CN101551395A - Chemiluminescence immunoassay kit of hepatitis E virus IgM antibody and preparation method thereof - Google Patents
Chemiluminescence immunoassay kit of hepatitis E virus IgM antibody and preparation method thereof Download PDFInfo
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- CN101551395A CN101551395A CNA2008101032644A CN200810103264A CN101551395A CN 101551395 A CN101551395 A CN 101551395A CN A2008101032644 A CNA2008101032644 A CN A2008101032644A CN 200810103264 A CN200810103264 A CN 200810103264A CN 101551395 A CN101551395 A CN 101551395A
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Abstract
The invention relates to a chemiluminescence immunoassay kit of a hepatitis E virus IgM antibody and a preparation method thereof, which belongs to the technical field of clinical in-vitro diagnosis immunoassay. The preparation method of the kit comprises the following steps: the reference substance is prepared according to the negative and the positive serums of the hepatitis E virus IgM antibody; a solid phase carrier is coated by an anti-human-mu chain anti-body (monoclonal antibody or polyclonal antibody); the recombinant antigen of the hepatitis E virus is labeled by enzyme; a chemiluminescent substrate solution acted by the enzyme is prepared; a concentrated cleaning solution is prepared; the negative reference substance, the positive reference substance, the labeled combination, the chemiluminescent substrate and the concentrated cleaning solution of the hepatitis E virus IgM antibody are subpackaged; and all the components are assembled into finished products. The prepared kit has stable performance indexes (specificity, sensitivity and stability) and can be applied to the early diagnosis of the hepatitis E.
Description
Technical field
The present invention relates to clinical in vitro diagnosis in vitro immunoassay medical domain, particularly, the invention provides a kind of chemiluminescence immunoassay kit of hepatitis E virus IgM antibody and preparation method thereof.
Background technology
(hepatitis E virus is nonencapsulated single positive chain RNA virus HEV) to hepatitis E virus, propagates through fecal-oral route, acute Hepatitis E (the hepatitisE that causes, HE), show as acute Jaundice Jaundice more, easily develop into subacute hepatitis sequestrans or silt courage hepatitis.Be ascendant trend year by year in recent years.The patient is mainly the adult, and case fatality rate is higher, especially last 3 months gravid woman of pregnancy period ill after, case fatality rate can reach 10%～39%, harm is serious.Viral hepatitis type E at first finds that at the Indian subcontinent the Central Asia, Southeast Asia, Africa, the Indian subcontinent all have more pandemic report.Most eruption and prevalences are aquagenic.The report of food source property also is seen in document.The infection rate of population of China Hepatitis E about 18%.Viral hepatitis type E accounts for 10% in the acute sporadic hepatitis, is one of China's Class B Notifiable disease.Can measure the anti-HEV-IgM of high titre in acute phase serum, convalescence, anti-HEV-IgM titre descended or disappearance.But the substitute is and produce anti-HEV-IgG in the serum.Anti--HEV1gM occurs in the serum, is the serological evidence of HEV acute infection and recent infection; Have the diagnostic value higher than HEVIgG antibody, therefore early diagnosis and the prevention to Hepatitis E has significance.
Domestic Hepatitis E diagnostic method commonly used has at present: immunofluorescence technique, immunoelectron microscope, RT-polymerase chain reaction method (RT-PCR), specific antibody detection method.Immunofluorescence technique detects hepatitis E virus antigen in the hepatic tissue, and the method must be carried out liver and be worn biopsy; Immunoelectron microscope detects in the ight soil of patients during acute stage and the bile viral antigen or with corresponding antibody among the known patients serum; RT-polymerase chain reaction method (RT-PCR) detects hepatitis E virus RNA (ribonucleic acid) (HEV-RNA).Hepatitis E virus specific antibody euzymelinked immunosorbent assay (ELISA) detection method is the main detection method of present hospital, and it is measured anti-HEV-IgM and has clinical meaning.Because the sensitivity of ELISA reagent method not as the chemical illuminating reagent height, is infected the window phase patient to HEV and can not be diagnosed ahead of time; RT-PCR, immunofluorescence technique method, immunoelectron microscope detection method, complex operation, has limitation at poor stability, instrument costliness.Given this, the present invention set up a kind of fast, sensitivity, specificity, stable strong chemiluminescence enzyme promoting immunity analytical approach detect hepatitis E virus antibody.
Now, chemical luminescence immune analysis reagent box mostly is the luminous measuring system of the closed full-automatic chemical of import, apparatus expensive, and the use cost height is promoted the use of thereby limited, and can't be widely used in clinical diagnosis and research work.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, use cost is low, more can be adapted at large, medium and small hospital and apply.
Summary of the invention
The objective of the invention is: the kit that a kind of chemiluminscence immunoassay hepatitis E virus IgG antibody is provided.This invention kit can be easy, quick, sensitive, equipment is simple, repeatability is better, pollution-free can stably detect hepatitis E virus IgM antibody, and is suitable for applying effectively on industry.
Kit according to the present invention comprises: hepatitis E virus IgM antibody positive and negative reference substance; Bag is by solid phase carrier; The enzyme labeling bond; The chemical luminous substrate liquid that enzyme acted on; And concentrated cleaning solution.
According to kit of the present invention, wherein, described bag is microwell plate, plastic bead, plastic tube or magnetic-particle by solid phase carrier; Described enzyme is horseradish peroxidase or alkaline phosphatase; Described chemical luminous substrate liquid 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.
The present invention adopts and catches the sandwich method principle, utilizes chemiluminescence immunoassay to measure the variation of Hepatitis E IgM antibody horizontal.Bag by the anti-people of high-purity-μ chain bag by microwell plate, enzyme labeling specificity recombined hepatitis E hepatitis virus antigen.The sample to be tested, the enzyme labeling thing that add dilution in wrapping by the plate micropore form the compound of antibody-antibody-enzyme-labelled antigen admittedly behind the incubation, abundant washing back adds chemical luminous substrate liquid, measures its luminous intensity (RLU).Increase with anti--IgM antibody concentration raises according to the RLU value of sample, thereby
JudgeThe situation of change of hepatitis E virus IgM antibody in human body.
Another order of the present invention provides a kind of method for preparing the mentioned reagent box, may further comprise the steps:
1) join reference substance with hepatitis E virus IgM antibody positive and negative serum system: the hepatitis E virus IgM antibody positive and negative are through HBsAg, anti-HIV, anti--TP and many parts of all negative pooled serums of anti-HCV TPPA to serum, 60 ℃ of deactivations 1 hour, appropriateness dilution preparation (anti--HEVIgM antibody positive tire＞1: 500)
2) with anti-people-μ chain antibody (monoclonal antibody or many anti-) direct coated solid phase carrier (microwell plate, plastic bead, plastic tube or magnetic-particle): with 0.1M pH value is the hepatitis E virus antigen coating buffer that 8.0 borate buffer is mixed with desired concn, and coating buffer is carried on the solid phase carrier; With containing 0.1% casein, 1% sucrose, 0.01% gelatin, 0.1%Proclin300, the pH value is 7.0～7.5, concentration is that the phosphate buffer of 0.01M is as the above-mentioned solid phase carrier of confining liquid sealing
3) with enzyme labeling recombined hepatitis E hepatitis virus antigen: with horseradish peroxidase (HRP) or alkaline phosphatase coupling recombined hepatitis E hepatitis virus antigen;
4) the preparation chemical luminous substrate liquid that enzyme acted on; With 1,2-two oxidative ethane analog derivatives, luminol or the different luminol preparation chemical luminous substrate liquid that enzyme acted on.
5) preparation concentrated cleaning solution: for PBST concentrates thin liquid.
Chemical luminous substrate and concentrated cleaning solution that hepatitis E virus recombinant antigen, this enzyme of the above-mentioned hepatitis E virus IgM antibody positive and negative of packing reference substance, enzyme labeling acted on; And be assembled into finished product.
The present invention's " hepatitis E virus IgM antibody chemical luminescence immune assay determination reagent kit " can detect the hepatitis E virus IgM antibody in the human body after the early infection hepatitis E virus very single-mindedly, can change according to detecting hepatitis E virus IgM antibody, judge the variation of the result of treatment and the early stage state of an illness
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, recombined hepatitis E hepatitis virus specific antigen combination with the enzyme labeling that adds, form antigen antibody complex, so " catching sandwich method " reaction pattern of the present invention's employing, both effectively utilize the chemiluminescence principle, guaranteed the sensitivity that detects again.In addition, this reaction pattern also is convenient to operation.
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 has better than EIA enzyme immunoassay specificity, highly sensitive characteristics; The diagnosis that can be hepatitis E virus IgM antibody provides more special, quick, reliable foundation.
Embodiment
Embodiment 1 preparation hepatitis E virus IgM antibody 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 antigenic label 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 antigenic label.
One, horseradish peroxidase recombined hepatitis E hepatitis virus antigen preparation:
Recombined hepatitis E hepatitis virus antigen dimaleimide (Dimaleimide) coupling horseradish peroxidase.Concrete operations are as follows: will resist lgG antibody to be dissolved in 0.1Mol/L pH5.0 sodium-acetate buffer and same damping fluid dialysis equilibrium will be spent the night, and the centrifugal insoluble substance of removing, anti-IgG (14mg/2ml) and 10ml alpha-mercapto ethamine are at 37 ℃ of incubation 90min.Crossing to concentrate behind the Sephadex makes every ml contain the anti-IgG of reduction about 3.0mg.In 0 ℃, dripped saturated N, N '-O-phenylenedimaleimide solution by 1: 1, mix, in 30 ℃ of incubation 20min, cross SephadexG
25Post is removed unreacted dimaleimide.Collect the anti-IgG of dimaleimide " activation ", concentrating and making concentration is OD
280nm=1.0 (optical path 1cm) get 1ml solution and add 20 μ l (5mg/ml) horseradish peroxidases, put 30 ℃ of 20min, with among the 0.10Mol/L NaOH and after, put 24h～72h in 4 ℃, after Sepharose-6B post (1.5 * 40cm), with the pH7.0PBS wash-out, collect enzyme conjugates, add Sodium azide (NaN
3) anticorrosion, 4 ℃ of preservations are standby.
Enzyme labeling recombined hepatitis E hepatitis virus antigen concentration is selected
Based on 1000ml enzyme-labelled antigen dilution component content be: 2.9g Na
2HPO
412H
2O, 0.3gNaH
2PO
42H
2O, 9g NaCl, 25g BSA, 1ml Proclin300,1ml Tween-20.
With the different dilutability of enzyme-labelled antigen bond dilution, adopt the square formation titrimetry to determine that the working concentration of enzyme-labelled antigen is 1: 6000
Two, the preparation of hepatitis E virus IgM antibody positive and negative contrast
Mix more than 6 parts and detect hepatitis E virus IgM antibody positive serum or negative serum through the ELISA kit, through anti-HIV, anti--TP and anti-HCV negative antibody serum, 60 ℃ of deactivations 1 hour, appropriateness dilution preparation (anti--HEVIgM antibody positive tire＞1: 500), filtration sterilization, packing, low temperature is frozen.
Three, the preparation of solid-phase coating plate
The bag quilt: based on 1000ml coating buffer component content is 2.86g borax, 4.33g boric acid, takes by weighing after required component amount puts into clean container distilled water dissolving mixing, is adjusted to PH 8.0, fixed molten 1000ml.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.Bag is the micropore lath in 48 or 96 holes by microwell plate.
Sealing: every hole adds confining liquid 150 μ L respectively, places 24 hours for 4 ℃.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.
Based on 1000ml confining liquid component content be: 0.2g NaH
2PO
42H
2O, 2.9g Na
2HPO
412H
2O, 1g casein, 10g sucrose, 0.1g gelatin, 1ml Proclin300.The good each component amount of weighing is put into clean container, adds distilled water, the dissolving mixing, and adjusting the pH value is 7.0～7.5, constant volume 1000ml.
Four, chemical luminous substrate A liquid
Based on 1000mlA liquid component content be: 10mM luminol 1.7716g, 0.3mM 4-xenol 0.051g, 0.05mM 4-iodobenzene boric acid 0.012g, boric acid 11.4g, borax 4.9g.The good each component amount of weighing is put into clean container, adds distilled water, the dissolving mixing, and adjusting the pH value is 8.0～10.0, constant volume 1000ml.
Five, chemical luminous substrate B liquid
Based on 1000mlA liquid component content be: 3.5mM urea peroxide 0.329g, 1ml 0.1%Tween20,51.58gNa
2HPO
412H
2O, 8.74g NaH
2PO
42H
2O, the good each component amount of weighing are put into clean container, add distilled water, the dissolving mixing, and adjusting the pH value is 7.0～7.6, constant volume 1000ml.
The good each component amount of weighing is put into clean container, adds distilled water, the dissolving mixing, and adjusting the pH value is 8.0～10.0, constant volume 1000ml.
A liquid mixes with B liquid equal proportion during use.
Six, 20 times of cleansing solutions
Based on 20 times of cleansing solution each components of 1000ml content is 58g Na
2HPO
412H
2O, 2g NaH
2PO
42H
2O, 160g NaCl, 1mL Tween-20,1mL Proclin 300.The good each component amount of weighing is put into clean container, adds distilled water, the dissolving mixing, and adjusting the pH value is 7.2～7.4, constant volume 1000ml.
Seven, semi-manufacture and finished product are formed
The packing of above-mentioned steps products obtained therefrom is semi-manufacture.Semi-manufacture just can be assembled into hepatitis E virus IgM antibody through specificity, accuracy, sensitivity and stable assay approval and measure kit (chemoluminescence method) finished product.Being assembled into the finished product kit can use behind specificity, accuracy, sensitivity and stable assay approval again.
The present inventor has finally determined the sandwich reaction pattern of prize law by the experimental study that reaction pattern and reaction conditions to kit carry out, 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 hepatitis E virus IgM antibody chemical luminescence immune assay determination reagent kit of the present invention
Divided by alkali phosphatase enzyme mark hepatitis E virus recombinant antigen, 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 hepatitis E virus IgM antibody chemical luminescence immune assay determination reagent kit with the method identical with embodiment 1.
Embodiment 4 preparations hepatitis E virus IgM antibody 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 carrier divided by magnetic-particle, all the other all prepare the hepatitis E virus IgM antibody 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 hepatitis E virus IgM antibody 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.Serum sample 1: 1000 diluted for use of physiological saline.
2) take out coated slab, insert on the grillage.
3) except that blank well, add each two hole of yin, yang reference substance in the reacting hole respectively, every hole 50ul, each hole adds the sample 50ul to be checked after the dilution, and every then hole adds enzyme conjugates 50 μ L, with the micro-oscillator mixing that fully vibrates, 37 ℃ of incubations 1 hour.
4) 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.
5) 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.
6) 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 " hepatitis E virus IgM antibody (anti--HAVIgM) chemical luminescence method immune analysis is measured kit " is described meet the calibrating requirement fully.The every index of kit of the present invention all reaches or is higher than the detection index of ELISA reagent.
Embodiment 7 hepatitis E virus IgM antibodies of the present invention (anti--HAVIgM) clinical practice and the ELISA kit of chemical luminescence method immune analysis mensuration kit compare
Clinical hospital use embodiment 1 hepatitis E virus IgM antibody (anti--HAVIgM) chemical luminescence method immune analysis is measured kit and the ELISA kit detects 662 parts of clinical samples, to contrast the coincidence rate that two kinds of kits detect.
One, the source of clinical serum specimen:
Hospital clinical is collected hepatitis E virus patient, non-hepatitis E virus patient's sample and normal human serum sample.
Two, use hepatitis E virus IgM antibody (anti--HAVIgM) chemical luminescence method immune analysis mensuration kit and the comparison of ELISA kit
1, method
Clinical " hepatitis E virus IgM antibody (anti--HAVIgM) chemical luminescence method immune analysis is measured kit " (by its instructions operation) and the ELISA kit (by its instructions operation) of the embodiment of the invention 1 preparation of using respectively detects 662 parts of clinical samples (acute viral hepatitis type E 55 examples, viral hepatitis type E IgG antibody positive 38 examples, healthy people 421 examples, positive blood sample 42 examples of rheumatoid factor, positive 62 examples of hepatitis B, hepatitis A IgM antibody positive 43 examples), to contrast the coincidence rate of two kinds of kits detections.
2, result
The clinical use embodiment 1 of hospital " hepatitis E virus IgM antibody (anti--HAVIgM) chemical luminescence method immune analysis is measured kit " and ELISA kit comparison and detection result (seeing Table 2)
Two kinds of methods of table 2. clinical hospital contrast relatively
Remarks:
Invention reagent detects 1 example and resists-HEVIgG serum sample, and is anti--the HEVIgM positive; Confirm as anti--HEVIgM positive
Contrast agent detects 1 example and resists-HAVIgM serum sample, and is anti--the HEVIgM positive; Confirm as false positive
Contrast agent detects 1 routine acute viral hepatitis type E serum sample, and is anti--the HEVIgM feminine gender; Confirm as false negative
The use of clinical hospital " hepatitis E virus IgM antibody (anti--HAVIgM) chemical luminescence method immune analysis is measured kit " and showing with ELISA kit comparison and detection result: every index of kit of the present invention has has all met or exceeded the ELISA method, and the specificity 100% of this kit, susceptibility 100%; In the clinical testing 1 example resists-HEVIgG positive serum sample anti--HEVIgM positive that reagent of the present invention detects; 1 routine acute HEV sample serum ELISA reagent detects anti--HEVIgM feminine gender, 1 routine resisting-HAVIgM serum specimen ELISA reagent detects the positive.3 parts of serum samples confirm through clinical testing, and 1 example is anti--and HEVIgG positive serum sample results is positive, is to belong to acute infection later stage patient; 1 routine acute HEV sample serum clinical effectiveness is positive, and ELISA reagent result is the gray area sample; 1 routine resisting-HAVIgM serum specimen clinical effectiveness is negative.This kit and rheumatoid factor positive serum, hepatitis B positive serum, hepatitis A IgM positive serum no cross reaction.The hepatitis E virus IgM antibody of invention (anti--HAVIgM) chemical luminescence immune assay determination reagent kit specificity, sensitivity, stability all are higher than the ELISA kit, can be applied to clinical Hepatitis E medical diagnosis on disease.
Claims (10)
1, a kind of hepatitis E virus IgM antibody chemical luminescence immune assay determination reagent kit is characterized in that, kit of the present invention comprises: hepatitis E virus IgM antibody positive and negative reference substance; Bag is by solid phase carrier; The enzyme labeling bond; The chemical luminous substrate liquid that described enzyme acted on; And concentrated cleaning solution.
2, kit of the present invention according to claim 1 is characterized in that, described bag is microwell plate, plastic bead, plastic tube or the magnetic-particle of anti-people-μ chain antibody (monoclonal antibody or how anti-) bag quilt by solid phase carrier.
3, kit of the present invention according to claim 1 is characterized in that, the hepatitis E virus recombinant antigen that described enzyme labeling bond is horseradish peroxidase or alkali phosphatase enzyme mark.
4, kit of the present invention according to claim 1 is characterized in that, described chemical luminous substrate is 1,2-two oxidative ethane analog derivatives, luminol or different luminol.
5, kit of the present invention according to 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: with hepatitis E virus IgM antibody positive and negative serum preparation reference substance; With anti-people-μ chain antibody (monoclonal antibody or how anti-) direct coated solid phase carrier; With enzyme labeling recombined hepatitis E hepatitis virus antigen; The preparation chemical luminous substrate liquid that enzyme acted on; The preparation concentrated cleaning solution;
Chemical luminous substrate liquid and concentrated cleaning solution that hepatitis E virus recombinant antigen, this enzyme of the above-mentioned hepatitis E virus IgM antibody positive and negative of packing reference substance, enzyme labeling acted on; And be assembled into finished product.
7, method as claimed in claim 6, it is characterized in that, described bag is by the step 2 of solid phase carrier) adopt following method preparation: with 0.1M pH value is anti-people-μ chain antibody that 8.0 borate buffer is mixed with desired concn, and coating buffer is carried on the solid phase carrier; With containing 0.1% casein, 1% sucrose, 0.01% gelatin, 0.1%Proclin300, the pH value is 7.0～7.5, concentration is that the phosphate buffer of 0.01M is as the above-mentioned solid phase carrier of confining liquid sealing.
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 the solid phase carrier of anti-people-μ chain antibody (monoclonal antibody or how anti-).
9, method as claimed in claim 6 is characterized in that, described marker enzyme is horseradish peroxidase or alkaline phosphatase.
10, method as claimed in claim 6 is characterized in that, described chemical luminous substrate is 1,2-two oxidative ethane analog derivatives, luminol or different luminol; 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 |
|---|---|---|---|
| CNA2008101032644A CN101551395A (en) | 2008-04-02 | 2008-04-02 | Chemiluminescence immunoassay kit of hepatitis E virus IgM antibody and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008101032644A CN101551395A (en) | 2008-04-02 | 2008-04-02 | Chemiluminescence immunoassay kit of hepatitis E virus IgM antibody and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101551395A true CN101551395A (en) | 2009-10-07 |
Family
ID=41155751
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2008101032644A Pending CN101551395A (en) | 2008-04-02 | 2008-04-02 | Chemiluminescence immunoassay kit of hepatitis E virus IgM antibody and preparation method thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103267846A (en) * | 2013-06-09 | 2013-08-28 | 湖南农业大学 | ELISA (Enzyme Linked Immunosorbent Assay) kit for detecting porcine circovirus 2 (PCV2) IgM antibody |
| CN104090105A (en) * | 2014-07-10 | 2014-10-08 | 广州市丰华生物工程有限公司 | Method and kit for detecting hepatitis E virus (HEV) antibody and method for preparing kit |
| CN105954510A (en) * | 2016-06-30 | 2016-09-21 | 深圳市亚辉龙生物科技股份有限公司 | Chemiluminescence immunodetection kit for anti-beta 2 glycoprotein I antibodies IgG and preparation method of kit |
| CN112433050A (en) * | 2020-11-04 | 2021-03-02 | 上海赛罕生物技术有限公司 | Chlamydia pneumoniae IgM antibody detection kit and detection method thereof |
| CN112986556A (en) * | 2021-05-12 | 2021-06-18 | 佛山微奥云生物技术有限公司 | Chip for rapid detection and processing method thereof |
-
2008
- 2008-04-02 CN CNA2008101032644A patent/CN101551395A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103267846A (en) * | 2013-06-09 | 2013-08-28 | 湖南农业大学 | ELISA (Enzyme Linked Immunosorbent Assay) kit for detecting porcine circovirus 2 (PCV2) IgM antibody |
| CN104090105A (en) * | 2014-07-10 | 2014-10-08 | 广州市丰华生物工程有限公司 | Method and kit for detecting hepatitis E virus (HEV) antibody and method for preparing kit |
| CN105954510A (en) * | 2016-06-30 | 2016-09-21 | 深圳市亚辉龙生物科技股份有限公司 | Chemiluminescence immunodetection kit for anti-beta 2 glycoprotein I antibodies IgG and preparation method of kit |
| CN112433050A (en) * | 2020-11-04 | 2021-03-02 | 上海赛罕生物技术有限公司 | Chlamydia pneumoniae IgM antibody detection kit and detection method thereof |
| CN112986556A (en) * | 2021-05-12 | 2021-06-18 | 佛山微奥云生物技术有限公司 | Chip for rapid detection and processing method thereof |
| CN112986556B (en) * | 2021-05-12 | 2022-02-18 | 佛山微奥云生物技术有限公司 | Chip for rapid detection and processing method thereof |
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