0.5-5%.
8. Use of a kit according to any one of claims 1 to 7 for the preparation of a novel reagent for the detection of coronavirus IgM-IgA-IgG antibodies.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113341138A (en) * | 2021-05-21 | 2021-09-03 | 长沙兴嘉生物工程股份有限公司 | Combined detection card for simultaneously detecting ASFV specific IgM and IgG antibodies and preparation method and application thereof |
| CN113834934A (en) * | 2021-11-29 | 2021-12-24 | 北京百普赛斯生物科技股份有限公司 | Immunochromatography test strip and preparation method thereof |
| CN114252601A (en) * | 2021-11-08 | 2022-03-29 | 润和生物医药科技(汕头)有限公司 | New corona total antibody and neutralizing antibody latex rapid detection test paper and preparation method thereof |
Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004109289A1 (en) * | 2003-06-10 | 2004-12-16 | Agency For Science, Technology And Research | Method of diagnosing sars corona virus infection |
| CN106198976A (en) * | 2016-06-30 | 2016-12-07 | 厦门宝太生物科技有限公司 | A kind of for detecting the reagent card of pepsin concn, test kit and purposes |
| CN106841601A (en) * | 2016-12-30 | 2017-06-13 | 广州市达瑞生物技术股份有限公司 | The preparation of zika virus multiple clips fusion protein and IgG/IgM antibody assay kits |
| CN109112219A (en) * | 2018-09-14 | 2019-01-01 | 赵风源 | Quickly detect the kit of forbidden amphetamine in entry passenger's belongings |
| CN109187986A (en) * | 2018-08-21 | 2019-01-11 | 施康培医疗科技（武汉）有限公司 | A kind of test strips and preparation method thereof of portable inspectiont people anti-thyroglobulin antibody |
| CN111089962A (en) * | 2020-03-25 | 2020-05-01 | 中山生物工程有限公司 | Colloidal gold kit for joint detection of novel coronavirus IgM/IgG antibody and preparation method thereof |
| CN111233985A (en) * | 2020-02-09 | 2020-06-05 | 北京丹大生物技术有限公司 | Preparation method of novel coronavirus IgA antibody rapid detection test strip |
| CN111337689A (en) * | 2020-04-03 | 2020-06-26 | 山西医科大学 | Novel coronavirus detection kit |
| CN111413495A (en) * | 2020-05-18 | 2020-07-14 | 博奥赛斯（天津）生物科技有限公司 | Novel coronavirus IgM/IgG colloidal gold detection kit |
| CN111426840A (en) * | 2020-06-11 | 2020-07-17 | 北京中检安泰诊断科技有限公司 | Novel coronavirus detection test strip and preparation method and application thereof |
| CN111551712A (en) * | 2020-05-07 | 2020-08-18 | 江苏苏博生物医学科技南京有限公司 | Novel test strip and kit for quickly detecting coronavirus IgM/IgG two-in-one and preparation method of test strip and kit |
| CN111562365A (en) * | 2020-05-29 | 2020-08-21 | 浙江省人民医院 | Novel coronavirus IgG/IgA antibody detection kit |
| CN111638332A (en) * | 2020-07-13 | 2020-09-08 | 浙江省人民医院 | Novel coronavirus IgA/IgM/IgG antibody ELISA detection kit |
-
2020
- 2020-09-11 CN CN202010955263.3A patent/CN112114139A/en active Pending
Patent Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004109289A1 (en) * | 2003-06-10 | 2004-12-16 | Agency For Science, Technology And Research | Method of diagnosing sars corona virus infection |
| CN106198976A (en) * | 2016-06-30 | 2016-12-07 | 厦门宝太生物科技有限公司 | A kind of for detecting the reagent card of pepsin concn, test kit and purposes |
| CN106841601A (en) * | 2016-12-30 | 2017-06-13 | 广州市达瑞生物技术股份有限公司 | The preparation of zika virus multiple clips fusion protein and IgG/IgM antibody assay kits |
| CN109187986A (en) * | 2018-08-21 | 2019-01-11 | 施康培医疗科技（武汉）有限公司 | A kind of test strips and preparation method thereof of portable inspectiont people anti-thyroglobulin antibody |
| CN109112219A (en) * | 2018-09-14 | 2019-01-01 | 赵风源 | Quickly detect the kit of forbidden amphetamine in entry passenger's belongings |
| CN111233985A (en) * | 2020-02-09 | 2020-06-05 | 北京丹大生物技术有限公司 | Preparation method of novel coronavirus IgA antibody rapid detection test strip |
| CN111089962A (en) * | 2020-03-25 | 2020-05-01 | 中山生物工程有限公司 | Colloidal gold kit for joint detection of novel coronavirus IgM/IgG antibody and preparation method thereof |
| CN111337689A (en) * | 2020-04-03 | 2020-06-26 | 山西医科大学 | Novel coronavirus detection kit |
| CN111551712A (en) * | 2020-05-07 | 2020-08-18 | 江苏苏博生物医学科技南京有限公司 | Novel test strip and kit for quickly detecting coronavirus IgM/IgG two-in-one and preparation method of test strip and kit |
| CN111413495A (en) * | 2020-05-18 | 2020-07-14 | 博奥赛斯（天津）生物科技有限公司 | Novel coronavirus IgM/IgG colloidal gold detection kit |
| CN111562365A (en) * | 2020-05-29 | 2020-08-21 | 浙江省人民医院 | Novel coronavirus IgG/IgA antibody detection kit |
| CN111426840A (en) * | 2020-06-11 | 2020-07-17 | 北京中检安泰诊断科技有限公司 | Novel coronavirus detection test strip and preparation method and application thereof |
| CN111638332A (en) * | 2020-07-13 | 2020-09-08 | 浙江省人民医院 | Novel coronavirus IgA/IgM/IgG antibody ELISA detection kit |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113341138A (en) * | 2021-05-21 | 2021-09-03 | 长沙兴嘉生物工程股份有限公司 | Combined detection card for simultaneously detecting ASFV specific IgM and IgG antibodies and preparation method and application thereof |
| CN114252601A (en) * | 2021-11-08 | 2022-03-29 | 润和生物医药科技(汕头)有限公司 | New corona total antibody and neutralizing antibody latex rapid detection test paper and preparation method thereof |
| CN113834934A (en) * | 2021-11-29 | 2021-12-24 | 北京百普赛斯生物科技股份有限公司 | Immunochromatography test strip and preparation method thereof |
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