CN105158471A - Detection kit for human parvovirus B19 (HPVB19) type IgM (immunoglobulin) antibody and preparation method of detection kit - Google Patents
Detection kit for human parvovirus B19 (HPVB19) type IgM (immunoglobulin) antibody and preparation method of detection kit Download PDFInfo
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Abstract
The invention discloses a detection kit for a human parvovirus B19 (HPVB19) type IgM (immunoglobulin) antibody. The detection kit comprises a detection strip, wherein the detection strip comprises a gold conjugate pad and a reaction film, the gold conjugate pad is coated with a colloidal gold labeled mouse-anti-human IgM monoclonal antibody, the reaction film comprises a detection line and a quality control line, the detection line is coated with a recombinant HPVB19 antigen, and the quality control line is coated with a goat-anti-mouse IgG polyclonal antibody. The invention further discloses a preparation method of the kit. The preparation method comprises a reaction film preparation step, a gold conjugate pad preparation step as well as a cutting and assembly step. The kit is simple and convenient to operate, low in cost, good in specificity and high in sensitivity, is suitable for popularization and early-stage diagnosis, can perform single detection and is applicable to epidemiologic investigation and field application.
Description
Technical field
The present invention relates to field of immunodetection, refer to human parvovirus B19's type IgM antibody detection kit and preparation method thereof especially.
Background technology
Human parvovirus B19 type (HPVB19) is linear ssdna virus, and Genome Size is 5.6kb, classifies as the red Tobamovirus of parvovirus subfamily.The extraneous physics ability to function of this virus resistance is very strong, and virion is placed on 56 DEG C of 10min and can not be inactivated, and lipid solvent does not also affect it, in formalin, β-the third, may make its deactivation under cruel and gamma-ray effect.
Human parvovirus B19 type with blood product and the children in 2-7 year for major source of infection.It infects in worldwide distribution, and crowd's human parvovirus B19 type antibody positive rate accounts for 60-65%, and Area distribution is without significant difference.It is light-duty acute infection that human parvovirus B19 type infects general, if patient is with hematopoietic disorder, immunodeficiency or virus DNA integrates and host cell chromosome etc., can cause severe infections or the Chronic persistent course of disease, and harm humans is healthy.
At present, the main virulent separation of the detection of human parvovirus B19 type is cultivated, virion electron microscopic observation, the method such as serology and molecular biology.Wherein, Viral isolation detects the sample needed containing life virus, adds the possibility of infection.Further, need to cultivate in thermostatic equipment; In addition, incubation time is long, the incubation time usually needing at least 24 hours.And virion electron microscopic observation needs Electronic Speculum equipment, range of application is little, troublesome poeration, and the selection of observation point may cause undetected by interference from human factor in addition.Molecular Biological Detection needs the experimenter of professional equipment and specialty to complete detection, and testing process is loaded down with trivial details, and detection time is longer.Equally, in Serologic detection, radioimmunology (RIA), enzyme immunoassay (EIA), immunofluorescence technique (IF), Diagnosis of Sghistosomiasis notation (WB) and enzyme linked immunosorbent assay (ELISA) all need professional and operate, equipment is needed to carry out interpretation, detection time is all longer, in clinical practice, and the more difficult popularization in the aspect such as penetration and promotion, Site Detection.
Summary of the invention
In view of this, the object of the invention is to propose a kind of human parvovirus B19's type IgM antibody detection kit, easy and simple to handle, with low cost, specificity is good, highly sensitive, be suitable for popularizing, early diagnosis can single part of detection, be suitable for epidemiology survey and rig-site utilization.
Detector bar is comprised based on above-mentioned purpose human parvovirus B19's type provided by the invention IgM antibody detection kit, described detector bar comprises golden bond pad and reaction film, described golden bond pad is coated with the mouse-anti people IgM monoclonal antibody of colloid gold label, described reaction film comprises detection line and nature controlling line, described detection line is coated with restructuring HPVB19 antigen, and described nature controlling line is coated with sheep anti-mouse igg polyclonal antibody.
Preferably, described restructuring HPVB19 antigen is the VP1 albumen of human parvovirus B19's type of the recombinant expressed acquisition of using gene engineering method, and concentration is 2.0-10.0mg/mL.
Preferably, the PCR primer preparing described restructuring HPVB19 antigen used is:
5'-AGCCATATGAGTAAAGAAAGTGGCAAATGGT-3'；
5'-GCACTCGAGTTATGTACCTCCTGTACCTAAAAG-3'。
Optionally, described sheep anti-mouse igg Anti-TNF-α bulk concentration is 4.0-10.0mg/mL, and described mouse-anti people IgM monoclonal antibody concentration is 2.0-10.0mg/mL.
Optionally, also comprise Sample dilution, this Sample dilution is phosphate buffer.
Preferably, described Sample dilution pH value is 7.3-7.5, and conductivity is 8000 μ s/cm-12000 μ s/cm.
Preferably, described detector bar is positioned in rectangular plastic magazine, and form test card, this test card is provided with sample well and detect aperture, described sample well is corresponding with the described sample pad location of described detector bar, the described detection line of the corresponding described detector bar of described detect aperture and described nature controlling line.
Based on identical inventive concept, present invention also offers the preparation method of above-mentioned human parvovirus B19's type IgM antibody detection kit, comprise the following steps:
(1) preparation of reaction film:
Dilute described restructuring HPVB19 antigen and described sheep anti-mouse igg polyclonal antibody, and be sprayed on described reaction film, form described detection line and nature controlling line respectively, drying for standby;
(2) preparation of golden bond pad:
Described mouse-anti people IgM monoclonal antibody and collaurum are carried out the mouse-anti people IgM monoclonal antibody solution that coupling obtains colloid gold label, adjustment concentration, and soak described golden bond pad, paving gold, drying for standby;
(3) cutting assembling: described reaction film, golden bond pad, robust fibre filter paper, sample pad are assembled to described reaction plate, forms described detector bar after cutting.
Optionally, the preparation method of the mouse-anti people IgM monoclonal antibody of described colloid gold label comprises the following steps:
(1) solution of potassium carbonate is used to regulate collaurum pH value to 8.0-9.0;
(2) add mouse-anti people IgM monoclonal antibody, stir;
(3) add 10% bovine serum albumin solution, stir;
(4) centrifugal, abandoning supernatant;
(5) add colloidal gold conjugate dilution, to obtain final product.
Optionally, the preparation method of described collaurum boils for adding gold chloride in purified water, adds citric acid three sodium solution under agitation, boils, and treats that it cools naturally.
The principle of this kit is:
Adopt colloidal gold immunochromatographimethod know-why, the reorganized HPVB19 antigen of the detection line bag on nitrocellulose filter, at nature controlling line bag by sheep anti-mouse igg polyclonal antibody, gold mark pad wraps by the mouse-anti people IgM monoclonal antibody of colloid gold label.For the human parvovirus B19's type IgM antibody in qualitative detection whole blood or serum (slurry) sample.
When detecting positive sample, the human parvovirus B19's type IgM antibody in sample can be combined with the mouse-anti people IgM monoclonal antibody of colloid gold label, forms immune complex, because chromatography effect compound and sample are at the inner flow forward of nitrocellulose filter.When compound is combined with the restructuring HPVB19 antigen of bag quilt through detection line, form " collaurum-mouse-anti people IgM monoclonal antibody-human parvovirus B19's type IgM antibody-restructuring HPVB19 antigen " and aggegation develops the color.Residue sample is combined with the sheep anti-mouse igg polyclonal antibody of nature controlling line bag quilt and aggegation develops the color.When detecting negative sample, not containing human parvovirus B19's type IgM antibody in sample, cause and can not form immune complex, then can only develop the color at nature controlling line place.
As can be seen from above, human parvovirus B19's type IgM antibody detection kit provided by the invention can be used for early diagnosis, detect quick, easy, do not need specific installation, widely applicable, specificity is good, highly sensitive.The whole running time only needs 15-20 minute, is applicable to clinical detection, field investigation and epidemiology survey etc.
Accompanying drawing explanation
Fig. 1 is the structural representation of detector bar in embodiment of the present invention human parvovirus B19 type IgM antibody detection kit;
Fig. 2 is the structural representation of test card in embodiment of the present invention human parvovirus B19 type IgM antibody detection kit.
Embodiment
For making the object, technical solutions and advantages of the present invention clearly understand, below in conjunction with specific embodiment, and with reference to accompanying drawing, the present invention is described in more detail.
Embodiment one:
In the present embodiment, described human parvovirus B19's type IgM antibody detection kit, comprises detector bar or test card, Sample dilution.
Fig. 1 is the structural representation of detector bar in embodiment of the present invention human parvovirus B19 type IgM antibody detection kit.As shown in Figure 1, described detector bar comprises reaction plate 1, and adheres to the sample pad 2 of on described reaction plate 1, golden bond pad 3, reaction film 4, robust fibre filter paper 5.
Wherein, described golden bond pad 3 is coated with the mouse-anti people IgM monoclonal antibody of colloid gold label.Described mouse-anti people IgM monoclonal antibody concentration is 2.0-10.0mg/mL.Preferably, described mouse-anti people IgM monoclonal antibody concentration is 2mg/mL.This mouse-anti people IgM monoclonal antibody is achromaticity and clarification clear solution.Measure by polyacrylamide gel electrophoresis (SDS-PAGE), when applied sample amount is 10 μ L, this mouse-anti people IgM monoclonal antibody is shown as heavy chain and each band of light chain.Alternatively, described mouse-anti people IgM monoclonal antibody is purchased from Beijing ten thousand Yu Mei billows Science and Technology Ltd..
In the present embodiment, described reaction film 4 is nitrocellulose filter, comprises detection line 6 and nature controlling line 7.Described detection line 6 is coated with restructuring HPVB19 antigen, and described restructuring HPVB19 antigen concentration is 2.0-10.0mg/mL, achromaticity and clarification clear solution.Measure with SDS-PAGE or additive method, when applied sample amount is 10 μ L, only show a band.Preferably, described restructuring HPVB19 antigen is the VP1 albumen of human parvovirus B19's type virus.
Wherein, the preparation method of described restructuring HPVB19 antigen is:
(1) obtain the VP1 protein sequence information design antigen PCR primer of human parvovirus B19 according to GeneBank in NCBI, the DNA product extracting human parvovirus B19 from human parvovirus positive serum is template, carries out pcr amplification.PCR primer (comprising restriction enzyme site) is as follows:
Forward primer:
5'-AGCCATATGAGTAAAGAAAGTGGCAAATGGT-3'
Reverse primer:
5'-GCACTCGAGTTATGTACCTCCTGTACCTAAAAG-3'。
After pcr amplification, PCR primer 1.5% agarose gel electrophoresis is separated.Under the observation of gel imaging instrument, cut about 1365bp and locate object band.Adopt Ago-Gel to reclaim kit and purifying is carried out to object fragment.
(2) after reclaiming, PCR fragment is cloned in pMD18-T carrier, competent escherichia coli cell transforms, screening positive clone, PCR identifies, and is checked order, after order-checking is correct, enzyme is cut and is proceeded to expression vector pET28a, and transfection expression Host Strains e. coli bl21 (DE3) is expressed.
(3) recombination bacillus coli is cultivated, low temperature IPTG abduction delivering, collect the Escherichia coli of expressing, after ultrasonic disruption, adopt Ni post affinity chromatography method to obtain destination protein, measure with SDS-PAGE, when applied sample amount is 10 μ L, only 50KD shows a band.
In the present embodiment, described nature controlling line 7 is coated with sheep anti-mouse igg polyclonal antibody.The concentration of described sheep anti-mouse igg polyclonal antibody is 4.0-10.0mg/mL.Preferably, described sheep anti-mouse igg Anti-TNF-α bulk concentration is 4mg/mL.This sheep anti-mouse igg polyclonal antibody is achromaticity and clarification clear solution.Alternatively, described sheep anti-mouse igg polyclonal antibody is purchased from Beijing ten thousand Yu Mei billows Science and Technology Ltd..
In the present embodiment, described Sample dilution is 20mM, and pH value is the phosphate buffer of 7.4.Preferably, described Sample dilution is colourless, clarification, transparency liquid no suspended substance, and with PH measurement, determine pH value be 7.3-7.5, should be 8000 μ s/cm ~ 12000 μ s/cm by conductivity meter mensuration conductivity.
Fig. 2 is the structural representation of test card in embodiment of the present invention human parvovirus B19 type IgM antibody detection kit, as shown in Figure 2, as an optional embodiment, described detector bar is positioned in specific rectangular plastic magazine, form described test card, this test card is provided with a sample well 8 and detect aperture 9.Described sample well 8 is for dripping laboratory sample, corresponding with sample pad 2 position of described detector bar; The detection line 6 of the corresponding detector bar of detect aperture 9 and nature controlling line 7, can observe the change of detection line 6 and nature controlling line 7 on described reaction film 4 by detect aperture 9.
Embodiment two: the preparation method of human parvovirus B19's type IgM antibody detection kit
Utilize the antigen coated reaction film 4 of restructuring HPVB19 as detection line; Utilize sheep anti-mouse igg polyclonal antibody bag by reaction film 4 as nature controlling line; The mouse-anti people IgM monoclonal antibody bag of mark collaurum, by golden bond pad, is assembled and obtains detector bar.
(1) preparation of reaction film:
Being wrapped by concentration to the best by restructuring HPVB19 antigen diluent by dilution with bag is 1.0mg/mL-1.5mg/mL;
By dilution, sheep anti-mouse igg polyclonal antibody is diluted to best bag by concentration 1.5mg/mL-3.0mg/mL with bag;
Described bag is the phosphate buffer of 0.05M by dilution.Described bag is colourless, clarification, transparency liquid and no suspended substance by dilution.By the purified water twice dilution of this damping fluid, determining pH value with PH measurement is 7.3-7.5.By the purified water twice dilution of this damping fluid, measuring conductivity with conductivity meter is 9000-14000 μ s/cm.
Be sprayed onto on reaction film 4 with a film machine respectively by two kinds of coating buffers, specking amount is 0.05 μ L/mm-0.15 μ L/mm, forms described detection line 6 and nature controlling line 7 respectively; To wrap the reaction film 4 of quilt in 37 DEG C of dryings 3 hours, and in room temperature preservation.Described bag is the phosphate buffer of 0.05M by dilution.
As a preferred embodiment, specking amount is 0.1 μ L/mm, and nature controlling line bag is by concentration: sheep anti-mouse igg polyclonal antibody is 2.0mg/ml; Detection line bag is by concentration: restructuring HPVB19 antigen is 1.0mg/ml.When this concentration, reaction film nature controlling line and detection line color developing effect are better, and sensitivity, specificity is better.
(2) preparation of golden bond pad:
Mouse-anti people IgM monoclonal antibody and collaurum are carried out the mouse-anti people IgM monoclonal antibody solution that coupling obtains colloid gold label.
In the present embodiment, collaurum is prepared according to gold chloride under boiling conditions and trisodium citrate generation redox reaction.
In 100mL purified water, add 0.1mL10% gold chloride boil, add 1% citric acid three sodium solution 1.4-2.0mL under agitation, boil 5 minutes, the colloid gold particle of preparation, treat that it cools naturally and return back to original volume.Preferably, in 100mL purified water, add 0.1mL10% gold chloride boil, add 1.8mL1% citric acid three sodium solution under agitation, collaurum color prepared by this condition is scarlet, limpid, and without muddy and floating thing, its specificity of the collaurum after mark, susceptibility are better.
The mouse-anti people IgM monoclonal antibody solution of colloid gold label is adjusted to optium concentration 10-20 μ g/mL, with 1-2mL/ bar, soaks described golden bond pad, paving gold, 37 DEG C of dryings 3 hours, and in room temperature preservation.
Preferably, mouse-anti people IgM monoclonal antibody mark final concentration is when being 15 μ g/mL, with the specificity of 1.5mL/ bar test strips and susceptibility better, sensitivity is higher.
Wherein, the preparation process of the mouse-anti people IgM monoclonal antibody of colloid gold label is as follows:
1) regulate collaurum pH value to 8.0-9.0 with 0.1M solution of potassium carbonate;
2) add the mouse-anti people IgM monoclonal antibody of final concentration 15 μ g/mL, stir 40 minutes;
3) bovine serum albumin(BSA) to the final concentration adding 10% is again 1%, stirs 15 minutes;
4) 12000rpm/min, 4 DEG C are centrifugal 15 minutes, careful abandoning supernatant;
5) the colloidal gold conjugate dilution of 1/2 volume of original volume (original volume is with collaurum volume computing) is added.
Wherein, described colloidal gold conjugate dilution is dissolved in 1L deionized water sodium hydrogen phosphate 5.372g, sodium dihydrogen phosphate 0.78g, bovine serum albumin(BSA) 5g, sodium chloride 8.5g, casein 0.5g, and mixing is prepared from.
Preferably, step 1) middle with 0.1M solution of potassium carbonate adjustment collaurum pH value to 8.5, the effect of colloid gold label mouse-anti people IgM monoclonal antibody is better, good stability.
(3) cutting assembling: in interior parlor, reaction film, golden bond pad, robust fibre filter paper, sample pad are assembled into reaction plate, then are assembled into finished product with the detector bar that cutting machine is cut into 4mm wide.
Embodiment three: the detection method of human parvovirus B19's type IgM antibody detection kit
1. detector bar detection method
(1) from original packing aluminium foil bag, take out detector bar, be flat on table top;
(2) get 20 μ L serum or plasma samples, be added to the application of sample place of detector bar arrow lower end, then add Sample dilution 100 μ L (about 2-3 drips);
(3) sentence read result in 15-20 minute, after 20 minutes, assay is invalid.
(4) testing result judges:
Negative: only to occur a red stripes in nature controlling line position.
Positive: two red stripes appear in nature controlling line and detection line position.
Invalid: nature controlling line position does not occur red stripes
2. test card detection method
(1) from aluminium foil bag, take out test card, be placed on horizontal on horizontal operation table top, and carry out sample labeling;
(2) get 20 μ L serum, blood plasma or 40 μ l whole blood samples, directly join in well, then add Sample dilution 100 μ L (about 2-3 drips);
(3) sentence read result in 15-20 minute, after 20 minutes, assay is invalid.
(4) testing result judges:
Negative: only to occur a red stripes in nature controlling line position.
Positive: two red stripes appear in nature controlling line and detection line position.
Invalid: nature controlling line position does not occur red stripes.
Embodiment four: human parvovirus B19's type IgM antibody detection kit validation verification
The quality controlled serum adopting said method to prepare detects the validity of the present inventor's assays for parvovirus B 19 type IgM antibody detection kit:
(1) method of operating: be directly added in well by 10 μ L serum, then add 100 μ l Sample dilution, room temperature is placed, and 15 minutes sentence read result, may not exceed 20 minutes.
(2) result judges:
Positive: two red line appear in detection window, i.e. nature controlling line and detection line.
Negative: a red nature controlling line only appears in detection window.
Invalid: detection window redfree nature controlling line occurs.
(3) physical examination:
1) flat appearance, material adhesion-tight, content is complete.
2) random detection 10 test strips, liquid creep speed all >=10mm/min.
(4) susceptibility: 10 parts of enterprise reference material HPVB19-IgM Positive Sera P1-P10, colour developing result is all positive.
(5) specificity: 10 parts of enterprise reference material HPVB19-IgM negative antibody serum N 1-N10, colour developing result is all negative.
(6) minimum detectability: positive end-point should be not less than 1:8 and doubly dilute.
(7) accuracy: get enterprise's reference material accuracy serum to 10 ELISA test strips, its colour developing result is consistent, is the positive, and colour developing evenly.
(8) finished product kit storage and effect the phase: finished product kit through after the assay was approved, hand over warehouse for finished product management with storage, under being stored in 4-30 DEG C of condition, the term of validity is 12 months.
The human parvovirus B19's type IgM antibody detection kit that the present invention relates to has the following advantages:
(1) broad applicability and early diagnosis
VP1 albumen does not produce solubility capsid structure, can express the capsid of " conformation is complete ", can maintain the conformational epitope of natural viral.This conformational epitope with common capsid protein, infects extremely important for accurately detecting B19.The present invention adopts VP1 albumen as detectable antigens, can detect extensively and accurately the infection of human parvovirus B19's type.And, colloid gold label be mouse-anti people IgM monoclonal antibody, human infection human parvovirus B19 type, within about the 10th day, B19 specific IgM antibodies can be detected in infection, and continue about more than 5 months, early than the appearance of specific IgG antibodies, can early detection virus infections, be of value to early find, early treatment.
(2) detect fast and convenient: this kit detect integrated operation only need 15-20 minute, only need drip sample to sample pad, wait for result.And assist without the need to specific experimental apparatus, do not need professional training, be suitable for clinical, use at home, can Rapid Screening patient, be suitable for on-the-spot general examination, epidemiology survey etc.
(3) high sensitivity, specificity, accuracy and stability.
Visible, human parvovirus B19's type IgM antibody detection kit provided by the invention is suitable for early diagnosis, have quick, easy, do not need specific installation, accurately and sensitivity high.The whole running time only needs 15-20 minute.For medical diagnosis on disease, clinical detection all has significant application value.And be suitable for human parvovirus B19's type quick diagnosis reagent kit of epidemiology survey and rig-site utilization.
Those of ordinary skill in the field are to be understood that: the discussion of above any embodiment is only exemplary, and not intended to be implies that the scope of the present disclosure (comprising claim) is limited to these examples; Under thinking of the present invention, also can combine between technical characteristic in above embodiment or different embodiment, step can realize with random order, and there are other changes many of different aspect of the present invention as above, and they do not provide in details for the sake of simplicity.Therefore, within the spirit and principles in the present invention all, any omission made, amendment, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (10)
1. human parvovirus B19's type IgM antibody detection kit, it is characterized in that, comprise detector bar, described detector bar comprises golden bond pad and reaction film, described golden bond pad is coated with the mouse-anti people IgM monoclonal antibody of colloid gold label, described reaction film comprises detection line and nature controlling line, and described detection line is coated with restructuring HPVB19 antigen, and described nature controlling line is coated with sheep anti-mouse igg polyclonal antibody.
2. human parvovirus B19's type IgM antibody detection kit according to claim 1, it is characterized in that, described restructuring HPVB19 antigen is the VP1 albumen of human parvovirus B19's type of the recombinant expressed acquisition of using gene engineering method, and concentration is 2.0-10.0mg/mL.
3. human parvovirus B19's type IgM antibody detection kit according to claim 2, is characterized in that, the PCR primer preparing described restructuring HPVB19 antigen used is:
5'-AGCCATATGAGTAAAGAAAGTGGCAAATGGT-3'；
5'-GCACTCGAGTTATGTACCTCCTGTACCTAAAAG-3'。
4. human parvovirus B19's type IgM antibody detection kit according to claim 1, is characterized in that, described sheep anti-mouse igg Anti-TNF-α bulk concentration is 4.0-10.0mg/mL, and described mouse-anti people IgM monoclonal antibody concentration is 2.0-10.0mg/mL.
5. human parvovirus B19's type IgM antibody detection kit according to claim 1, it is characterized in that, also comprise Sample dilution, this Sample dilution is phosphate buffer.
6. human parvovirus B19's type IgM antibody detection kit according to claim 5, it is characterized in that, described Sample dilution pH value is 7.3-7.5, and conductivity is 8000 μ s/cm-12000 μ s/cm.
7. the human parvovirus B19's type IgM antibody detection kit according to claim 1 to 6 any one, it is characterized in that, described detector bar is positioned in rectangular plastic magazine, form test card, this test card is provided with sample well and detect aperture, described sample well is corresponding with the described sample pad location of described detector bar, the described detection line of the corresponding described detector bar of described detect aperture and described nature controlling line.
8. a preparation method for human parvovirus B19's type IgM antibody detection kit as claimed in claim 1, is characterized in that, comprise the following steps:
(1) preparation of reaction film:
Dilute described restructuring HPVB19 antigen and described sheep anti-mouse igg polyclonal antibody, and be sprayed on described reaction film, form described detection line and nature controlling line respectively, drying for standby;
(2) preparation of golden bond pad:
Described mouse-anti people IgM monoclonal antibody and collaurum are carried out the mouse-anti people IgM monoclonal antibody solution that coupling obtains colloid gold label, adjustment concentration, and soak described golden bond pad, paving gold, drying for standby;
(3) cutting assembling: described reaction film, golden bond pad, robust fibre filter paper, sample pad are assembled to described reaction plate, forms described detector bar after cutting.
9. the preparation method of human parvovirus B19's type IgM antibody detection kit according to claim 8, is characterized in that, the preparation method of the mouse-anti people IgM monoclonal antibody of described colloid gold label comprises the following steps:
(1) solution of potassium carbonate is used to regulate collaurum pH value to 8.0-9.0;
(2) add mouse-anti people IgM monoclonal antibody, stir;
(3) add 10% bovine serum albumin solution, stir;
(4) centrifugal, abandoning supernatant;
(5) add colloidal gold conjugate dilution, to obtain final product.
10. the preparation method of human parvovirus B19's type IgM antibody detection kit according to claim 8, it is characterized in that, the preparation method of described collaurum boils for adding gold chloride in purified water, adds citric acid three sodium solution under agitation, boils, and treats that it cools naturally.
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| CN111521786A (en) * | 2020-04-28 | 2020-08-11 | 天津健博生物科技有限公司 | Respiratory tract pathogen IgM antibody joint detection kit and preparation method thereof |
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