CN110954696A - Aspergillus galactomannan detection kit and application thereof - Google Patents
Aspergillus galactomannan detection kit and application thereofDownload PDFInfo
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Abstract
Description
Technical Field
The invention relates to the technical field of aspergillus galactomannan detection, in particular to a aspergillus galactomannan detection kit and application thereof.
Background
For the current market of invasive fungus in vitro diagnosis, diagnostic products are mainly represented by ELISA detection kits. Taking the Aspergillus galactomannan detection product of BioRad as an example, the kit adopts a sandwich method, and the detection sample can only be serum of a patient. In the detection process, a treatment solution is added into a sample and pretreatment steps such as high-temperature heat treatment, centrifugation and the like are required. And then, mixing the processed sample and the enzyme-labeled antibody, adding the mixture into a micropore plate coated with the capture antibody in advance, incubating for 1.5h, washing the plate for 5 times, adding a substrate for color development for 30min, and detecting and reading after the reaction is stopped. The reading value is positively correlated with the concentration of the substance to be detected in the sample. The total time of the detection method is more than 3h, and the defects of troublesome pretreatment and low sensitivity exist, in addition, the method adopts the same IgM antibody to carry out a sandwich method, the stability of the kit is poor, and the kit is a qualitative detection product; secondly, the method carries out result interpretation according to the ratio of the detected OD values, and the detection range is small.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide an aspergillus galactomannan detection kit which has the advantages of no pretreatment of a detection sample, short detection time, high sensitivity and quantitative determination and application thereof.
The purpose of the invention is realized by the following technical scheme:
the utility model provides a aspergillus galactomannan detect reagent box, includes and is equipped with sample pad, colloidal gold pad, cellulose nitrate membrane and absorbent pad in proper order by detecting sample infiltration direction, the colloidal gold is filled up and is had the fluorescence microballon of the specific antibody of coupling anti aspergillus, the specific antibody detection section of anti aspergillus has on the cellulose nitrate membrane. The detection kit comprises a sample pad, a colloidal gold pad, an anti-aspergillus galactomannan antibody detection section, a nitrocellulose membrane, a mouse monoclonal antibody detection section and an anti-aspergillus galactomannan antibody detection section, wherein the sample pad is used for absorbing impurities and filtering an aspergillus galactomannan detection sample, the colloidal gold pad is used for combining the aspergillus galactomannan with fluorescent microspheres coupled with anti-aspergillus specific antibodies to form antibody antigen immune complexes, the nitrocellulose membrane receives the antibody antigen immune complexes on the colloidal gold pad and the fluorescent microspheres coupled with mouse monoclonal antibodies on the colloidal gold pad, the antibody antigen immune complexes are combined with the anti-aspergillus specific antibody detection section on the nitrocellulose membrane, and the fluorescent microspheres coupled with the mouse monoclonal antibodies are combined with the specific antibody detection section of; the immune complex is used for judging whether the fluorescent microspheres coupled with the mouse monoclonal antibody are combined with the detection section of the anti-aspergillus specific antibody or not, and when the water absorption pad is used for absorbing redundant detection samples.
Specifically, the downstream end of the sample pad is lapped on the colloidal gold pad, and the downstream end of the colloidal gold pad and the upstream end of the water absorption pad are lapped on the nitrocellulose membrane. The downstream of the sample pad is lapped on the colloidal gold pad, so that the sample can be absorbed and drained more quickly and effectively to be detected. The sample pad begins the infiltration according to the detection sample infiltration direction and is sample pad upstream end, what permeate at last is the downstream end, colloidal gold pad and absorbent pad also are the same reason, with sample pad downstream end overlap joint the infiltration and the filtration that are favorable to the sample pad on the colloidal gold pad more, colloidal gold pad downstream end overlap joint is favorable to the nitrocellulose membrane to adsorb the antibody antigen immune complex on the colloidal gold pad and the fluorescence microballon of coupling mouse monoclonal antibody more on the nitrocellulose membrane, accelerate detection speed, when detecting the sample too much through absorbent pad overlap joint can be quick absorption unnecessary detection sample on upstream end overlap joint nitrocellulose membrane more.
Specifically, still include the bottom plate, sample pad, colloidal gold pad, nitrocellulose membrane and absorbent pad set gradually on the bottom plate from left to right, and above-mentioned structure detects the sample and permeates usually from left to right.
Specifically, still include the bottom plate, sample pad, colloidal gold pad, nitrocellulose membrane and absorbent pad set gradually on the bottom plate from the center to around, and above-mentioned detection structure is by the center to infiltration all around.
Specifically, the bottom plate is arranged from the sample pad placing area to the absorbent pad placing area in an inclined and downward manner. The detection sample can be quickly permeated into the nitrocellulose membrane area through the inclined arrangement, and the detection rate is improved.
Specifically, the sample pad is completely superimposed on the colloidal gold pad. Further receive the effect of gravity and can be filtered the entering colloidal gold pad district by the sample pad completely when detecting the sample and need get into the colloidal gold pad, reach abundant filterable effect.
Specifically, the colloidal gold pad is further provided with fluorescent microspheres coupled with a mouse monoclonal antibody, the cellulose nitrate film pasting region is further provided with a specific antibody detection section for resisting mouse IgG, and the specific antibody detection section for resisting mouse IgG is closer to the water absorption pad than the specific antibody detection section for resisting aspergillus. The fluorescent microspheres coupled with the mouse monoclonal antibody and the antibody-antigen immune complex combined by the aspergillus galactomannan and the fluorescent microspheres coupled with the anti-aspergillus specific antibody enter a nitrocellulose membrane pasting region together, the antibody-antigen immune complex combined by the aspergillus galactomannan and the fluorescent microspheres coupled with the anti-aspergillus specific antibody is combined with an anti-aspergillus specific antibody detection section to be used as an experimental group, the fluorescent microspheres coupled with the mouse monoclonal antibody and a specific antibody detection section of anti-mouse IgG are combined to be used as a control group of the experimental group, when the content of the aspergillus galactomannan is detected to be lower, if the control group does not form the immune complex, the content of the aspergillus galactomannan in a detected sample is less or the sample does not enter a nitrocellulose membrane region, the detection result is inaccurate, and when the control group forms the immune complex, the content of the detected sample is up to the standard, the result is accurate.
The conjugated mouse monoclonal antibody is prepared by the following method:
1) buffer solution replacement for microspheres: adding 100 mu L of unloaded microspheres into a 1.5mL EP tube, adding 400 mu LMES buffer solution, uniformly mixing, centrifuging at the temperature of 10 ℃ and the rotating speed of 20000-30000 g for 10-20 min, discarding supernatant, adding 500 mu LMES buffer solution into the microsphere precipitate respectively, blowing and sucking by a pipette, uniformly mixing, and performing ultrasonic dispersion for 3-5 min by using an ultrasonic crusher;
2) activation of microspheres: the microsphere solution is added with 200-300 mu g of EDC dropwise while oscillating, 5-10 mu L of 10mg/mL Sulfo-NHS solution is added dropwise while oscillating, the solution is wrapped by tinfoil paper and protected from light, and the solution is activated for 15-20min at 40r/min of a room temperature rotary instrument;
3) washing the microspheres: centrifuging at 20000-30000 g for 10-20 min at 10 ℃, discarding the supernatant, adding 500 μ L HEPES buffer solution into the microsphere precipitate, blowing and sucking by a pipettor, and ultrasonically dispersing for 3-5 min by using an ultrasonic crusher;
4) marking the microspheres: adding 0.1mg antibody dropwise while oscillating the microsphere solution, wrapping with tin foil paper in a dark place, and reacting at room temperature with a rotary instrument at 40-60r/min for 120 +/-20 min;
5) sealing the microspheres: adding 55 μ L of sealing liquid into the microspheres, wrapping with tinfoil paper in dark place, and reacting at room temperature with a rotary instrument at 40-60r/min for 120 + -20 min;
6) washing the microspheres: centrifuging at 20000-30000 g for 10-20 min at 10 ℃, discarding the supernatant, adding 1mL of HEPES buffer solution into the microsphere precipitate, and blowing and sucking by a pipettor to mix uniformly. Repeating the washing operation for two times; the final pellet was resuspended in 200. mu.L of the conjugate stock. And ultrasonically dispersing for 3-5 min by using an ultrasonic crusher, and placing in a refrigerator at 4 ℃ to keep out of the sun to finish the preparation.
The application of the aspergillus galactomannan detection kit in the detection of the aspergillus galactomannan comprises the following steps:
1) filtering a sample solution containing aspergillus galactomannan, and combining the sample solution with fluorescent microspheres coupled with anti-aspergillus specific antibodies to form antibody-antigen immune complexes;
2) combining the antibody-antigen immune complex with the anti-aspergillus specific antibody detection section to form a final product;
3) and (3) measuring the fluorescence intensity of the final product by using a time-resolved fluorescence analyzer, and estimating the concentration of the aspergillus galactomannan in the reaction system according to the ratio of the fluorescence intensity to the relative fluorescence intensity.
Specifically, in step 2), a fluorescent microsphere coupled with a mouse monoclonal antibody is combined with a specific antibody detection segment of the anti-mouse IgG.
Compared with the prior art, the invention has the following advantages and beneficial effects:
(1) according to the invention, the sample pad is arranged to absorb impurities and filter aspergillus galactomannan, so that a sample to be detected is free from complex pretreatment, and the sample can be directly detected; the fluorescent microspheres coupled with the anti-aspergillus specific antibody are combined with aspergillus galactomannan to carry out detection, so that the detection range of a sample is greatly expanded, and the product can be used for detecting serum, plasma, cerebrospinal fluid, urine, pleural effusion, alveolar lavage fluid and the like.
(2) The invention detects the aspergillus galactomannan by the principle of time-resolved immunofluorescence chromatography, and compared with a sandwich method, the detection time is shorter and the sensitivity is higher. According to the invention, two specific mouse monoclonal antibodies are adopted for reaction, the affinity of the antibodies is high, the condition that single antibodies compete with each other is eliminated, and the detection can be realized without pretreatment. The detection can be realized within 10min, and the product detection time is greatly shortened. The detection kit for the aspergillus galactomannan can realize quantitative detection, and obviously overcomes the defects that the traditional colloidal gold method and ELISA method cannot carry out quantitative detection on the aspergillus galactomannan in a detected sample or the quantitative detection process is complex in operation, needs a pretreatment process, and has large detection result error and long time consumption.
(3) According to the invention, the colloidal gold pad is provided with the fluorescent microspheres coupled with the anti-aspergillus specific antibody and the fluorescent microspheres coupled with the mouse monoclonal antibody, the nitrocellulose membrane is provided with the anti-aspergillus specific antibody detection section and the anti-mouse IgG specific antibody detection section, an antibody antigen immune complex combined by the aspergillus galactomannan and the fluorescent microspheres coupled with the anti-aspergillus specific antibody is combined with the anti-aspergillus specific antibody detection section to be used as a detection group, and the fluorescent microspheres coupled with the mouse monoclonal antibody and the anti-mouse IgG specific antibody detection section are combined to be used as a control group, so that the stability and the repeatability are better, and the result error is obviously reduced.
Drawings
FIG. 1 is a schematic diagram of a kit for detecting Aspergillus galactomannan.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
As shown in figure 1, the aspergillus galactomannan detection kit comprises asample pad 2, acolloidal gold pad 3, a nitrocellulose membrane 4 and awater absorption pad 5 which are sequentially arranged in the permeation direction of a detection sample, wherein thecolloidal gold pad 3 is provided with fluorescent microspheres coupled with an anti-aspergillus specific antibody, and the nitrocellulose membrane 4 is provided with an anti-aspergillus specificantibody detection section 6. Compared with the prior ELISA detection technology, because the used antibody has low affinity, the detection kit needs to carry out high-temperature pretreatment, centrifugation and other steps and needs to wait for reaction for two hours during detection, the detection kit detects the content of galactomannan released to the outside by the growth of fungi such as aspergillus and the like through a time-resolved immunofluorescence chromatography method, so that the detection kit can only detect the serum of a patient compared with the traditional ELISA method, the detection kit can only detect the serum of the patient through the serum, the plasma, the cerebrospinal fluid, the urine, the pleural effusion, the alveolar lavage fluid and other liquids of the patient, the applicable sample range is greatly increased, two specific mouse monoclonal antibodies are adopted for reaction, the antibody affinity is high, and the condition that single antibodies compete with each other is eliminated, detection can be realized without pretreatment. Thesample pad 2 is preferably a glass cellulose membrane, the material of thecolloidal gold pad 3 is preferably colloidal gold, the nitrocellulose membrane 4 is preferably a nitrocellulose membrane 4, and theabsorbent pad 5 is a conventionalabsorbent pad 5. The fluorescence microsphere coupled with the aspergillus specific antibody is provided with lanthanide, fluorescence is detected by a time-division technology through the luminescence characteristic of lanthanide chelate, and two parameters of detection wavelength and time are simultaneously used for signal resolution.
Specifically, the downstream end of thesample pad 2 is lapped on thecolloidal gold pad 3, and the downstream end of thecolloidal gold pad 3 and the upstream end of thewater absorption pad 5 are lapped on the nitrocellulose membrane 4.
Specifically, still include bottom plate 1,sample pad 2,colloidal gold pad 3, nitrocellulose membrane 4 andabsorbent pad 5 set gradually on bottom plate 1 from left to right.
Specifically, the device further comprises a bottom plate 1, and thesample pad 2, thecolloidal gold pad 3, the nitrocellulose membrane 4 and theabsorbent pad 5 are sequentially arranged on the bottom plate 1 from the center to the periphery.
Specifically, the bottom plate 1 is arranged from thesample pad 2 placing area to theabsorbent pad 5 placing area in an inclined and downward manner.
Specifically, thesample pad 2 is completely superimposed on thecolloidal gold pad 3.
Specifically, thecolloidal gold pad 3 is further provided with fluorescent microspheres coupled with a mouse monoclonal antibody, the nitrocellulose membrane 4 is further provided with a specificantibody detection section 7 for resisting mouse IgG on the pasting region, and the specificantibody detection section 7 for resisting mouse IgG is closer to thewater absorption pad 5 than the specificantibody detection section 6 for resisting aspergillus. The anti-aspergillus specificantibody detection section 6 and the anti-mouse IgG specificantibody detection section 7 are two different mouse monoclonal antibodies, and the two monoclonal antibodies are different in the corresponding sites, so that the product stability is strong, and the batch difference is small. Compared with the traditional ELISA method, the method has the advantages that competition and steric hindrance caused by only one antibody are avoided, and the product is poor in stability and large in batch-to-batch difference.
The conjugated mouse monoclonal antibody is prepared by the following method:
1) buffer solution replacement for microspheres: adding 100 mu L of unloaded microspheres into a 1.5mL EP tube, adding 400 mu LMES buffer solution, uniformly mixing, centrifuging at the temperature of 10 ℃ and the rotating speed of 20000-30000 g for 10-20 min, discarding supernatant, adding 500 mu LMES buffer solution into the microsphere precipitate respectively, blowing and sucking by a pipette, uniformly mixing, and performing ultrasonic dispersion for 3-5 min by using an ultrasonic crusher;
2) activation of microspheres: the microsphere solution is added with 200-300 mu g of EDC dropwise while oscillating, 5-10 mu L of 10mg/mL Sulfo-NHS solution is added dropwise while oscillating, the solution is wrapped by tinfoil paper and protected from light, and the solution is activated for 15-20min at 40r/min of a room temperature rotary instrument;
3) washing the microspheres: centrifuging at 20000-30000 g for 10-20 min at 10 ℃, discarding the supernatant, adding 500 μ L HEPES buffer solution into the microsphere precipitate, blowing and sucking by a pipettor, and ultrasonically dispersing for 3-5 min by using an ultrasonic crusher;
4) marking the microspheres: adding 0.1mg antibody dropwise while oscillating the microsphere solution, wrapping with tin foil paper in a dark place, and reacting at room temperature with a rotary instrument at 40-60r/min for 120 +/-20 min;
5) sealing the microspheres: adding 55 μ L of sealing liquid into the microspheres, wrapping with tinfoil paper in dark place, and reacting at room temperature with a rotary instrument at 40-60r/min for 120 + -20 min;
6) washing the microspheres: centrifuging at 20000-30000 g for 10-20 min at 10 ℃, discarding the supernatant, adding 1mL of HEPES buffer solution into the microsphere precipitate, and blowing and sucking by a pipettor to mix uniformly. Repeating the washing operation for two times; the final pellet was resuspended in 200. mu.L of the conjugate stock. And ultrasonically dispersing for 3-5 min by using an ultrasonic crusher, and placing in a refrigerator at 4 ℃ to keep out of the sun to finish the preparation. Wherein, the ultrasonic crusher in the step 1, thestep 3 and thestep 6 should carry out ice bath on the sample when in work, the working state is adjusted to 10 percent of power, and the amplitude transformer
Working for 2s, stopping for 2s, and reciprocating.
The preparation method of the colloidal gold pad comprises the following steps of adding fluorescent microspheres coupled with an anti-aspergillus specific antibody and fluorescent microspheres coupled with a mouse monoclonal antibody into the colloidal gold pad to form a marking pad, specifically, preparing the marking pad by adopting the following steps, preparing a fluorescent microsphere-labeled anti-aspergillus galactomannan IgG antibody turbid liquid by adopting a coupled mouse monoclonal antibody preparation method, using the turbid liquid for marking the pad, spraying the turbid liquid on the marking pad, wherein the spraying amount is 25-35 mu L/cm, after the spraying is finished, placing the marking pad in an oven with the humidity of less than 20% at 40-50 ℃, drying for 12-24 h, and then sealing and storing at 2-30 ℃.
The nitrocellulose membrane is introduced with an anti-aspergillus specific antibody detection section and an anti-mouse IgG specific antibody detection section to form a chromatographic pad, and the chromatographic pad is prepared by the following steps: adjusting the concentration of the aspergillus fumigatus mannan antibody to be 0.5-1 mg/mL by using a coating buffer solution, and then respectively spraying the aspergillus fumigatus mannan antibody onto a nitrocellulose membrane, wherein the spraying amount is 0.1-0.2 muL/mm, and the T line is obtained. Adjusting the concentration of goat anti-mouse IgG to 0.5-1 mg/mL by using a coating buffer solution, and then respectively spraying the goat anti-mouse IgG onto a nitrocellulose membrane, wherein the spraying amount is 0.1-0.2 muL/mm, and the C line is obtained. And after the spraying is finished, placing the nitrocellulose membrane in a 40-50 ℃ oven with the humidity less than 20%, drying for 24-72 h, and then sealing and storing at 2-30 ℃ to finish the preparation.
Specifically, the sample pad was treated using the following method: and soaking the sample pad in a sealing solution, placing the sample pad in an oven with the humidity of less than 20% and the temperature of 40-50 ℃, drying for 12-24 h, and then sealing and storing at 2-30 ℃.
The application of the aspergillus galactomannan detection kit in the detection of the aspergillus galactomannan comprises the following steps:
1) filtering a sample solution containing aspergillus galactomannan, and combining the sample solution with fluorescent microspheres coupled with anti-aspergillus specific antibodies to form antibody-antigen immune complexes;
2) combining the antibody-antigen immune complex with thedetection section 6 of the anti-aspergillus specific antibody to form a final product;
3) and (3) measuring the fluorescence intensity of the final product by using a time-resolved fluorescence analyzer, and estimating the concentration of the aspergillus galactomannan in the reaction system according to the ratio of the fluorescence intensity to the relative fluorescence intensity.
Specifically, in step 2), a fluorescent microsphere coupled with a mouse monoclonal antibody is combined with theantibody detection section 7 specific to the mouse IgG.
The specific implementation process of the invention is as follows:
dripping a detection sample on thesample pad 2, standing for 10min, measuring fluorescence by a time resolution technology, simultaneously detecting two parameters of wavelength and time to perform signal resolution, and finally determining the concentration value of galactomannan to further determine the content of aspergillus.
Compared with a colloidal gold method and an ELISA method, the method can realize quantitative detection, obviously overcomes the defects that the traditional colloidal gold method and the ELISA method cannot carry out quantitative detection on the galactomannan in a detection sample or the quantitative detection process is complex in operation, needs a pretreatment process, has large detection result error and consumes long time, can realize high-sensitivity detection, and has the detection sensitivity of 10-9mol/L, detection sensitivity of radioimmunoassay 10-12mol/L, chemiluminescence method detection sensitivity of10-15mol/L. The time-resolved fluorescence method adopted in the patent has the detection sensitivity of 10-18mol/L, much higher than other assays.
The above-mentioned embodiments only express the embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A aspergillus galactomannan detect reagent box, its characteristic lies in: include and be equipped with sample pad, colloidal gold pad, cellulose nitrate membrane and the pad that absorbs water in proper order by detecting sample infiltration direction, the colloidal gold pads up has the fluorescence microballon of the anti aspergillus specific antibody of coupling, the last anti aspergillus specific antibody detection section that has of cellulose nitrate membrane.
2. The aspergillus galactomannan detection kit according to claim 1, wherein: the downstream end of the sample pad is lapped on the colloidal gold pad, and the downstream end of the colloidal gold pad and the upstream end of the water absorption pad are lapped on the nitrocellulose membrane.
3. The aspergillus galactomannan detection kit according to claim 2, wherein: still include the bottom plate, sample pad, colloidal gold pad, nitrocellulose membrane and absorbent pad set gradually on the bottom plate from left to right.
4. The aspergillus galactomannan detection kit according to claim 2, wherein: the sample pad, the colloidal gold pad, the nitrocellulose membrane and the absorbent pad are sequentially arranged on the bottom plate from the center to the periphery.
5. The Aspergillus galactomannan detection kit according to claim 3 or 4, wherein: the bottom plate is obliquely and downwards arranged from the sample pad placing area to the water absorption pad placing area.
6. The Aspergillus galactomannan detection kit according to claim 3 or 4, wherein: the sample pad is completely superposed on the colloidal gold pad.
7. The Aspergillus galactomannan detection kit according to any one of claims 1 to 4, wherein: the colloidal gold pad is also provided with fluorescent microspheres coupled with a mouse monoclonal antibody, the cellulose nitrate film pasting region is also provided with a specific antibody detection section for resisting mouse IgG, and the specific antibody detection section for resisting mouse IgG is closer to the water absorption pad than the specific antibody detection section for resisting aspergillus.
8. The aspergillus galactomannan detection kit according to claim 7, wherein: the conjugated mouse monoclonal antibody is prepared by the following method:
1) buffer solution replacement for microspheres: adding 100 mu L of unloaded microspheres into a 1.5mL EP tube, adding 400 mu LMES buffer solution, uniformly mixing, centrifuging at the temperature of 10 ℃ and the rotating speed of 20000-30000 g for 10-20 min, discarding supernatant, adding 500 mu L MES buffer solution into the microsphere precipitate respectively, blowing and sucking by a pipette, uniformly mixing, and performing ultrasonic dispersion for 3-5 min by using an ultrasonic crusher;
2) activation of microspheres: the microsphere solution is added with 200-300 mu g of EDC dropwise while oscillating, 5-10 mu L of 10mg/mL Sulfo-NHS solution is added dropwise while oscillating, the solution is wrapped by tinfoil paper and protected from light, and the solution is activated for 15-20min at 40r/min of a room temperature rotary instrument;
3) washing the microspheres: centrifuging at 20000-30000 g for 10-20 min at 10 ℃, discarding the supernatant, adding 500 μ L HEPES buffer solution into the microsphere precipitate, blowing and sucking by a pipettor, and ultrasonically dispersing for 3-5 min by using an ultrasonic crusher;
4) marking the microspheres: adding 0.1mg antibody dropwise while oscillating the microsphere solution, wrapping with tin foil paper in a dark place, and reacting at room temperature with a rotary instrument at 40-60r/min for 120 +/-20 min;
5) sealing the microspheres: adding 55 μ L of sealing liquid into the microspheres, wrapping with tinfoil paper in dark place, and reacting at room temperature with a rotary instrument at 40-60r/min for 120 + -20 min;
6) washing the microspheres: centrifuging at 20000-30000 g for 10-20 min at 10 ℃, discarding the supernatant, adding 1mL of HEPES buffer solution into the microsphere precipitate, and blowing and sucking by a pipettor to mix uniformly. Repeating the washing operation for two times; the final pellet was resuspended in 200. mu.L of the conjugate stock. And ultrasonically dispersing for 3-5 min by using an ultrasonic crusher, and placing in a refrigerator at 4 ℃ to keep out of the sun to finish the preparation.
9. The use of an Aspergillus galactomannan detection kit according to claim 9 wherein in step 2) fluorescent microspheres conjugated with murine monoclonal antibodies are conjugated to an anti-mouse IgG specific antibody detection segment.
10. The use of an Aspergillus galactomannan detection kit according to claim 8 wherein in step 2) fluorescent microspheres conjugated with murine monoclonal antibodies are conjugated to an anti-mouse IgG specific antibody detection segment.
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| CN114544934A (en)* | 2022-02-21 | 2022-05-27 | 丹娜(天津)生物科技股份有限公司 | Aspergillus galactomannan detection test strip and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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