CN103048456A - Hepatitis B virus PreS1 antigen enzyme-linked immunoassay kit employing one-step method - Google Patents
Hepatitis B virus PreS1 antigen enzyme-linked immunoassay kit employing one-step method Download PDFInfo
- Publication number
- CN103048456A CN103048456A CN2012103769647A CN201210376964A CN103048456A CN 103048456 A CN103048456 A CN 103048456A CN 2012103769647 A CN2012103769647 A CN 2012103769647A CN 201210376964 A CN201210376964 A CN 201210376964A CN 103048456 A CN103048456 A CN 103048456A
- Authority
- CN
- China
- Prior art keywords
- antibody
- pres1
- bottle
- preparation
- hepatitis
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The invention provides a hepatitis B virus PreS1 antigen enzyme-linked immunoassay kit employing a one-step method. The concentration of HBs-Ag-PreS1 protein in blood serum of a sufferer can be exclusively detected. The kit comprises an elisa plate enveloped reaction strip, an enzyme labeling pre-S1 antibody, a positive control solution, a negative control solution, a bottle of 20-fold wash buffer, a bottle of substrate buffer A, a bottle of substrate buffer B and a bottle of stop buffer (2NH2SO4), wherein the elisa plate enveloped reaction strip is prepared from a primary ingredient avidin which is enveloped in advance and then added to a biotin anti-hepatitis B virus PreS1 monoclonal antibody or an anti-HBS antibody. The kit is good in uniformity, specificity and sensitivity, rapid and convenient to collect, can obtain an experiment result about 30 minutes by the one-step method, and is time-saving (the experiment can be fastest finished by 6-8 hours in a PCR (polymerase chain reaction) method) compared with the PCR detection method. The positivity detected by clinical PreS1 protein and the HBV-DNA positivity detected by PCR have good coincidence rate.
Description
Technical field
The invention belongs to the biologic product technology field.Be specifically related to a kind of single stage method hepatitis B virus pro-S 1 (PreS1) antigen (Ag) ELISA measuring reagent kit and preparation method.
Background technology
China belongs to the hepatitis type B virus district occurred frequently, hepatitis B has become public's property problem of harm China people's health, the conventional project that pattern of hepatitis B virus serologic markers (HBVM) is diagnosed as hepatitis type B virus can not satisfy clinical, the evidence that HBV-DNA, viral surface antigen (HBsAg), hepatitis B virus pro-S 1 (PreS1Ag) antigen all infect and copy in human body as HBV.
HBV is hepadnavirus, is comprised of an incomplete double-stranded DNA, about 3200 amino acid, and it is S district, C district, P district, X district that 4 opening code-reading frames are arranged.The S gene regions can be divided into again three sections, begins to be followed successively by front S1 (preS1) district, front S2 (preS2) district and S district from the upstream.PreS1 is former to have stronger immunogenicity, is comprised of 108 or 119 amino acid.HBV in the past S1 district amino acid 21-47 fragment as binding site in conjunction with liver cell, as long as this fragment intact just be infectious and the height immunogenicity.PreS1 antigen appears at the very early time of acute hepatitis b virus infection, and is more Zao than the HbeAg appearance, is early stage communicable sign.The massive duplication of hepatitis type B virus, cause a large amount of PreS1Ag to express, stimulate body to produce stronger immune response, thereby cause a large amount of hepatocellular injuries, and think the coat protein composition of PreS1 as HBV, play a part key in the process of HBV host cells infected, most important mediation position is amino (AA) the 21-47 fragment of pre-s1 protein, and the virus of variation just is infectious as long as this section is intact.
PreS1 is in the whole cycle of virus infections body, the unique effect of PreS1, can react more fully the humoral immunity of body, until the process of turning out cloudy of the course of disease, to turn be the earliest sign of virus sweep to PreS1Ag the moon during such as oxyhepatitis, otherwise, the PreS1Ag lasting masculin, oxyhepatitis will be developed to chronic hepatitis.Therefore, PreS1Ag and HBV-DNA have good consistance at the reflection hepatitis B virus duplication.PreS1Ag reflects the situation of hbv replication more sensitively than HBeAg.
But generally use at present double antibody sandwich method to detect value of HBV PreS 1 antigen, has good specificity, can exist the monoclonal antibody use amount large on the ELISA Plate but utilize merely unable absorption monoclonal antibody or polyclonal antibody to be adsorbed on, loss of activity is large, the shortcoming such as particularly betweenrun precision is poor, and Avidin is coated onboard, add again biotinylated antibody, then can well address this problem, it not only can reduce difference between batch, and can be in testing process can directly add sample serum and enzymic-labelled antibody a step and react, form Solid-phase avidin-anti-PreS1 antigen antibody complex of biotin words antibody-value of HBV PreS 1 antigen-enzyme mark, then add substrate buffer solution first and second and react, the content of the value of HBV PreS 1 antigen in just can quantitative test serum.
Summary of the invention
Technical matters to be solved by this invention is to solve the problem of traditional ELISA Plate difference between batch deficiency, has designed and set up the detection kit of single stage method PreS1 antigen.
The invention provides a kind of " single stage method PreS1 antigen ELISA measuring reagent kit ", this kit is to comprise the in advance coated rear coated reaction of ELISA Plate bar, enzyme labeling former S 1 immune body, one of positive control solution, 1 bottle of one, 20 times dense cleansing solution of negative controls, 1 bottle of substrate buffer solution first, 1 bottle of substrate buffer solution second and 1 bottle of composition of stop buffer (2N H2SO4) that adds biotinylation anti-hepatitis virus PreS1 monoclonal antibody or anti-HBS antibody of principal ingredient Avidin.
1, antibody preparation:
(1) utilize HBV-DNA restructuring PreS1 Ag genetic fragment to prepare antigen by prokaryotic expression or eukaryotic expression;
(2) preparation of Dispersal risk and purifying: with above-mentioned recombinant antigen immune guinea pig, get anti-PreS1 antiserum or immune Balb/C mouse gets positive mice spleen cell and myeloma; Fusion of Cells becomes hybridoma, and screening positive clone is built strain, gets ascites; Antiserum or ascites with saltout, ion-exchange chromatography or affinity chromatography carry out purifying, gets pure anti-PreS1 antibody, its purity is more than 95%.
(3) with Avidin elder generation coated elisa plate, and add biotin labeled anti-hepatitis virus PreS1 monoclonal antibody or anti-HBS carries out the coated reaction of secondary, make the pre-coated reaction bar of antibody 48-96 hole;
(4) the anti-hepatitis virus PreS1 antibody ELISA of horseradish peroxidase (HRP) and purifying is made enzyme labelled antibody;
(5) preparation positive control solution, negative controls;
(6) 20 times of concentrated washing lotions of preparation;
(7) preparation substrate buffer solution first;
(8) preparation substrate buffer solution second;
(9) preparation stop buffer (2N H2SO4).
(10) packing mentioned reagent box inter prediction is divided in bottle or the conical centrifuge tube on demand by all the other seven kinds of compositions of plate;
(11) specificity, sensitivity, accuracy, the qualified stability of calibrating kit;
(12) be assembled into finished product.
2, kit of the present invention can followingly operate in use:
(1) takes out coated cylindrical void or plate, return to room temperature.
(2) dosing: with concentrated cleaning solution (20X) with distilled water or deionized water diluted for use (20 times of dense cleansing solution 1ml+19ml distilled water are working fluid).
(3) application of sample: every hole adds serum 50ul to be checked, and each plate is established positive control one hole (50ul), negative control one hole (50ul), blank one hole (adding distil water 50ul), and then except the blank hole, the enzyme-added marking fluid 50ul in each hole,
(4) incubation: use the rearmounted 37 ℃ of incubations of shrouding film shrouding 30 minutes.
(5) washing: carefully take the shrouding film off, liquid in the hole is dried, fully wash 5 times with cleansing solution, button is done.
(6) every hole adds nitrite ion A, B each 1 (50ul), the mixing that vibrates gently, and 37 ℃ of lucifuges developed the color 15 minutes
(7) measure the OD value: every hole adds 1 of stop buffer (50ul), and the mixing that vibrates is gently set the enzyme non-analysis meter ripple and is longer than 450nm place (suggestion detects with dual wavelength 450/630nm), measures each hole OD value.
(8) result judges
Negative control (A value) * 2.5=critical value (cut off value), negative control are higher than at 0.05 o'clock to be calculated by actual OD value, and negative control is pressed 0.05 less than 0.05 and calculated, and all specimen hole A values to be checked are namely positive greater than critical value.
Kit of the present invention can detect the concentration of HBsAg-PreS1 albumen among the patients serum very single-mindedly.It has the characteristics such as easy, sensitive and stable.And this kit is easy and simple to handle fast, adopt single stage method about 30 minutes, to obtain experimental result, than PCR detection method save time (PCR method need the soonest 6-8 hour finish experiment), through the positive HBV-DNA positive that detects with PCR of clinic trial PreS1 Protein Detection good coincidence rate is arranged, this kit is compared with the PCR method, PCR needs valuable instrument and equipment, expendable reagent is expensive, the charge valency is high again, and need not valuable instrument and equipment in the whole experiment of this kit, expendable reagent is cheap and the charge valency is cheap, can be applicable to situation of all-level hospitals and clinical trial center, also can be used for the epidemiology generaI investigation.Single stage method PreS1 Ag kit has advantage easy, sensitive, good reproducibility, can detect complete HBV, can be commonly used.
Embodiment
Embodiment 1
1, preparation single stage method PreS1 Ag enzyme is exempted from the production stage of kit:
1, biotinylated antibody preparation:
(1) purifying coated antibody (HBsAb): use the recombinant antigen immune guinea pig, get anti-PreS1 antiserum or immune Balb/C mouse gets positive mice spleen cell and the myeloma cell is fused into hybridoma, screening positive clone is built strain, gets ascites; Antiserum or ascites with saltout, ion-exchange chromatography or affinity chromatography carry out purifying, gets pure anti-PreS1 antibody, its purity is more than 95%.Tire greater than 100,000.
(2) the anti-PreS1 antibody of biotin coupling:
With conventional method biotin (BNHS) is dissolved in N, N-dimethylformamide (DMF) is made into 1mg/Ml, being 9.0 NaHCO3 with 0.1mol/L pH value becomes 1-2mg/mL with the anti-PreS1 antibody dilution of purifying, be that 1: 8 or weight ratio are to mix in about 1: 7 by BNHS and the volumetric ratio of antibody, react 2-4h under the stirring at room, packing into dialysis is 7.2 PBS to the 0.05mol/LpH value, 4 ℃ of dialysed overnight, bond adds isopyknic glycerine, in a small amount packing, and-20 ℃ once save backup.
2, enzymic-labelled antibody preparation:
(1) antibody preparation:
The PreS1 Ag immune guinea pig of genetic recombination must resist the PreS1 Ag immunity Balb/C mouse of PreS1 antibody or restructuring to get ascites, antiserum or ascites with saltout, ion-exchange chromatography or affinity chromatography carry out purifying, get pure anti-PreS1 antibody, its purity is more than 95%.Tire greater than 100,000.
(2) enzymic-labelled antibody preparation
Anti-PreS1 antibody sodium periodate method and horseradish peroxidase coupling
(3) enzymic-labelled antibody concentration is selected
Adopt the square formation titrimetry to select the working concentration of enzymic-labelled antibody greater than 1: 2000.
The PreS1 recombinant antigen dilutes from the 10ug/ml multiplication, coated elisa plate, and the enzyme labelled antibody that adds gradient dilution is measured, to determine optimal dilution.
3, the preparation of pre-coated antibody panel
(1) coated
Be the Avidin coating buffer of the citrate buffer solution preparation desired concn of 4.5-5.0 with the pH value of 0.05M, coating buffer added the solid phase carrier enzyme put on, put in the wet box and add a cover, the 40C drying of spending the night.
(2) washing
With physiological saline washing three times, to remove the residue Avidin.
(3) sealing
Adjust PH to 7.2, confining liquid 300 μ L are added in each hole of microwell plate, leave standstill afterwards drying in 5 seconds, the aforesaid operations triplicate gets rid of confining liquid, pats dry.
(4) secondary reaction is coated
Biotinylated anti-PreS1 antibody is diluted to suitable concn with the PBS of 0.02M, adds 110 μ L in each hole of microwell plate, put in the wet box and add a cover, the 40C drying of spending the night.PBS solution washing three times and dehumidifier with 0.02M are dry, have put into the drying agent aluminium foil bag and have sealed up for safekeeping for subsequent use.
4, the preparation of positive control
The Serum of Patients with Hepatitis B that HBeAg and HBV-DNA are positive simultaneously.600C placed 1 hour, and aseptic filtration is measured A value＞0.3 with this medicine box, and is for subsequent use, packing.
5, the preparation of negative control
At 0-0.03, add 2/10000ths thimerosals with this kit measurement normal human serum A value, packing is for subsequent use.
6, enzyme mark monoclonal antibody configuration
With containing 10% calf serum and 90%0.15MPBS dilution Anti-presl-HRP, dilute 20 times, packing.
7, enzyme mark monoclonal antibody dilution
8, substrate solution A
9, substrate solution B
Na2HPO4.12H2O 1.7g
Citric acid .H2O 0.5g
Redistilled water 100ml
After adjusting pH to 5.0, add the solution 25 μ l that 10mlDMSO contains 60mgTMB
10, stop buffer
Dense H2SO4 10ml
Redistilled water 80ml
11,20X cleansing solution
12, the composition of semi-manufacture and finished product
Above-mentioned (one) → (11) step products obtained therefrom is packed in bottle and the conical centrifuge tube, is semi-manufacture.Extract three parts of process specificitys, stability, sensitivity and precision assay approvals out and just can be assembled into the presl kit.Be assembled into and also need extract three parts behind the box out and equally could sell through assay approval with semi-manufacture.
Embodiment 2
Single stage method PreS1 Ag enzyme is exempted from the operation steps of kit
1, operation steps is as follows:
(1) takes out coated cylindrical void or plate, return to room temperature.
(2) dosing: with concentrated cleaning solution (20X) with distilled water or deionized water diluted for use (20 times of dense cleansing solution 1ml+19ml distilled water are working fluid).
(3) application of sample: every hole adds serum 50ul to be checked, and each plate is established positive control one hole (50ul), negative control one hole (50ul), blank one hole (adding distil water 50ul), and then except the blank hole, the enzyme-added marking fluid 50ul in each hole.
(4) incubation: use the rearmounted 37 ℃ of incubations of shrouding film shrouding 30 minutes.
(5) washing: carefully take the shrouding film off, liquid in the hole is dried, fully wash 5 times with cleansing solution, button is done.
(6) every hole adds nitrite ion A, B each 1 (50ul), the mixing that vibrates gently, and 37 ℃ of lucifuges developed the color 15 minutes
(7) measure the OD value: every hole adds 1 of stop buffer (50ul), and the mixing that vibrates is gently set the enzyme non-analysis meter ripple and is longer than 450nm place (suggestion detects with dual wavelength 450/630nm), measures each hole OD value.
(8) result judges
Negative control (A value) * 2.5=critical value (cut off value), negative control are higher than at 0.05 o'clock to be calculated by actual OD value, and negative control is pressed 0.05 less than 0.05 and calculated, and all specimen hole A values to be checked are namely positive greater than critical value.
Claims (5)
1. one kind " single stage method value of HBV PreS 1 antigen ELISA measuring reagent kit " is characterized in that this kit comprises:
(1) with Avidin elder generation coated elisa plate, and adds biotin labeled front S1 monoclonal antibody or anti-HBS carries out the coated reaction of secondary, make the pre-coated reaction bar of antibody 48-96 hole;
(2) enzyme labeling former S 1 immune body;
(3) 1 of positive control solution, 1 of negative controls;
1 bottle of (4) 20 times of concentrated washing lotion;
(5) the substrate buffer solution first is 1 bottle;
(6) substrate buffer solution second is 1 bottle;
(7) 1 bottle of composition of stop buffer (4N H2SO4).
2. the preparation method such as right 1 described a kind of " single stage method value of HBV PreS 1 antigen ELISA measuring reagent kit " is characterized in that the method comprises the following steps:
(1) utilize HBV-DNA restructuring PreS1 Ag genetic fragment to prepare antigen by prokaryotic expression or eukaryotic expression;
(2) preparation of Dispersal risk and purifying: with above-mentioned recombinant antigen immune guinea pig, get anti-PreS1 antiserum or immune Balb/C mouse gets positive mice spleen cell and the myeloma cell is fused into hybridoma, screening positive clone is built strain, gets ascites; Antiserum or ascites with saltout, ion-exchange chromatography or affinity chromatography carry out purifying, gets pure anti-PreS1 antibody, its purity is tired greater than 100,000 more than 95%.
(3) with Avidin elder generation coated elisa plate, and add biotin labeled anti-hepatitis virus PreS1 monoclonal antibody or anti-HBS carries out the coated reaction of secondary, make the pre-coated reaction bar of antibody 48-96 hole;
(4) the anti-hepatitis virus PreS1 antibody ELISA of horseradish peroxidase (HRP) and purifying is made enzyme labelled antibody;
(5) preparation positive control solution, negative controls;
(6) 20 times of concentrated washing lotions of preparation;
(7) preparation substrate buffer solution first;
(8) preparation substrate buffer solution second;
(9) preparation stop buffer (2N H2SO4).
(10) packing mentioned reagent box inter prediction is divided in bottle or the conical centrifuge tube on demand by all the other seven kinds of compositions of plate;
(11) specificity, sensitivity, accuracy, the qualified stability of calibrating kit;
(12) be assembled into finished product.
3. according to right 1 described a kind of " single stage method value of HBV PreS 1 antigen ELISA measuring reagent kit ", it is characterized in that the pre-coated reaction bar of wherein said antibody 48-96 hole, the front antibody of enzyme labeling, positive control solution are HBV-DNA and the equal positive serum of HBeAg, negative controls is that normal human serum, dense cleansing solution are phosphoric acid-Tween-20 damping fluid, the substrate buffer solution first is that H2O2, substrate buffer solution second are that 3.3 ', 5.5 '-tetramethyl benzidine and stop buffer are (4N H2SO4).
4. method as claimed in claim 3, it is characterized in that, the coated solid phase carrier ELISA Plate of described Avidin adopts method: the pH value with 0.05M is the Avidin coating buffer of the citrate buffer solution preparation desired concn of 4.5-5.0, coating buffer is added the solid phase carrier enzyme to be put on, and wash, seal, add again after biotinylated hepatitis B virus Pres 1 monoclonal antibody or anti-HBS antibody reacts, after washing, dehumidifier drying, seal with aluminium foil bag.
5. method as claimed in claim 2 is characterized in that described thickening and washing formula of liquid is: contain 0.02MPBS, 8.9%NaCl and 0.5%Tween-20.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103769647A CN103048456A (en) | 2012-10-08 | 2012-10-08 | Hepatitis B virus PreS1 antigen enzyme-linked immunoassay kit employing one-step method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012103769647A CN103048456A (en) | 2012-10-08 | 2012-10-08 | Hepatitis B virus PreS1 antigen enzyme-linked immunoassay kit employing one-step method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103048456A true CN103048456A (en) | 2013-04-17 |
Family
ID=48061170
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012103769647A Pending CN103048456A (en) | 2012-10-08 | 2012-10-08 | Hepatitis B virus PreS1 antigen enzyme-linked immunoassay kit employing one-step method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103048456A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105842447A (en) * | 2016-03-18 | 2016-08-10 | 南昌大学 | Detection method for hepatitis B surface antigen |
| CN107085111A (en) * | 2017-04-14 | 2017-08-22 | 江苏福隆生物技术有限公司 | The board-like chemoluminescence method detection kit and preparation method of hepatitis B virus pre S 1 antigen |
| CN107167590A (en) * | 2017-04-14 | 2017-09-15 | 江苏福隆生物技术有限公司 | The board-like chemoluminescence method detection kit and preparation method of carbohydrate antigen 242 |
| CN107462721A (en) * | 2017-07-10 | 2017-12-12 | 江苏福隆生物技术有限公司 | Board-like the chemoluminescence method detection kit and preparation method of cytokeratin 19 fragment |
| CN107490695A (en) * | 2017-07-10 | 2017-12-19 | 江苏福隆生物技术有限公司 | Board-like the chemoluminescence method detection kit and preparation method of CA50 |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1510421A (en) * | 2002-12-23 | 2004-07-07 | 杜凤鸣 | Hepatitis B virus fore S1 antigen enzyme linked immunoassay reagent box and preparation thereof |
| CN1512178A (en) * | 2002-12-27 | 2004-07-14 | 杜凤鸣 | Detection reagent kid for hepatitis B virus proantigen S1 enzyme-linked immunology by quantitative method and its preparing method |
| CN1869701A (en) * | 2006-06-27 | 2006-11-29 | 厦门大学 | Method for joint investigating hepatitis B virus pro S1 antigen and nuclear antigen and diagnostic kit |
| CN101377506A (en) * | 2008-04-16 | 2009-03-04 | 北京科美东雅生物技术有限公司 | Monophosphoinositide proteoglycans-3 chemiluminescence immune analysis determination reagent kit and preparing method thereof |
| CN101382553A (en) * | 2007-10-25 | 2009-03-11 | 北京科美东雅生物技术有限公司 | Large protein pre-S surface antigen for hepatitis B virus chemiluminescence immune assay kit and method for making same |
| CN101692087A (en) * | 2009-09-30 | 2010-04-07 | 暨南大学 | Method for preparing streptavidin pre-coated elisa plate and application |
-
2012
- 2012-10-08 CN CN2012103769647A patent/CN103048456A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1510421A (en) * | 2002-12-23 | 2004-07-07 | 杜凤鸣 | Hepatitis B virus fore S1 antigen enzyme linked immunoassay reagent box and preparation thereof |
| CN1512178A (en) * | 2002-12-27 | 2004-07-14 | 杜凤鸣 | Detection reagent kid for hepatitis B virus proantigen S1 enzyme-linked immunology by quantitative method and its preparing method |
| CN1869701A (en) * | 2006-06-27 | 2006-11-29 | 厦门大学 | Method for joint investigating hepatitis B virus pro S1 antigen and nuclear antigen and diagnostic kit |
| CN101382553A (en) * | 2007-10-25 | 2009-03-11 | 北京科美东雅生物技术有限公司 | Large protein pre-S surface antigen for hepatitis B virus chemiluminescence immune assay kit and method for making same |
| CN101377506A (en) * | 2008-04-16 | 2009-03-04 | 北京科美东雅生物技术有限公司 | Monophosphoinositide proteoglycans-3 chemiluminescence immune analysis determination reagent kit and preparing method thereof |
| CN101692087A (en) * | 2009-09-30 | 2010-04-07 | 暨南大学 | Method for preparing streptavidin pre-coated elisa plate and application |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105842447A (en) * | 2016-03-18 | 2016-08-10 | 南昌大学 | Detection method for hepatitis B surface antigen |
| CN107085111A (en) * | 2017-04-14 | 2017-08-22 | 江苏福隆生物技术有限公司 | The board-like chemoluminescence method detection kit and preparation method of hepatitis B virus pre S 1 antigen |
| CN107167590A (en) * | 2017-04-14 | 2017-09-15 | 江苏福隆生物技术有限公司 | The board-like chemoluminescence method detection kit and preparation method of carbohydrate antigen 242 |
| CN107085111B (en) * | 2017-04-14 | 2019-02-01 | 江苏福隆生物技术有限公司 | The board-like chemoluminescence method detection kit and preparation method of hepatitis B virus pre S 1 antigen |
| CN107462721A (en) * | 2017-07-10 | 2017-12-12 | 江苏福隆生物技术有限公司 | Board-like the chemoluminescence method detection kit and preparation method of cytokeratin 19 fragment |
| CN107490695A (en) * | 2017-07-10 | 2017-12-19 | 江苏福隆生物技术有限公司 | Board-like the chemoluminescence method detection kit and preparation method of CA50 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102798725B (en) | Diagnostic kit for determination of serum total IgE, preparation method and application method | |
| ES2312062T3 (en) | PROCEDURE FOR THE DETECTION OF PROBNP N-TERMINAL. | |
| CN103364568B (en) | Laminin nano-magnetic particle chemiluminescent immunity quantitative detection kit and preparation method thereof | |
| CN103048456A (en) | Hepatitis B virus PreS1 antigen enzyme-linked immunoassay kit employing one-step method | |
| CN100582780C (en) | Micro-molecule indirectly labeled dual-antigen sandwich method for determining hepatitis C virus total antibody | |
| CN107462726B (en) | Dual quantitative ELISA detection method is immunized in a kind of magnetic enzyme sandwich based on double monoclonal antibodies | |
| CN101363853A (en) | Human immunodeficiency virus antigen/antibody chemiluminescence immune assay determination kit and method for preparing same | |
| CN101832999A (en) | Antigen antibody complex dissociation liquid kit and application thereof | |
| CN101377505A (en) | Des-gamma-carboxy-pro-thrombin microplate chemiluminescence immune analysis determination reagent kit and preparing method thereof | |
| JP2001124775A (en) | Flow-through assay for visually detecting presence of influenza type a and type b | |
| CN103175963A (en) | Treponema (TP) antibody detection method and detection kit thereof | |
| CN106198997B (en) | A kind of kit of Quantitative detection troponin and creatine kinase isozyme | |
| US4929543A (en) | Process for the determination of an antibody in human body fluids | |
| CN102768274A (en) | Human immunodeficiency virus P24 antigen enzyme-linked immunodetection kit | |
| CN105445463A (en) | Hepatitis E virus IgG antibody detection kit and preparation method and application thereof | |
| CN115176162A (en) | Novel coronavirus antigen and detection use thereof | |
| CN101498730A (en) | Improved double-antigen sandwiching immunity detection method | |
| CN101614737A (en) | A kind of colloidal gold strip and its production and application | |
| CN104076146A (en) | ELISA (Enzyme-Linked Immunosorbent Assay) kit for detecting human anti-rabies virus antibody | |
| JP2524101B2 (en) | Immunodetermination method for detection of common determinant antibody against hepatitis B | |
| CN101598730A (en) | Hepatitis B virus E antibody through dual-antigen sandwich method external diagnosis reagent case and preparation method | |
| CN102368068B (en) | Kit for detecting chlamydia pneumoniae IgM antibody | |
| CN104297482B (en) | Epstein-Barr virus VCA/NA1-IgA antibody combined detection reagent and preparation method thereof | |
| CN101523216A (en) | Method of detecting antibodies against a series of human immunodeficiency virus proteins | |
| CN111458498A (en) | Hand-foot-mouth EV71 antigen detection kit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20130417 |