CN100582780C - Micro-molecule indirectly labeled dual-antigen sandwich method for determining hepatitis C virus total antibody - Google Patents
Micro-molecule indirectly labeled dual-antigen sandwich method for determining hepatitis C virus total antibody Download PDFInfo
- Publication number
- CN100582780C CN100582780C CN200610027230A CN200610027230A CN100582780C CN 100582780 C CN100582780 C CN 100582780C CN 200610027230 A CN200610027230 A CN 200610027230A CN 200610027230 A CN200610027230 A CN 200610027230A CN 100582780 C CN100582780 C CN 100582780C
- Authority
- CN
- China
- Prior art keywords
- hepatitis
- hcv
- antigen
- antibody
- micromolecule
- 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.)
- Active
Links
Abstract
The invention relates to a method for testing the total antibody of hepatitis C virus (HCV), based on dual-antibody interlayer method of small-molecule indirect mark, wherein said method is characterized in that: it uses small molecule matter as mark; uses dual-antigen interlayer method to test the total antibody of sample; and said method comprises: using small molecule material to mark HCV antigen; using HCV antigen to pack solid material; using single generate material to mark small molecule antibody or other material that combined with small molecule; the HCV antibody of sample can be reacted with HCV antigen of solid material surface and small molecule mark, to form dual-antigen interlayer composite; then reacting the small molecule with single report molecule, to make the composite with detected signal generate material. The invention can keep activity of antigen, with signal amplifying function and better specificity of dual-antigen interlayer mode, while it can detect variable kinds of antibodies, to improve the detecting sensitivity and specificity.
Description
One, technical field:
The present invention relates to external immunodiagnosis field, particularly a kind of method based on the total antibody of micro-molecule indirectly labeled double antigens sandwich immunoassays hepatitis C virus (HCV).
Two, background technology:
Hepatitis C is a kind of worldwide disease, about 3% (1.7 hundred million people) of global HCV infection rate; China's hepatitis C prevalence rate is 3.2%, and number of the infected surpasses 4,100 ten thousand.Because most of HCV acute infection persons do not have symptom or symptom not obvious, and the body inner virus is difficult to remove, about 70～80% case will transfer chronic HCV infection to, chronic HCV infection can cause necrosis of liver chronic inflammation and fiberization, part patient can develop into cirrhosis even hepatocellular carcinoma (HCC), and is very harmful to patient's health and lives.HCV infects has become serious society and public health problem.
Blood-borne is the main path that HCV infects; The HCV detection of antibodies has reduced the blood transfusion of HCV effectively and has infected, and also the diagnosis for hepatitis C provides valuable information.The indirect method that HCV antibody all adopts the mark second antibody is detected in domestic each hospital, blood station at present, and false positive rate is very high; For reducing the too much false positive of indirect method, the Chinese patent of publication number CN1548958A (Li Chenyang, Li Yulin.Antibody of HCV bridge-type double antigens sandwich ELISA detection method.Open day: on November 24th, 2004, publication number: CN1548958A) disclose a kind of method of utilizing gene recombination technology mark HCV antigen: obtain a kind of fusion by gene expression, this fusion is except containing HCV antigen, also contain one " pontin protein " as the iron thioredoxin, HCV antigen can be connected with anti-" pontin protein " antibody of an enzyme labeling by " pontin protein ", realizes the enzyme labeling of HCV antigen.The Chinese patent of publication number CN1670532A (Wang Baojun, Feng Changfang, Ma Xiaohui.Detect hepatitis C virus antibody diagnosing reagent kit and preparation method thereof, detection method.Open day: on September 21st, 2005, publication number: CN1670532A) also disclose the method for a kind of direct use enzyme molecule by chemical linkage flag HCV antigen, the ELISA that is used for HCV antigen/antibody combination detected.Above-mentioned patent content all adopts bigger protein of molecular weight or polypeptide to serve as a mark.
At home and abroad there is no at present with the micromolecule is the report that the technology of the total antibody of dual-antigen sandwich method for determining HCV of indirect labelling is used to detect the total antibody of human serum HCV.
Three, summary of the invention:
The objective of the invention is, be the double antigens sandwich immunologic detection method that indirect labelling is set up the total antibody of a kind of mensuration hepatitis C virus (HCV) with the micromolecule.This detection method has overcome the shortcoming of big molecule (as: enzyme) mark HCV antigen, can detect IgG, IgM, multiclass HCV antibody such as IgA, IgE in the sample simultaneously, and sample need not dilution, has significantly improved the specificity and the sensitivity of HCV antibody test.
Technical scheme of the present invention: micro-molecule indirectly labeled dual-antigen sandwich method for determining hepatitis C virus total antibody is characterized in that method may further comprise the steps:
A) with small-molecule substance mark HCV antigen;
B) with the antigen coated solid phase material of HCV;
C) generate the anti-small molecular antibody of material mark or other materials that can combine with signal, be prepared into the signal reporter molecules with micromolecule;
D) utilize HCV antibody generation immune response in the HCV antigen of the HCV antigen on solid phase material surface and micromolecule mark and the sample, form the double antigens sandwich compound: HCV antigen (1)-HCV antibody-HCV antigen (2)-micromolecule, utilize signal reporter molecules and micromolecular association reaction then, make the compound of formation have detectable signal generation material, be used for detecting the content of the total antibody of sample HCV.
Described " micromolecule " refers to:
(1) biotin molecule;
(2) other molecular weight is less than 10000 micromolecule haptens, as digoxin, fluorescein, thyroxine, estradiol, estriol, triiodo thryonine, testosterone, progesterone etc.
Described signal generates material and is meant enzyme, fluorescent material, rare earth ion such as Eu
3+, Sm
3+, Tb
3+, Dy
3+And chelate aglucon, chemiluminescent substance such as bifurcation pyridine ester and derivant thereof, tris (bipyridine) ruthenium.
Described signal reporter molecules is meant by signal and generates material molecule mark, that can characterize specificity association reaction degree, as the streptavidin of anti-small molecular antibody, enzyme labeling or the luminescent substance mark of rare earth ion mark.
Described immune response refers to antigen or haptens and combines at the specificity between their antibody, as the micromolecule haptens among the present invention with combine at the specificity between their antibody.
Described double antigens sandwich compound be with HCV antibody for " bridge ", HCV antigen (1)-HCV antibody-HCV antigen (the 2)-micromolecule compound that forms by bridging;
Among the present invention be the method that indirect labelling is measured the double antigens sandwich immunity of the total antibody of HCV, it is characterized in that with the micromolecule:
(1) method of usefulness small-molecule substance mark HCV antigen:
Directly use micromolecule and the HCV antigen-reactive that is connected with reactive functional; Or introduce reactive functional at micromolecule or HCV antigen by activation method, again with HCV antigen or micromolecule reaction; Also can directly on recombinant HCV antigen, introduce micromolecule, finish the mark of recombinant HCV antigen by zymochemistry and gene recombination technology;
(2) with the method for the antigen coated solid phase material of HCV:
HCV antigen is dissolved in damping fluid, contacts and leave standstill 5～24 hours with solid phase material; With the damping fluid washing solid phase material that contains surfactant; Add the damping fluid of casein containing protein, left standstill 3～12 hours; Discard the damping fluid of casein containing protein, dry solid phase material.Finish the preparation of the antigen coated solid phase material of HCV.
(3) preparation method of signal reporter molecules that is: generates the method for the anti-small molecular antibody of material mark or other materials that can combine with micromolecule with enzyme, fluorescent material, chemiluminescent substance or other signal:
Anti-small molecular antibody or other materials that can combine with micromolecule are combined by sodium periodate method and enzyme; Anti-small molecular antibody or other materials that can combine with micromolecule are mixed with fluorescent material that has reactive functionality or chemiluminescent substance, they are combined.In the presence of difunctional functional group, anti-small molecular antibody or other materials that can combine with micromolecule are mixed with enzyme, fluorescent material or chemiluminescent substance, they are combined.
Among the present invention be the method that indirect labelling is measured the double antigens sandwich immunity of the total antibody of HCV, it is characterized in that the total detection of antibodies of HCV may further comprise the steps in the described detection sample of step d) with the micromolecule:
(1) sample of adding not diluted in solid phase material;
(2) add micromolecule mark HCV antigen; Utilize the immune response of HCV antibody in the HCV antigen of the HCV antigen on solid phase material surface and micromolecule mark and the sample, form the double antigens sandwich compound: HCV antigen (1)-HCV antibody-HCV antigen (2)-micromolecule;
(3) add the signal reporter molecules, the signal reporter molecules combines with micromolecule, forms to have the detected compound that signal generates material;
(4) strong and weak yin and yang attribute or the antibody titer of determining the total antibody of HCV in the sample of signal that is produced according to the signal reporter molecules.
The present invention adopts micro-molecule indirectly labeled, detects HCV antigen/antibody combination in conjunction with dual-antigen sandwich method, compares with protein or polypeptide marker HCV antigen, and micro-molecule indirectly labeled pattern will significantly be improved the sensitivity and the specificity of HCV antibody test.
With micromolecule mark HCV Detection of antigen HCV antibody, help fully to keep the immune response activity of HCV antigen.Be used for the various HCV antigens of HCV antibody test, with the obvious advantage in the use by the chimeric HCV gene recombinant antigens that forms of a plurality of dominant antigen epi-positions: it had both concentrated high conservative in the HCV albumen, strong immunogenicity, antibody appearance each dominant antigen epi-position early, removed again and the incoherent amino acid sequence of antibodies, therefore good sensitivity and specificity have been arranged; But for this amino acid sequence that concentrates the dominant antigen epi-position, have only the micromolecule of use to be mark, could effectively avoid the decline and the forfeiture of the epitope activity that reasons such as sterically hindered, covering cause, the immune response activity of showing HCV antigen better helps the highly sensitive detection of HCV antibody.In addition, be indirect labelling with the micromolecule, by combining of a plurality of micromolecule on the HCV antigen and signal reporter molecules, can introduce a signal amplifying system, can effectively improve the detection sensitivity of HCV antigen/antibody combination.
It with the micromolecule specificity that the dual-antigen sandwich method of indirect labelling can also significantly improve the HCV antibody test.Except the specific recognition ability of solid phase antigen antagonist, replace the mark second antibody in the indirect method to increase by two important specific factors with the micromolecule labelled antigen: 1) human antibody of non-special absorption or non-specific bond is not discerned by the micromolecule labelled antigen; 2) even envelope antigen exists and nonspecific antibody combining to a certain degree, also can match with it, thereby eliminate the ability of this antibody while in conjunction with envelope antigen and labelled antigen by selecting the another kind of labelled antigen that lacks this non-specific bond.
Owing to the dual-antigen sandwich method that with the micromolecule is indirect labelling has good detection specificity, thereby can use the serum of not diluted or blood plasma as sample (HCV antigen/antibody combination in the sample is not diluted), help detecting of multiclass HCV antigen/antibody combination in the sample (as: IgG, IgM, IgA, IgE etc.).
Based on above-mentioned factor, with the micromolecule sensitivity and the specificity that the total antibody of dual-antigen sandwich method for determining HCV of indirect labelling can effectively improve the HCV antibody test.
Advantage of the present invention or effect are as follows:
(1) with micromolecule mark HCV Detection of antigen HCV antibody, help fully to keep the immune response activity of HCV antigen, effectively improve the sensitivity of HCV detection of antibodies.
(2) can make with micromolecule mark HCV antigen and have a plurality of micromolecule marks on the HCV antigen, thereby can combine, produce effective signal amplification, further improve the sensitivity of HCV antibody test with a plurality of signal reporter molecules.
(3) in dual-antigen sandwich method, non-special absorption or with the human antibody of the non-specific bond of solid-phase reagent can be by the antigen recognizing of micromolecule mark, thereby improved the specificity of HCV antibody test significantly;
(4) this method specificity is good, thereby detection method involved in the present invention need not diluted sample, and sample is directly used in detection, helps detecting of HCV antigen/antibody combination;
Except the antibody that detects HCV IgG type, dual-antigen sandwich method also can detect the antibody of a plurality of types such as IgM, IgE, IgA; Especially IgM detection of antibodies helps shortening the window phase of HCV antibody test.
Four, embodiment:
The preparation of embodiment 1 biotin labeling HCV antigen
Step is as follows:
(1) with the HCV recombinant antigen in room temperature (+20～+ 25 ℃) to pH 7.4 100mM PBS (containing 3M urea) dialysis 24 hours, change liquid 2 times, each 2,000mL.Adjust concentration to about 1mg/mL.
(2) getting the Biotin-cap-NHS of aequum, be dissolved into 40mg/mL with DMF, is 1: 2 ratio in Biotin-cap-NHS and antigenic quality ratio, joins in the HCV recombinant antigen solution after the dialysis, fast mixing at once.Room temperature (+20～+ 25 ℃) left standstill 1 hour.
(3) room temperature (+20～+ 25 ℃) is to pH 7.8, and 50mM TSA (containing 3M urea) fully stirs dialysis 24 hours, changes liquid 2 times, and is each 1,000mL.Obtain the biotin labeling HCV antigen of purifying.
The preparation of embodiment 2 Eu mark streptavidins
Step is as follows:
(1) streptavidin 1mg is dissolved in 2,000 μ L pure water, room temperature (+20～+ 25 ℃) is changed liquid twice to pH 9.6 100mM CBS dialysis 24 hours, and is each 2,000mL.
(2) get 1mg Eu-DTTA, dissolve with 1000 μ L pH, 9.5 100mM CBS.With in the streptavidin solution that dissolves after Eu-DTTA adding is completely dialysed (about 1mg/mL), the vibration of limit edged, room temperature (+20～+ 25 ℃) left standstill 72 hours.The mass ratio of Eu-DTTA and streptavidin is about 1: 1.
(3) (1.6 * 50cm) separate Eu-SA and free Eu-DTTA, with pH 7.850mM TSA wash-out with Sephacryl S-200 HR.Detachment process is monitored with the nucleic acid-protein detector.Obtain the Eu mark streptavidin of purifying.
The preparation of the antigen coated microwell plate of embodiment 3 HCV
Step is as follows:
(1) pipette an amount of HCV recombinant antigen and be cushioned in the liquid in volume required bag, fully mixing avoids producing bubble, is mixed with the antigen coated liquid of HCV;
(2) get the every hole of microwell plate and add the antigen coated working fluid 100 μ L of HCV, room temperature (+20～+ 25 ℃) left standstill 24 hours;
(3) washing is 2 times; Sealing damping fluid 300 μ L, room temperature (+20～+ 25 ℃) standing over night (about 16 hours) are injected in every hole;
(4) blot the sealing damping fluid, every hole adds 4% (w/v) sucrose solution, 300 μ L, blots, and plank is put into drier dry 5 minutes; Open freeze dryer, make vacuum tightness＜50Pa, kept 3 hours, shut down, 2～8 ℃ of preservations are put in pack into tin platinum bag and the heat-sealing of dry afterreaction plate.
The total detection of antibodies of embodiment 4 HCV
Step is as follows:
(1) the corresponding micropore of sample is numbered according to the order of sequence, every plate should be established anti-HCV negative control, positive control (each 2 hole of or linear reference product A～F).
(2) sample or the calibration object of absorption 25 μ L are sequentially added into corresponding micropore.
(3) add 100 μ L biotin labeling antigen working fluids in each micropore, room temperature (+20～+ 25 ℃) was vibrated at a slow speed 45 minutes.
(4) preparation Eu mark streptavidin working fluid: to each bar 12 orifice plate, get 75 μ L Eu mark streptavidins (21 *), add the 1.5mL analysis buffer, fully mixing (dilution in 1: 21).Second Buwen educates preparation in preceding 45 minutes.
(5) wash plate 5 times, lath is patted dry on the thieving paper of cleaning.
(6) add 100 μ LEu mark streptavidin working fluids in each micropore, room temperature (+20～+ 25 ℃) was vibrated at a slow speed 15 minutes.
(7) wash plate 6 times, lath is patted dry on the thieving paper of cleaning.
(8) the enhancing liquid of adding 100 μ L in each micropore, room temperature (+20～+ 25 ℃) was vibrated at a slow speed 5 minutes.
(9) select corresponding program to survey fluorescence intensity.
Three batches based on the total antibody diagnosing reagent kit of double antigens sandwich HCV of the micromolecule biotin indirect labelling mensuration to national Panel (lot number 0401):
Specificity: 30 parts of negative reference products that Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides, 0 part of false positive, specificity is 100%, meets the requirements fully.
Sensitivity: 30 parts of positive reference products that Nat'l Pharmaceutical ﹠ Biological Products Control Institute provides, detect 29 parts, sensitivity is 96.7%, meets the requirements.4 parts of sensitivity reference materials, L1, L2 and L3 inspection are positive, and the L4 inspection is negative, meets the requirements fully.
Accuracy: 1 part of anti-HCV accuracy Quality Control product, replicate determination 10 holes, CV＜10% meets the requirements fully.
Reagent still can reach the sensitivity and the specificity of national regulation by 37 ℃ of bakings in 7 days.
Based on the total antibody diagnosing reagent kit of double antigens sandwich HCV of micromolecule biotin indirect labelling mensuration to Shanghai Blood Center Panel:
Sensitivity=15/ (15+0) * 100%=100%
Specificity=15/ (15+0) * 100%=100%
Coincidence rate=(15+15)/(15+0+15+0)=100%
Based on the total antibody diagnosing reagent kit of double antigens sandwich HCV of micromolecule biotin indirect labelling mensuration to the clinical examination center P anel of the Ministry of Public Health:
Sensitivity=24/ (24+1) * 100%=96%
Specificity=25/ (25+0) * 100%=100%
Coincidence rate=(24+25)/(24+0+25+0)=98%
Above result shows, based on the total antibody diagnosing reagent kit of HCV of micromolecule biotin indirect labelling in the requirement that all can reach clinical trial and blood screening aspect sensitivity and the specificity.
To the detection of 100 parts of confirmed anti-HCV Quality Control reference products of Chinese biological products assay institute, with the comparison of domestic anti-HCV ELISA diagnostic kit.
With the domestic commercially available K anti-HCV ELISA of company diagnostic kit (lot number: 20031205,20040602), the anti-HCV ELISA of W company diagnostic kit (lot number: C20040404 the effect phase:, 20041001) and the anti-HCV ELISA of X company diagnostic kit (lot number: SHJ020428) relatively the effect phase:, testing result to 100 parts of confirmed anti-HCV Quality Control reference products of Chinese biological products assay institute (45 parts of positives and 55 parts of feminine genders), see the following form:
Anti-HCV ELISA indirect method based on the total antibody diagnosing reagent kit of double antigens sandwich HCV of micromolecule biotin indirect labelling and K, W, X three companies detects result's contrast of calibrating institute anti-HCV Quality Control reference product:
As seen from the above table, all obviously be better than homemade anti-HCV ELISA diagnostic kit based on total antibody diagnosing reagent kit sensitivity of the double antigens sandwich HCV of micromolecule biotin indirect labelling and specificity, especially with the obvious advantage aspect specificity.
The total antibody diagnosing reagent kit of double antigens sandwich HCV and K, W, 15 parts of HCV-IgM antibody positives of anti-HCV ELISA indirect method detection of X three companies, HCV RNA positive sample result based on micromolecule biotin indirect labelling contrast:
As seen from the above table, for HCV-IgM antibody positive, HCV RNA positive sample, the recall rate based on the total antibody diagnosing reagent kit of double antigens sandwich HCV of micromolecule biotin indirect labelling among the present invention obviously is better than homemade ELISA reagent.
Can find out by above-mentioned experimental result:
(1) is that indirect labelling detects HCV antibody with the micromolecule, improved the sensitivity of HCV detection of antibodies effectively.
(2) be that indirect labelling detects HCV antibody with the micromolecule, improved HCV detection of antibodies specificity effectively.
(3) be the dual-antigen sandwich method detection HCV antibody of indirect labelling with the micromolecule, can not only detect the antibody of HCV IgG type, also help the antibody of a plurality of types such as detecting IgM, IgE, IgA.
Claims (9)
1. a micro-molecule indirectly labeled double antigens sandwich is measured the hepatitis C virus total antibody method, it is characterized in that method may further comprise the steps:
A) with small-molecule substance mark C hepatitis virus antigen;
B) with the C hepatitis virus antigen bag by solid phase material;
C) generate the anti-small molecular antibody of material mark or other materials that can combine with signal, be prepared into the signal reporter molecules with micromolecule;
D) utilize the immune response of antibody of HCV in the C hepatitis virus antigen of the C hepatitis virus antigen on solid phase material surface and micromolecule mark and the sample, form the double antigens sandwich compound: C hepatitis virus antigen 1-antibody of HCV-C hepatitis virus antigen 2-micromolecule; Utilize the association reaction of micromolecule and signal reporter molecules then, make the compound of formation have detectable signal generation material, be used for detecting the content of sample hepatitis C virus total antibody.
2. micro-molecule indirectly labeled double antigens sandwich according to claim 1 is measured the hepatitis C virus total antibody method, it is characterized in that the described small-molecule substance of step a) comprises:
(1) biotin molecule;
(2) digoxin, fluorescein, thyroxine, estradiol, estriol, triiodo thryonine, testosterone, progesterone.
3. micro-molecule indirectly labeled double antigens sandwich according to claim 1 is measured the hepatitis C virus total antibody method, it is characterized in that: step a), b) described C hepatitis virus antigen refers to gene recombinant antigens or synthetic peptide.
4. micro-molecule indirectly labeled double antigens sandwich according to claim 1 is measured the hepatitis C virus total antibody method, and it is characterized in that: the described solid phase material of step b) refers to micro reaction plate, test tube, plastic grain or plastic particles, magnetic particle.
5. micro-molecule indirectly labeled double antigens sandwich according to claim 1 is measured the hepatitis C virus total antibody method, it is characterized in that the described C hepatitis virus antigen bag of step b) by the step of solid phase material is:
(1) C hepatitis virus antigen is dissolved in bag and is cushioned liquid, be cushioned liquid with bag and soak solid phase material; Left standstill 12～24 hours;
(2) with the damping fluid washing solid phase material that contains surfactant; Add the damping fluid of casein containing protein, left standstill 3～12 hours;
(3) discard bag and be cushioned liquid, dry solid phase material is finished reorganization C hepatitis virus antigen bag by the preparation of solid phase material.
6. micro-molecule indirectly labeled double antigens sandwich according to claim 1 is measured the hepatitis C virus total antibody method, it is characterized in that: the described signal of step c) generates material and refers to enzyme, fluorescent material, rare earth ion such as Eu
3+, Sm
3+, Tb
3+, Dy
3+And chelate aglucon; The signal reporter molecules refers to that signal generates anti-small molecular antibody or other molecule that can combine with micromolecule of material mark.
7. micro-molecule indirectly labeled double antigens sandwich according to claim 1 is measured the hepatitis C virus total antibody method, it is characterized in that: the described hepatitis C virus total antibody of step d) refers to by various IgG, IgM that hepatitis C virus caused, IgE, the IgA antibody of invading human body.
8. micro-molecule indirectly labeled double antigens sandwich according to claim 1 is measured the hepatitis C virus total antibody method, it is characterized in that the step of hepatitis C virus total antibody in the described detection sample of step d) is:
(1) the direct sample that in container, adds not diluted;
(2) C hepatitis virus antigen of adding micromolecule mark; The C hepatitis virus antigen of the C hepatitis virus antigen on solid phase material surface and micromolecule mark and the reaction of the antibody of HCV in the sample;
(3) washing solid phase material; Add the signal reporter molecules, it is combined with the micromolecule label on solid phase material surface;
(4) washing solid phase material; Cause and measuring-signal.
9. micro-molecule indirectly labeled double antigens sandwich according to claim 1 is measured the hepatitis C virus total antibody method, and it is characterized in that: the described sample of step d) refers to whole blood, serum, blood plasma, urine, saliva.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200610027230A CN100582780C (en) | 2006-06-02 | 2006-06-02 | Micro-molecule indirectly labeled dual-antigen sandwich method for determining hepatitis C virus total antibody |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200610027230A CN100582780C (en) | 2006-06-02 | 2006-06-02 | Micro-molecule indirectly labeled dual-antigen sandwich method for determining hepatitis C virus total antibody |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1885039A CN1885039A (en) | 2006-12-27 |
| CN100582780C true CN100582780C (en) | 2010-01-20 |
Family
ID=37583303
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200610027230A Active CN100582780C (en) | 2006-06-02 | 2006-06-02 | Micro-molecule indirectly labeled dual-antigen sandwich method for determining hepatitis C virus total antibody |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100582780C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018133038A1 (en) | 2017-01-20 | 2018-07-26 | 深圳市新产业生物医学工程股份有限公司 | Labelled complex and preparation method therefor, and kit, use and detection system thereof |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101762694A (en) * | 2009-06-24 | 2010-06-30 | 北京科美东雅生物技术有限公司 | Magnetic immunochromatographic strip for detection of HCV antibody and preparation method thereof |
| CN101968483A (en) * | 2009-07-27 | 2011-02-09 | 深圳市菲鹏生物股份有限公司 | One-step semi-double-antigen sandwich immunological detection method |
| CN102236020A (en) * | 2010-04-20 | 2011-11-09 | 上海新波生物技术有限公司 | Hepatitis c virus antibody time-resolved fluoroimmunoassay and kit |
| CN102262158A (en) * | 2010-05-27 | 2011-11-30 | 戴国祯 | Direct application of recombinant fusion protein to quick diagnostic detection |
| CN103175963A (en) * | 2011-12-26 | 2013-06-26 | 苏州新波生物技术有限公司 | Treponema (TP) antibody detection method and detection kit thereof |
| CN102928595A (en) * | 2012-10-12 | 2013-02-13 | 武汉康苑生物医药科技有限公司 | Joint detection and detection kit for hepatitis c virus antigent antibody by using time-resolved fluoroimmunoassay |
| CN105486856A (en) * | 2015-12-24 | 2016-04-13 | 泰州泽成生物技术有限公司 | Qualitative detection kit for antibody to hepatitis C (anti-HCV) and preparing method and application thereof |
| CN105445451B (en) * | 2016-01-08 | 2017-04-12 | 广州市丰华生物工程有限公司 | Sample adding method suitable for time resolution fluorescence immunoassay method |
| CN109705197A (en) * | 2018-12-19 | 2019-05-03 | 广东菲鹏生物有限公司 | Hepatitis C Virus recombinant antigen and its application |
| CN110456073B (en) * | 2019-08-21 | 2023-09-22 | 广东菲鹏生物有限公司 | Method and kit for detecting antibody by double antigen sandwiches |
| CN113588940B (en) * | 2021-07-30 | 2023-05-12 | 菲鹏生物股份有限公司 | Method and kit for improving sensitivity of indirect labeling |
-
2006
- 2006-06-02 CN CN200610027230A patent/CN100582780C/en active Active
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018133038A1 (en) | 2017-01-20 | 2018-07-26 | 深圳市新产业生物医学工程股份有限公司 | Labelled complex and preparation method therefor, and kit, use and detection system thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1885039A (en) | 2006-12-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN100582780C (en) | Micro-molecule indirectly labeled dual-antigen sandwich method for determining hepatitis C virus total antibody | |
| CN104697988B (en) | Detect kit and its detection method and the application of antibody of HCV | |
| FI106317B (en) | Hepatitis C virus (HCV) antigen combinations for use in immunoassays of anti-HCV antibodies | |
| CN101196518B (en) | Hepatitis virus type C immune body chemiluminescence method diagnostic reagent kit and its producing method | |
| CN104698172B (en) | Kit for detecting hepatitis B surface antigen | |
| CN103175963A (en) | Treponema (TP) antibody detection method and detection kit thereof | |
| CN101713780A (en) | Method for magnetic antibody immunoassay chemiluminescence detection of treponema pallidum antibodies | |
| CN104090105A (en) | Method and kit for detecting hepatitis E virus (HEV) antibody and method for preparing kit | |
| CN102236020A (en) | Hepatitis c virus antibody time-resolved fluoroimmunoassay and kit | |
| CN103293299A (en) | Method and kit for broadening double-antibody sandwich immunodetection concentration range | |
| CN104198721B (en) | The preparation of a kind of silica-based magnetic bead bond of GP73 (GP73) antigen and application | |
| CN105445463A (en) | Hepatitis E virus IgG antibody detection kit and preparation method and application thereof | |
| CN101750502A (en) | TRFIA for synchronously detecting AFP and AFP-IgM and reagent kit thereof | |
| JP2018501483A (en) | Method for reducing interference | |
| CN101498730B (en) | Improved double-antigen sandwiching immunity detection method | |
| CN101382553A (en) | Large protein pre-S surface antigen for hepatitis B virus chemiluminescence immune assay kit and method for making same | |
| CN103048456A (en) | Hepatitis B virus PreS1 antigen enzyme-linked immunoassay kit employing one-step method | |
| CN102654505A (en) | Time-resolved fluorescence immunoassay reagent kit used for detecting interleukin (IL)-2-human serum albumin (HSA) and detection method thereof | |
| CN102095846A (en) | Chemiluminescence quantitative detection kit for carbohydrate antigen 242 | |
| CN101368962A (en) | Chemical luminescence immune assay determination reagent kit for prostate gland acid phosphatase and preparation method thereof | |
| CN101368964A (en) | Chemical luminescence method immune analysis diagnostic reagent kit for detecting cytomegalovirus IgG antibody | |
| CN101368965A (en) | Chemical luminescence method immune analysis diagnostic reagent kit for detecting cytomegalovirus IgM antibody | |
| CN106119340A (en) | Detection method, detectable and the detection kit of a kind of de-γ carboxyl thrombinogen | |
| CN102590505A (en) | Reagent kit for detecting immunoglobulin m (IgM) AlphaLISA for resisting enterovirus71 capsid protein1 | |
| CN114720700A (en) | Application of reagent for detecting anti-cytoskeleton-associated protein4-IgG autoantibody in preparation of kit for detecting vascular endothelial injury |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| ASS | Succession or assignment of patent right |
Free format text: FORMER OWNER: SUZHOU SYM-BIO LIFESCIENCE CO., LTD. Effective date: 20120215 Owner name: SUZHOU SYM-BIO LIFESCIENCE CO., LTD. Free format text: FORMER OWNER: SHANGHAI XINBO BIOTECHNOLOGY CO., LTD. Effective date: 20120215 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| COR | Change of bibliographic data |
Free format text: CORRECT: ADDRESS; FROM: 201203 PUDONG NEW AREA, SHANGHAI TO: 215400 SUZHOU, JIANGSU PROVINCE |
|
| TR01 | Transfer of patent right |
Effective date of registration: 20120215 Address after: 215400 No. 36, Luoyang Road, Taicang, Jiangsu Patentee after: Suzhou Sym-Bio Lifescience Co., Ltd. Address before: 201203 Shanghai City, Pudong New Area Zhangjiang Road No. 351, No. 2 GuoShouJing Building Room 502 Co-patentee before: Suzhou Sym-Bio Lifescience Co., Ltd. Patentee before: Shanghai Xinbo Biotechnology Co., Ltd. |