CN101533025A - Chemoluminescent immunoassay kit of hepatitis A virus IgM antibody and preparation method thereof - Google Patents
Chemoluminescent immunoassay kit of hepatitis A virus IgM antibody and preparation method thereof Download PDFInfo
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- CN101533025A CN101533025A CN200810101998A CN200810101998A CN101533025A CN 101533025 A CN101533025 A CN 101533025A CN 200810101998 A CN200810101998 A CN 200810101998A CN 200810101998 A CN200810101998 A CN 200810101998A CN 101533025 A CN101533025 A CN 101533025A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention relates to the field of immunoassay medical science, and in particular provides a chemoluminescent immunoassay kit of a hepatitis A virus IgM antibody and a preparation method thereof. The kit comprises: 1) negative and positive control varieties of the hepatitis A virus IgM antibody; 2) a dermatate solid phase carrier; 3) a hepatitis A virus antigen liquid; 4) an enzyme label; 5) a chemoluminescent substrate; and 6) a condensed washing solution. Furthermore, the method for preparing the kit comprises the following steps: 1) preparing control varieties from negative and positive serum of the hepatitis A virus IgM antibody; 2) dermatating the solid phase carrier by an anti-human-mu chain antibody (monoclonal antibody or polyclonal antibody); 3) preparing an antigen liquid from the hepatitis A virus; 4) labeling the hepatitis A virus antibody (the monoclonal antibody or the polyclonal antibody) by enzyme; 5) preparing the chemoluminescent substrate solution; 6) preparing the condensed washing solution; 7) sub-packaging the negative and positive control varieties of the hepatitis A virus IgM antibody, the hepatitis A virus antigen liquid, the enzyme label, the chemoluminescent substrate solution and the condensed washing solution; and 8) assembling the substances into the finished products. The kit has the advantages of simpleness, convenience, quickness, sensitiveness, stability, and the like.
Description
Technical field
The present invention relates to the immunoassay medical domain, particularly, the invention provides a kind of Chemoluminescent immunoassay kit of hepatitis A virus IgM antibody and preparation method thereof.
Background technology
Hepatitis A is to propagate a kind of communicable disease based on liver damage that causes by fecal oral route by hepatitis A virus (HAV).Be a kind of RNA viruses, belong to pico+ribonucleic acid+virus section, be enterovirus 72 types, the about 27nm of diameter spherical in shape forms 20 body nucleocapsids of symmetry by 32 shell particulates, includes the line style single-stranded RNA.HAV has 4 main polypeptide, for constituting the major antigen polypeptide of virus coat protein, induces neutralizing antibody.HAV is present in patient's blood, ight soil and the liver endochylema.Infect anti--very fast appearance of HAV1gM antibody in the serum of back, about 2 weeks, reach the peak.Descending gradually then, disappear within 8 weeks, is the serological evidence of HAV acute infection and recent infection; Have the diagnostic value higher, the early diagnosis and the prevention of hepatitis A had significance than HAVIgG antibody.
Hepatitis A virus (HAV) IgM antibody laboratory detection method is mainly immunoassay, and detection method commonly used is radiommunoassay (RIA) and EIA enzyme immunoassay (EIA).Use at present the widest be EIA enzyme immunoassay (EIA).
The research of labelled immune analytical technology and application development are rapid in recent years, have been widely used in each field of biomedical fundamental research and clinical disease diagnosis.What be easy to promote mainly contains: four kinds of radiommunoassay, EIA enzyme immunoassay, time resolved fluoro-immunoassay and chemiluminescence immune assays etc.The basic theories of these ultramicron detection techniques is identical substantially, but used tracer agent and the signal that sent have nothing in common with each other.According to a large amount of test findings and clinical practice data,, be followed successively by: chemiluminescence immune assay, time resolved fluoro-immunoassay, radiommunoassay and EIA enzyme immunoassay from practicality, stability, accuracy and development prospect.Chemiluminescence immunoassay technology, the high sensitivity that had both had methods such as radiommunoassay, time resolved fluoro-immunoassay, the advantage that the range of linearity is wide, shortcomings such as the short and time resolved fluoro-immunoassay poor stability of the radioactive contamination, half life period of radiommunoassay, instrument costliness have been overcome again, can be widely used in biomedical research and clinical medical inspection, be more convenient for applying in large, medium and small hospital.
At present, chemiluminescence immunoassay technology is just extensively being used aspect the in-vitro diagnosis immunological assay reagents, but adopts direct coated antigen not see use in conjunction with chemiluminescence immunoassay technology yet in the application facet of antihepatitis A virus IgM immunoassay product.Kit of the present invention adopts the microwell plate chemiluminescence immunoassay technology, both can be applicable to the open luminous measuring instrument of semi-automatic chemistry, also can be used for full automatic measuring system, can realize fast detection the in enormous quantities, and use cost is low, easier applying.
Summary of the invention
The present invention has solved the problems referred to above simultaneously well, chemiluminescence learned effectively with the immunoassay of antihepatitis A virus IgM combine, a kind of kit that can easy, quick, sensitive, stably detect antihepatitis A virus IgM is provided, and this kit is suitable for promotion and application effectively on industry.
The purpose of this invention is to provide a kind of chemiluminescence enzyme immunoassay and measure kit of antihepatitis A virus IgM and preparation method thereof.
Kit according to the present invention comprises: 1) antihepatitis A virus IgM positive and negative reference substance; 2) solid phase carrier of bag quilt; 3) hepatitis A viral antigen liquid; 4) anti-HAV of enzyme labeling (monoclonal antibody or how anti-); 5) chemical luminous substrate that above-mentioned enzyme acted on; And 6) concentrated cleaning solution.
According to kit of the present invention, wherein, described solid phase carrier is microwell plate, plastic bead, plastic tube or magnetic-particle; Described marker enzyme is alkaline phosphatase or horseradish peroxidase; Described chemical luminous substrate is 1,2-two oxidative ethane analog derivatives, luminol or different luminol, wherein said 1,2-two oxidative ethane analog derivatives are (diamantane)-1,2-two oxidative ethanes, 3-(2 '-the spiral diamantane)-4-methoxyl-4-(3 " the phosphorus acyloxy) phenyl-1,2-two oxidative ethanes (AMPPD), CSPD or CDP-Star.
Further, the invention provides a kind of method for preparing the mentioned reagent box, may further comprise the steps:
1) with antihepatitis A virus IgM positive and negative serum preparation reference substance;
2) wrap by solid phase carrier with anti-people-μ chain antibody (monoclonal antibody or how anti-);
3) prepare antigen liquid with hepatitis A viral antigen;
4) with enzyme labeling anti-HAV (monoclonal antibody or how anti-);
5) preparation chemical luminous substrate liquid;
6) preparation concentrated cleaning solution
7) chemical luminous substrate that anti-HAV, this enzyme acted on and the concentrated cleaning solution of the above-mentioned antihepatitis A virus IgM positive and negative of packing reference substance, hepatitis A viral antigen liquid, enzyme labeling; And
8) be assembled into finished product.
The method according to this invention, preferred, described bag is by the step 2 of solid phase carrier) may further comprise the steps:
1) bag quilt
With 0.1M pH value is anti-people-μ chain antibody (monoclonal antibody or how anti-) coating buffer that 9.0 dipotassium hydrogen phosphate damping fluid is mixed with desired concn, and coating buffer is carried on the solid phase carrier;
2) sealing
With containing 2% BSA, 0.01% gelatin, the pH value is 6.8～7.3, concentration is that the phosphate buffer of 0.01M is as the above-mentioned solid phase carrier of confining liquid sealing.
In said method, preferred, the solid phase carrier of described antibody sandwich is microwell plate, plastic bead, plastic tube or magnetic-particle; Described bond is horseradish peroxidase, alkaline phosphatase; Described chemical luminous substrate is 1,2-two oxidative ethane analog derivatives, luminol or different luminol, wherein said 1,2-two oxidative ethane analog derivatives are (diamantane)-1,2-two oxidative ethanes, 3-(2 '-the spiral diamantane)-4-methoxyl-4-(3 " the phosphorus acyloxy) phenyl-1,2-two oxidative ethanes, CSPD or CDP-Star.
Concrete mentioned reagent box can comprise microwell plate, hepatitis A viral antigen liquid, enzyme labeling thing and chemical luminous substrate liquid, concentrated cleaning solution of antihepatitis A virus IgM positive and negative reference substance, the anti-people of direct coated-μ chain antibody etc.Wherein, described antihepatitis A virus IgM positive and negative reference substance is through HBsAg, anti-HIV, anti--TP and many parts of negative pooled serums of anti-HCV TPPA, 60 ℃ of deactivations 1 hour, (anti--HAVIgM antibody positive is tired〉1:1000) appropriateness dilution system is joined.The microwell plate of the anti-people of direct coated-μ chain antibody is the micropore lath in 48 or 96 holes, the enzyme labeling thing for coupling horseradish peroxidase (HRP), chemical luminous substrate liquid is luminol, concentrated cleaning solution is PBST.
The present invention's " antihepatitis A virus IgM chemical luminescence immune assay determination reagent kit " can detect the antihepatitis A virus IgM in the human body behind the early infection hepatitis A virus very single-mindedly, can judge the variation of the result of treatment and the early stage state of an illness thereof according to the antihepatitis A virus IgM situation of change.It has advantages such as easy, quick, sensitive, stable.And detection system of the present invention is an open-sky technique, does not easyly fast need the expensive luminous measuring instrument of full-automatic chemical, is particularly suitable for vast middle and small hospital and promotes the use of, for clinical diagnosis and research work provide a kind of very valuable detection means.According to kit of the present invention, anti-people-μ the chain antibody of bag quilt is caught specific antibody in the sample (IgM antibody) earlier on the carrier, again in conjunction with the hepatitis A virus specific antigen that adds, the specific antibody of last and enzyme labeling is in conjunction with the formation antigen antibody complex, so " prize law " reaction pattern of the present invention's employing, both effectively utilize the chemiluminescence principle, guaranteed the sensitivity that detects again.In addition, kit of the present invention is convenient to large-scale production, is convenient to application in the reality.
What kit of the present invention was used is the enzymatic luminous substrate, by the chromogenic substrate in the light signal replacement EIA enzyme immunoassay that detects the luminous substrate generation, thereby have a specificity equal with EIA enzyme immunoassay, and it is highly sensitive, detect the serum sample ash area value than ELISA reagent and lack, avoided detecting sample and judged the probability that false positive and negative result's appearance occurs.The diagnosis that can be antihepatitis A virus IgM provides more special, quick, reliable foundation.
Embodiment
Embodiment 1 preparation antihepatitis A virus IgM chemical luminescence immune assay determination reagent kit of the present invention
In research process of the present invention, the present inventor has at first carried out screening experiment and Quality Identification to used starting material, comprises the luminous intensity of activity, chemical luminous substrate of the absorption property of activity, carrier (as lighttight white microwell plate) of antibody labeling thing and coated antibody and variation size, HRP and luminous duration etc.Then method for coating is studied, be cushioned liquid and protect liquid to experimentize, select optimal bag and be cushioned liquid and protection liquid, tested by concentration by the different bags of antibody and find best concentration conditions with different bags.Mark for HRP can have diverse ways, by explore repeatedly and the contrast experiment finally found easy, productive rate is high, cost is low, the reliable quality labeling method, and different enzyme dilutions has been carried out long-term investigation tested, made and can make the activity stabilized dilution of the long-term maintenance of antibody labeling thing.
One, enzyme labelled antibody preparation
The antibody of hepatitis A viral antigen is fully dialysed with PBS with glutaraldehyde method and horseradish peroxidase, adds equal-volume glycerine, preserves below-20 ℃.
Two, the preparation of antihepatitis A virus IgM positive and negative contrast
Mix more than 6 parts and detect antihepatitis A virus IgM positive serum or negative serum through the ELISA kit, through HBsAg, anti-HIV, anti--TP and anti-HCV negative antibody serum, 60 ℃ of deactivations 1 hour, filtration sterilization, appropriateness dilution, packing, low temperature is frozen.
Three, enzyme labelled antibody concentration is selected
The configuration of enzyme mark monoclonal antibody dilution:
Borax 1.4304g
Boric acid 5.2564g
Calf serum 200ml
Glycerine 20ml
Proclin300 1ml
Distilled water 1000ml
With enzymic-labelled antibody with the enzyme diluted different working concentration, utilize the square formation titrimetry to select the working concentration that enzyme labelled antibody is fit to and be 1:5500.
Four, the preparation of solid-phase coating plate
(1) bag quilt
K
2HPO
4·3H
2O 22.823g
Distilled water 1000ml
Behind the dissolving mixing, adjust PH to 9.0, add an amount of anti-people-μ chain antibody mixing, add then in each hole of microwell plate, every hole 100 μ L, 4 ℃ are spent the night.
(2) sealing
NaH
2PO
4·2H
2O 0.6944g
Na
2HPO
4·12H
2O 2.1851g
NaCL 8.5g
BSA 20g
Gelatin 0.1g
Proclin 3001ml
Distilled water is settled to 1000ml
The mentioned reagent weighing is put into clean container well, add the distilled water constant volume, the dissolving mixing, measuring the pH value is 6.8～7.3.
Every hole adds confining liquid 150 μ L respectively, adsorbs 24 ℃ hours.Get rid of confining liquid, on thieving paper, pat dry.Room temperature removal moisture drying 24 hours.Carry out vacuum sealing bag immediately.Place behind the envelope and checked not have gas leakage in 15 minutes, if there is gas leakage to need envelope again.2～8 ℃ of preservations of labeling postposition.
Five, chemical luminous substrate A liquid
Luminol 10mM 1.7716g
4-xenol 0.3mM 0.051g
4-iodobenzene boric acid 0.05mM 0.012g
Boric acid 11.4g
Borax 4.9g
The fixed molten 1000mL of distilled water
pH?8.0～10.0
Six, chemical luminous substrate B liquid
Urea peroxide 3.5mM 0.329g
Tween20 0.1％ 1ml
Na
2HPO
4·12H
2O 51.58g
NaH
2PO
4·2H
2O 8.74g
The fixed molten 1000mL of distilled water
pH?7.0～7.6
A liquid mixes with B liquid equal proportion during use.
Seven, 20 times of cleansing solutions
Na
2HPO
4·12H
2O 58g
NaH
2PO
4·2H
2O 2g
NaCl 160g
Tween-20 1mL
Proclin?300 1mL
Deionized water 1000mL
Adjust pH to 7.2～7.4
Eight, semi-manufacture and finished product are formed
The packing of above-mentioned steps products obtained therefrom is semi-manufacture.Extract three parts of process specificitys, accuracy, sensitivity and stable assay approvals out and just can be assembled into antihepatitis A virus IgM mensuration kit (chemoluminescence method).Be assembled into also need inspect by random samples behind the kit and just can dispatch from the factory after qualified.
The present inventor has also carried out experimental study to the reaction pattern and the reaction conditions of kit, has finally determined the prize law reaction pattern, and the influence of experimental result is tested with regard to the different reaction time, determines the optimal reaction time.By the influence experiment to measured value shows to the mensuration of luminous duration of chemical luminous substrate liquid and different fluorescent lifetime: be measured as the best between 5-25 minute after adding chemical luminous substrate liquid, its result is also comparatively accurate.
Embodiment 2～3 preparations antihepatitis A virus IgM chemical luminescence immune assay determination reagent kit of the present invention
Divided by the alkali phosphatase enzyme mark anti-HAV, with AMPPD as chemical luminous substrate, with plastic bead, plastic tube as the pH value of carrier, coating buffer be 4.5, the pH value of confining liquid is outside 7.5, all the other all prepare the antihepatitis A virus IgM chemical luminescence immune assay determination reagent kit with the method identical with embodiment 1.
Embodiment 4 preparations antihepatitis A virus IgM chemical luminescence immune assay determination reagent kit of the present invention
, prepare outside the anti-people of magnetic particle-μ chain antibody solid phase carrier with the glutaraldehyde coupling method as solid phase carrier divided by magnetic-particle, all the other all prepare the antihepatitis A virus IgM chemical luminescence immune assay determination reagent kit with the method identical with embodiment 1.
The using method of embodiment 5 kits of the present invention
The concrete operations of the antihepatitis A virus IgM chemical luminescence immune assay determination reagent kit of above embodiment 1 preparation are as follows:
1) in 4 ℃ of refrigerators, takes out kit, equilibrium at room temperature 15 minutes.To concentrate again thin liquid dilute in proportion for working concentration standby.
2) earlier blood serum sample to be checked is used physiological saline 1:1000 diluted for use
3) take out coated slab, insert on the grillage.
4) except that blank well, add each two hole of yin, yang reference substance in the reacting hole respectively, every hole 50ul, each hole adds sample 50ul to be checked, and every then hole adds antigen 50 μ L, with the micro-oscillator mixing that fully vibrates, 37 ℃ of incubations 0.5 hour.
5) get rid of dereaction liquid, the cleansing solution after the dilution is filled it up with in every hole, washes plate 5 times, buckles to do on clean thieving paper, adds enzyme labelled antibody label 100 μ L then, with the micro-oscillator mixing that fully vibrates, and 37 ℃ of incubations 0.5 hour.
6) get rid of dereaction liquid, the cleansing solution after the dilution is filled it up with in every hole, washes plate 5 times, buckles on clean thieving paper at last and does.
7) each hole adds each 50 μ L of chemical luminous substrate liquid A, B, and fully vibrating with micro-oscillator mixes, room temperature (20～25 ℃) lucifuge reaction 5 minutes.
8) must measure in the 5th～25 minute after adding chemical luminous substrate liquid, on the chemiluminescence measuring instrument, measure the luminous intensity (RLU) in each hole in regular turn, 1 second/hole of Measuring Time.
9) CO value=2.1N, RLU value 〉=CO value then sample is judged to be positive sample; RLU value＜CO value then sample is judged to be negative sample.
The methodology of embodiment 6 kits of the present invention is identified
According to manufacturing conventional in this area and vertification regulation the kit of preparation among the embodiment 1 is examined and determine, is seen the following form 1:
Table 1
Sensitivity, specificity, accuracy and stability that the present invention's " antihepatitis A virus IgM (anti--HAVIgM) chemical luminescence method immune analysis is measured kit " is described meet the calibrating requirement fully.
Embodiment 7 antihepatitis A virus IgMs of the present invention (anti--HAVIgM) clinical practice and the ELISA kit of chemical luminescence method immune analysis mensuration kit compare
Two tame clinical hospitals use embodiment 1 antihepatitis A virus IgM (anti--HAVIgM) chemical luminescence method immune analysis measure kit and ELISA kit (this kit for obtain State Food and Drug Administration produce the detection of official written reply anti--the ELISA product of HAVIgM) detect 712 parts of clinical serum sample (310 parts of healthy people, healthy children 60 examples, HBsAg positive patient 120 examples, erythematosus lupus patient 40 examples, positive 65 examples of rheumatoid factor, cord serum 35 examples) and 102 routine clinical definites be hepatitis A patient patients serum sample, to contrast the coincidence rate that two kinds of kits detect.
One, the source of clinical serum specimen
Hepatitis A virus that each hospital clinical is collected and non-hepatitis A virus patient and normal person's sample.
Two, use antihepatitis A virus IgM (anti--HAVIgM) chemical luminescence method immune analysis mensuration kit and the comparison of ELISA kit
1, method
Two tame hospitals use the embodiment of the invention 1 preparation " antihepatitis A virus IgM (anti--HAVIgM) chemical luminescence method immune analysis is measured kit " (by its instructions operation) and ELISA kit (by its instructions operation) to detect 712 parts of clinical samples respectively, to contrast the coincidence rate of two kinds of kits detections.
2, result
The two tame use embodiment 1 of hospitals " antihepatitis A virus IgM (anti--HAVIgM) chemical luminescence method immune analysis is measured kit " and ELISA kit comparison and detection result (seeing Table 2)
Table 2. liang two kinds of methods of tame hospital's contrast relatively
The use of two tame hospitals " antihepatitis A virus IgM (anti--HAVIgM) chemical luminescence method immune analysis is measured kit " and show with ELISA kit comparison and detection result: the specificity 100% of kit of the present invention, susceptibility 100%; In the clinical testing, contrast agent 2 routine rheumatoid factor positive sample serum detect the HAVIgM antibody positive and 1 routine hepatitis A positive sample detects the HAVIgM negative antibody, 3 parts of serum sample repetition measurement validation tests after 2 mercapto ethanol is handled, 2 routine rheumatoid factor positive sample results are negative, and 1 routine hepatitis A positive sample repetition measurement result is positive.This kit and rheumatoid factor positive serum, hepatitis B virus surface antigen positive serum no cross reaction.The antihepatitis A virus IgM of invention (anti--HAVIgM) chemical luminescence immune assay determination reagent kit specificity, sensitivity, be better than contrast agent, can be applied to the diagnostic detection of clinical hepatitis A early infection and acute hepatitis A index.
Claims (10)
1, a kind of antihepatitis A virus IgM chemical luminescence immune assay determination reagent kit is characterized in that, described kit comprises:
1) antihepatitis A virus IgM positive and negative reference substance;
2) solid phase carrier of bag quilt;
3) hepatitis A viral antigen liquid;
4) enzyme labeling thing;
5) chemical luminous substrate that above-mentioned enzyme acted on; And
6) concentrated cleaning solution.
2, kit as claimed in claim 1 is characterized in that, described solid phase carrier is microwell plate, plastic bead, plastic tube or the magnetic-particle of anti-people-μ chain antibody (monoclonal antibody or how anti-) direct coated.
3, kit as claimed in claim 1 is characterized in that, described enzyme labeling thing is the anti-HAV (monoclonal antibody or how anti-) with alkaline phosphatase or horseradish peroxidase-labeled.
4, kit as claimed in claim 1 is characterized in that, described chemical luminous substrate is 1,2-two oxidative ethane analog derivatives, luminol or different luminol.
5, kit as claimed in claim 1 is characterized in that, described chemical luminous substrate comprises A liquid and B liquid, wherein,
Described chemical luminous substrate A liquid comprises 10mM luminol, 0.3mM 4-xenol, 0.05mM 4-iodine substituted phenyl boric acid, 0.2M pH8.7 boric acid-borate buffer solution, and its pH value is 8.0～10.0;
Described chemical luminous substrate B liquid comprises 3.5mM urea peroxide, 0.1%Tween20,0.2M pH7.2 phosphate buffer, and its pH value is 7.0～7.6.
6, a kind of method for preparing the described kit of claim 1 is characterized in that may further comprise the steps:
1) with antihepatitis A virus IgM positive and negative serum preparation reference substance;
2) with anti-people-μ chain antibody (monoclonal antibody or how anti-) direct coated solid phase carrier;
3) prepare antigen liquid with hepatitis A viral antigen;
4) with enzyme labeling anti-HAV (monoclonal antibody or how anti-);
5) preparation chemical luminous substrate liquid;
6) preparation concentrated cleaning solution;
7) chemical luminous substrate that anti-HAV, this enzyme acted on and the concentrated cleaning solution of the above-mentioned antihepatitis A virus IgM positive and negative of packing reference substance, hepatitis A viral antigen liquid, enzyme labeling; And
8) be assembled into finished product.
7, method as claimed in claim 6 is characterized in that, described bag is by the step 2 of solid phase carrier) adopt following method:
1) bag quilt
With 0.1M pH value is anti-people-μ chain antibody (monoclonal antibody or how anti-) coating buffer that 9.0 dipotassium hydrogen phosphate damping fluid is mixed with desired concn, and coating buffer is carried on the solid phase carrier;
2) sealing
With containing 2% BSA, 0.01% gelatin pH value is 6.8～7.3, and concentration is that the phosphate buffer of 0.01M seals above-mentioned solid phase carrier as confining liquid.
As claim 6 or 7 described methods, it is characterized in that 8, described bag is microwell plate, plastic bead, plastic tube or magnetic-particle by anti-people-μ chain antibody solid phase carrier.
9, method as claimed in claim 6 is characterized in that, described enzyme is alkaline phosphatase or horseradish peroxidase; Described chemical luminous substrate is 1,2-two oxidative ethane analog derivatives, luminol or different luminol.
10, method as claimed in claim 9, it is characterized in that described 1,2-two oxidative ethane analog derivatives are (diamantane)-1,2-two oxidative ethanes, 3-(2 '-the spiral diamantane)-4-methoxyl-4-(3 " the phosphorus acyloxy) phenyl-1,2-two oxidative ethanes, CSPD or CDP-Star.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810101998A CN101533025A (en) | 2008-03-14 | 2008-03-14 | Chemoluminescent immunoassay kit of hepatitis A virus IgM antibody and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810101998A CN101533025A (en) | 2008-03-14 | 2008-03-14 | Chemoluminescent immunoassay kit of hepatitis A virus IgM antibody and preparation method thereof |
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| CN101533025A true CN101533025A (en) | 2009-09-16 |
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| CN200810101998A Pending CN101533025A (en) | 2008-03-14 | 2008-03-14 | Chemoluminescent immunoassay kit of hepatitis A virus IgM antibody and preparation method thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102353777A (en) * | 2011-07-05 | 2012-02-15 | 深圳市卫武光明生物制品有限公司 | Kit and method for testing human cytomegalovirus IgG antibody |
| CN102692502A (en) * | 2012-04-26 | 2012-09-26 | 北京北方生物技术研究所 | Rapid hepatitis A virus IgM antibody detection kit and preparation method thereof |
| CN108169475A (en) * | 2017-12-18 | 2018-06-15 | 郑州安图生物工程股份有限公司 | A kind of Respiratory Syncytial Virus(RSV) IgM antibody detection kit |
| CN109946464A (en) * | 2019-04-26 | 2019-06-28 | 江苏硕世生物科技股份有限公司 | Detect ELISA Plate, kit and the preparation method of antihepatitis A virus IgM |
| CN110672850A (en) * | 2019-10-31 | 2020-01-10 | 迪瑞医疗科技股份有限公司 | Hepatitis A virus antibody IgM detection kit and preparation method thereof |
-
2008
- 2008-03-14 CN CN200810101998A patent/CN101533025A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102353777A (en) * | 2011-07-05 | 2012-02-15 | 深圳市卫武光明生物制品有限公司 | Kit and method for testing human cytomegalovirus IgG antibody |
| CN102353777B (en) * | 2011-07-05 | 2014-07-02 | 深圳市卫光生物制品股份有限公司 | Kit and method for testing human cytomegalovirus IgG antibody |
| CN102692502A (en) * | 2012-04-26 | 2012-09-26 | 北京北方生物技术研究所 | Rapid hepatitis A virus IgM antibody detection kit and preparation method thereof |
| CN102692502B (en) * | 2012-04-26 | 2015-02-04 | 北京北方生物技术研究所 | Rapid hepatitis A virus IgM antibody detection kit and preparation method thereof |
| CN108169475A (en) * | 2017-12-18 | 2018-06-15 | 郑州安图生物工程股份有限公司 | A kind of Respiratory Syncytial Virus(RSV) IgM antibody detection kit |
| CN109946464A (en) * | 2019-04-26 | 2019-06-28 | 江苏硕世生物科技股份有限公司 | Detect ELISA Plate, kit and the preparation method of antihepatitis A virus IgM |
| CN110672850A (en) * | 2019-10-31 | 2020-01-10 | 迪瑞医疗科技股份有限公司 | Hepatitis A virus antibody IgM detection kit and preparation method thereof |
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