CN201477095U - Sensitive and fast cirrhosis and liver cancer magnetic bead enzyme-free diagnostic reagent kit and magnetic beads - Google Patents
Sensitive and fast cirrhosis and liver cancer magnetic bead enzyme-free diagnostic reagent kit and magnetic beads Download PDFInfo
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- CN201477095U CN201477095U CN2009202223403U CN200920222340U CN201477095U CN 201477095 U CN201477095 U CN 201477095U CN 2009202223403 U CN2009202223403 U CN 2009202223403U CN 200920222340 U CN200920222340 U CN 200920222340U CN 201477095 U CN201477095 U CN 201477095U
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- magnetic bead
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- liver cancer
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Abstract
The utility model relates to a sensitive and fast cirrhosis and liver cancer magnetic bead enzyme-free diagnostic reagent kit and magnetic beads, which belongs to the filed of medical diagnosis. The sensitive and fast cirrhosis and liver cancer magnetic bead enzyme-free diagnostic reagent kit comprises a kit body and a reagent pipe bracket, wherein the reagent pipe bracket is pasted on the inner wall of the kit body, the upper surface of the reagent pipe bracket is provided with a plurality of reagent pipe grooves, a magnetic bead pipe provided with magnetic beads is placed in one reagent pipe groove, and an elisa plate is placed in the other reagent pipe groove. The utility model is characterized in that a layer of antibody is covered on the surfaces of the magnetic beads, the elisa plate is a blank micropore plate which is used for the reaction of the magnetic beads and the blood serum to be tested in plate holes of the blank micropore plate, the antibody refers to the antibody of indicative antigen protein HCCR-2 special combination in the blood serum of cirrhosis patients, or refers to the antibody of indicative antigen protein HCCR-1 special combination in the blood serum of liver cancer patients.
Description
Technical field
The utility model relates to the medical diagnosis reagent box, and particularly a kind of sensitivity cirrhosis and liver cancer magnetic bead enzyme is fast exempted from method diagnostic kit and magnetic bead
Background technology
All types of hepatites virus infections, excessively take in alcohol and be in for a long time in the environment that is rich in organic solvent and can cause hepatic injury, the result of hepatic injury is the formation of inflammation, the hardening of tissue or even cancer.Liver cancer mortality occupies the 3rd after being listed in lung cancer, cancer of the stomach in malignant tumour.Liver cancer serious harm China people's life security, therefore, be necessary very much deep research is done in the early diagnosis of cirrhosis and liver cancer.
In the diagnostic techniques commonly used at present, the false positive height, detection sensitivity is not enough, the capital is brought to the patient and is seriously influenced, development along with Enzyme-multiplied immune technique, it is found that specifically binding to Ag-Ab association reaction that the antibody from people's proto-oncogene HCCR-1 and HCCR-2 expressed proteins carries out by use can distinguish and effectively detect early liver cancer and cirrhosis, (Di Dongjin, people such as Chen Junhao, HCCR1 and HCCR2 soluble protein are expressed in bacterium, purifying and the application in the diagnosing cancer of liver in early days, the contemporary Chinese medical journal, 2006 the 16th the 10th phases of volume), in Chinese patent application numbers 200480014598.4, the Chinese patent application of " diagnostic kit for liver cirrhosis that comprises specific antibody at people's proto-oncogene protein " provides a kind of traditional ELISA Plate enzyme linked immunological absorption assay method (ELISA) to detect the method for HCCR-2 antigen in the sample, and (YoonSK in 2006, Lim NK, et The Human Cervical Cancer Oncogene Protein Is a Biomarker for HumanHepatocellular Carcinoma.Cancer Research 64,5434-5441, August 1,2004), also adopt traditional ELISA Plate enzyme linked immunological absorption assay method (ELISA) to detect HCCR-1 antigen in the sample, but traditional board-like ELISA detection method has sensitivity not high, operate shortcomings such as length consuming time, these two antigen proteins are that early detection cirrhosis and liver cancer provide perfect mark, but present detection method has limited its detection sensitivity and accuracy.There are some researches show that on the particulate magnetic bead dressing probe or antibody carry out the experiment of purposes such as separation and purification, having can be quantitative, easy to operate and disturbing factor is few, advantages such as sensitivity and specificity are good, but do not wrap by the magnetic bead of the antibody of cirrhosis and liver cancer marker protein at present, do not comprise the kit that is suitable for detecting cirrhosis and liver cancer of similar magnetic bead yet.
The utility model content
Blank according to above-mentioned field, the utility model provides sensitivity, and cirrhosis and liver cancer magnetic bead enzyme are exempted from method diagnostic kit and magnetic bead fast, the magnetic bead bag is by the antibody of significant antigen protein in anti-cirrhosis and the liver cancer patient blood serum, this magnetic bead and other detectable are placed in the box with specific locus, the design of science makes and is positioned over wherein each kind of reagent and in use can stablizes and preserve actively, guarantees the sensitivity and the accuracy of magnetic bead detection reaction.
Sensitivity cirrhosis and liver cancer magnetic bead enzyme is fast exempted from method diagnostic kit and magnetic bead, comprise box body and the reagent pipe holder of fitting with inboard wall of cartridge, described reagent pipe holder upper surface is provided with some reagent tube seats, be placed with the magnetic bead pipe that fills magnetic bead in described one of them reagent tube seat, place ELISA Plate in another reagent tube seat, it is characterized in that, described magnetic bead surfaces is coated with one deck antibody, described ELISA Plate is the blank microwell plate that magnetic bead and test serum react in its plate hole, described antibody refers to the antibody of significant antigen protein HCCR-2 specific bond in the liver cirrhosis patient serum, or refers to the antibody of significant antigen HCCR-1 specific bond in the liver cancer patient blood serum.
The reagent tube seat that is provided with on the described reagent pipe holder refers to the chromogenic substrate tube seat, label tube seat, damping fluid tube seat, reaction terminating liquid tube seat, contrast liquid tube seat and/or magnetic force frame groove.
Be placed with the chromogenic substrate pipe in the chromogenic substrate tube seat on the described reagent pipe holder, be placed with the mark property management in the label tube seat, be placed with the damping fluid pipe in the damping fluid tube seat, be placed with reaction stop solution pipe in the reaction stop solution tube seat, be placed with in contrast liquid pipe and/or the magnetic force frame groove in the contrast liquid tube seat and be placed with the magnetic force frame.Described ELISA Plate groove and magnetic bead tube seat region are positioned at the same end of reagent pipe holder, described zone be one with the removable carriage that is connected of reagent pipe holder other parts.
A kind of magnetic bead is characterized in that: described magnetic bead surfaces is coated with the antibody that one deck has the antigen recognizing activity.
The utility model sensitivity cirrhosis and liver cancer magnetic bead enzyme is fast exempted from the method diagnostic kit, for early detection cirrhosis and liver cancer provide a kind of convenient, efficient and high approach of detection sensitivity, the interior ELISA Plate of this kit is coated antibody not, but wrap by bead particulates with antibody, the bead particulates of bag quilt is used as reagent, is more conducive to quantitative work, and is convenient stable, and sensitivity is higher, and disturbing factor can drop to minimum in the antibody antigen reaction.Comprise the magnetic bead pipe and the ELISA Plate that fill magnetic bead in the kit, magnetic bead pipe and ELISA Plate are installed on the interior reagent pipe holder of box, because magnetic bead pipe and ELISA Plate are to use simultaneously in detection, indispensable, and diversity ratio is bigger in shape between the two, if at will be placed in the box, clash into mutually easily when mobile and in the transportation and cause magnetic bead character instability, and then influence the reliability of testing result, therefore the utility model is provided with a reagent pipe holder in box, and the reagent pipe holder is provided with and the stable groove that engages of reagent pipe and ELISA Plate bottom.Can guarantee that like this magnetic bead that is coated with antibody in the utility model kit keeps stable activity, thereby guarantee the sensitivity of antibody antigen reaction.
Can also be provided with on the reagent pipe holder in the utility model kit and need all ingredients used and the groove of equipment in the detection reaction, be convenient in kit, put into all ingredients and the equipment that to use, as chromogenic substrate, label, damping fluid, reaction stop solution, contrast liquid, magnetic force frame etc.It is in use that these grooves are set, and makes things convenient for the experimenter to dispose all ingredients as required and is placed in the kit, easy to use and carry.
The utility model is the required reagent corresponding pipe of configuration detection reaction in the tube seat of each in kit preferably, improves the applicability and the convenient degree of this kit.
Preferred described ELISA Plate groove of the utility model and magnetic bead tube seat region are positioned at the same end of reagent pipe holder, and described zone is removable the connection with reagent pipe holder other parts.Owing in detection reaction, use magnetic bead and ELISA Plate at first, and during 20～30 minutes of antibody antigen reaction, other reagent in the kit all need not, if whole kit is taken out from refrigerator, destroy the stability of reagent easily because of the reagent multigelation, therefore, magnetic bead tube seat and ELISA Plate groove is independent of a zone, described zone make with the reagent pipe holder between a removable carriage that is connected, when carrying out the first step of detection reaction, only take out carriage, and do not influence other reagent in the box.
Description of drawings:
The 1-box body, 2-reagent pipe holder, 3-magnetic bead pipe, 3A-magnetic bead tube seat, 4-ELISA Plate, 4A-ELISA Plate groove, 5-reagent tube seat, 5A-reagent pipe, 6-chromogenic substrate tube seat, 7-label tube seat, 8-damping fluid tube seat, 9-reaction stop solution tube seat, 10-contrasts liquid, 11-magnetic force frame groove, 12-carriage.
Fig. 1. the utility model kit is opened the schematic top plan view behind the lid.
Fig. 2. the schematic top plan view after the utility model kit is uncapped behind the reject reagent pipe wherein.
Fig. 3. splendid attire ELISA Plate and magnetic bead pipe and the schematic top plan view removable join domain of reagent pipe holder.
Fig. 4. the structural representation of magnetic bead coated antibody in the utility model kit.
Embodiment:
Embodiment 1.
As shown in Figures 1 and 2, the utility model sensitivity cirrhosis and liver cancer magnetic bead enzyme is fast exempted from the method diagnostic kit, comprise box body 1 and the reagent pipe holder 2 of fitting with inboard wall of cartridge, described reagent pipe holder 2 upper surfaces are provided with some reagent tube seats 5, described one of them reagent tube seat is to be placed with the magnetic bead pipe 3 that fills magnetic bead among the magnetic bead tube seat 3A, place ELISA Plate 4 among another reagent tube seat and the ELISA Plate groove 4A, it is characterized in that, it is characterized in that, described magnetic bead surfaces is coated with one deck antibody, described ELISA Plate 4 is the blank microwell plate that magnetic bead and test serum react in its plate hole, described antibody refers to the antibody of significant antigen protein HCCR-2 specific bond in the liver cirrhosis patient serum, or refers to the antibody of significant antigen HCCR-1 specific bond in the liver cancer patient blood serum.Comprise the magnetic bead pipe 3 and the ELISA Plate 4 that fill magnetic bead in the utility model kit, magnetic bead pipe 3 and ELISA Plate 4 are installed on the interior reagent pipe holder 2 of box, because magnetic bead pipe 3 uses simultaneously with ELISA Plate 4 in detection, indispensable, and diversity ratio is bigger in shape between the two, if at will be placed in the box, clash into mutually easily when mobile and in the transportation and cause magnetic bead character instability, and then influence the reliability of testing result, therefore the utility model is provided with a reagent pipe holder 2 in box, and reagent pipe holder 2 is provided with and the stable groove that engages in the bottom of magnetic bead pipe 3 and ELISA Plate 4.Can guarantee that like this magnetic bead that is coated with antibody in the utility model kit keeps stable activity, thereby guarantee antibody-antigen reactive sensitivity, be provided with and the stable groove that engages in the bottom of magnetic bead pipe 3 and ELISA Plate 4.Can guarantee that like this magnetic bead that is coated with antibody in the utility model kit keeps stable activity, thereby guarantee antibody-antigen reactive sensitivity, bag is by the antibody of antigen protein of anti-sign cirrhosis and liver cancer on the magnetic bead, being used as reagent in the antibody antigen reaction uses, be convenient to quantitative and detection, it can discern distinctive sign antigen protein in cirrhosis or the liver cancer patient blood serum, thereby realizes the early detection of cirrhosis and liver cancer.
The antibody of the magnetic bead bag quilt in the magnetic bead pipe in the utility model kit is the monoclonal antibody of the antigen protein HCCR-2 albumen of anti-sign cirrhosis, or the monoclonal antibody of the antigen protein HCCR-1 of anti-sign liver cancer.
The utility model is in order to provide kit more easily for the testing personnel, in this kit, the reagent tube seat 5 that is provided with on the reagent pipe holder 2 also comprises chromogenic substrate tube seat 6, label tube seat 7, damping fluid tube seat 8, enzymatic reaction stop solution tube seat 9, contrast liquid tube seat 10 and/or magnetic force frame groove 11.In use, the experimenter can be as required oneself configuration all ingredients be placed in the kit, easy to use and carry.
Further, kit of the present utility model, be placed on the chromogenic substrate pipe in the chromogenic substrate tube seat 6 on the described reagent pipe holder in addition, mark property management in the label tube seat 7, damping fluid pipe in the damping fluid tube seat 8, reaction stop solution pipe in the reaction stop solution tube seat 9, the contrast liquid pipe in the contrast liquid tube seat 10 and/or be positioned over magnetic force frame in the magnetic force frame groove 11.Directly provide testing cost required detectable for the testing personnel.
Further decision design such as Fig. 2, shown in Figure 3, described ELISA Plate groove 4A and magnetic bead tube seat 3A region are positioned at the same end of reagent pipe holder 2, and it is the removable carriage that is connected 12 with the other parts of reagent pipe holder 2 that described zone is one.Owing in detection reaction, use magnetic bead pipe 3 and ELISA Plate 4 at first, and during 20～30 minutes of antibody antigen reaction, other reagent in the kit all need not, if whole kit is taken out from refrigerator, destroy easily the stability of reagent, when carrying out the first step antigen-antibody binding reaction of detection reaction, only take out carriage 12 and get final product because of reagent multigelation or temperature variation, and need not this box be taken out, therefore do not influence other reagent in the box.
2.1 sample requires: human serum, require limpid, do not have precipitation, a no jaundice; 1 week of sample can store under 4 ℃ of conditions with interior detection; 1 week of sample, above the detection need be frozen under-20 ℃ of conditions, and avoid multigelation.
2.2 from kit, take out carriage 6, and the magnetic bead pipe on the taking-up carriage 3 is a microwell plate with ELISA Plate 4, join the magnetic bead that is coated with the HCCR-2 monoclonal antibody in the magnetic bead pipe 3 such as Fig. 4 in the microwell plate rapidly, each hole adds 90ul, and adding test serum 10ul respectively, 2-8 ℃ of concussion hatched 30 minutes.
2.3 washing: microwell plate is placed on magnetic sheet last 2 minute to magnetic bead is adsorbed in tube wall, sop up supernatant.Use the PBS buffer solution for cleaning magnetic bead three times in the damping fluid pipe, operation is with common biotin labeled magnetic bead, used the resuspended magnetic bead of 100ul buffer A (0.1M Na-phosphate buffer pH 7.4) in the damping fluid pipe.
2.4 open the mark property management, draw label HRP (two is anti-), add 100ul HRP mark HCCR-2 monoclonal antibody in each micropore, reacted 15 minutes at 37 ℃ of incubators or water-bath concussion.
2.5 repeat 2.3 washings.
2.6 colour developing: take out the substrate solution that the sucking-off of substrate solution pipe can make the colour developing of previous step label, every hole adds substrate solution 50ul, keeps concussion, 10 minutes (adding fashionable picking up counting from first hole) of room temperature lucifuge accurate response.
2.7 stop: take out the reaction terminating liquid pipe, every reacting hole adds the 50ul stop buffer on the microwell plate.Slight concussion mixing.
2.8 microwell plate is put on the magnetic sheet, magnetic bead is inhaled in the lath sidewall.
2.9 value of reading: take off microwell plate gently, microplate reader is set in dual wavelength 450nm/630nm, measure each hole A value, value of reading in 15 minutes after cessation reaction.
Claims (5)
1. cirrhosis and liver cancer magnetic bead enzyme are exempted from the method diagnostic kit to sensitivity fast, comprise box body and the reagent pipe holder of fitting with inboard wall of cartridge, described reagent pipe holder upper surface is provided with some reagent tube seats, be placed with the magnetic bead pipe that fills magnetic bead in described one of them reagent tube seat, place ELISA Plate in another reagent tube seat, it is characterized in that, described magnetic bead surfaces is coated with one deck antibody, described ELISA Plate is the blank microwell plate that magnetic bead and test serum react in its plate hole, described antibody refers to the antibody of significant antigen protein HCCR-2 specific bond in the liver cirrhosis patient serum, or refers to the antibody of significant antigen HCCR-1 specific bond in the liver cancer patient blood serum.
2. kit according to claim 1 is characterized in that the reagent tube seat that is provided with on the described reagent pipe holder refers to the chromogenic substrate tube seat, label tube seat, damping fluid tube seat, reaction terminating liquid tube seat, contrast liquid tube seat and/or magnetic force frame groove.
3. kit according to claim 2, it is characterized in that being placed with the chromogenic substrate pipe in the chromogenic substrate tube seat on the described reagent pipe holder, be placed with the mark property management in the label tube seat, be placed with the damping fluid pipe in the damping fluid tube seat, be placed with reaction stop solution pipe in the reaction stop solution tube seat, be placed with in contrast liquid pipe and/or the magnetic force frame groove in the contrast liquid tube seat and be placed with the magnetic force frame.
4. according to claim 1 or 2 or 3 described kits, it is characterized in that described ELISA Plate groove and magnetic bead tube seat region are positioned at the same end of reagent pipe holder, described zone be one with the removable carriage that is connected of reagent pipe holder other parts.
5. magnetic bead, it is characterized in that: described magnetic bead surfaces is coated with the antibody that one deck has the antigen recognizing activity, described antibody refers to the antibody of significant antigen protein HCCR-2 specific bond in the liver cirrhosis patient serum, or refers to the antibody of significant antigen HCCR-1 specific bond in the liver cancer patient blood serum.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009202223403U CN201477095U (en) | 2009-09-02 | 2009-09-02 | Sensitive and fast cirrhosis and liver cancer magnetic bead enzyme-free diagnostic reagent kit and magnetic beads |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009202223403U CN201477095U (en) | 2009-09-02 | 2009-09-02 | Sensitive and fast cirrhosis and liver cancer magnetic bead enzyme-free diagnostic reagent kit and magnetic beads |
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| Publication Number | Publication Date |
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| CN201477095U true CN201477095U (en) | 2010-05-19 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009202223403U Expired - Lifetime CN201477095U (en) | 2009-09-02 | 2009-09-02 | Sensitive and fast cirrhosis and liver cancer magnetic bead enzyme-free diagnostic reagent kit and magnetic beads |
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| CN (1) | CN201477095U (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012028104A1 (en) * | 2010-09-02 | 2012-03-08 | 无锡人寰生物医药科技有限公司 | Single-dose intravenous injection kit of adult allogeneic bone marrow mesenchymal cell |
| CN102401831A (en) * | 2011-11-14 | 2012-04-04 | 北京正旦国际科技有限责任公司 | Competitive enzyme-linked immunosorbent assay kit for liver cirrhosis and assay method for competitive enzyme-linked immunosorbent assay kit |
| CN102520175A (en) * | 2011-11-14 | 2012-06-27 | 北京正旦国际科技有限责任公司 | Competitive enzyme-linked immunosorbent assay (ELISA) kit for liver cancer polypeptide marker antigen |
| CN103267851A (en) * | 2013-05-08 | 2013-08-28 | 成都创宜生物科技有限公司 | Kit for detecting premature rupture of membrane, and preparation method thereof |
| CN103293300A (en) * | 2012-07-09 | 2013-09-11 | 深圳市艾瑞生物科技有限公司 | Time resolution fluorescence detection kit based on phosphorescence luminous technology, and preparation method and application of detection kit |
| CN109283331A (en) * | 2017-11-15 | 2019-01-29 | 广州弗赛生物科技有限公司 | A method of the quick detection target based on colorimetric method and magnetic bead antibody beneficiation technologies |
-
2009
- 2009-09-02 CN CN2009202223403U patent/CN201477095U/en not_active Expired - Lifetime
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012028104A1 (en) * | 2010-09-02 | 2012-03-08 | 无锡人寰生物医药科技有限公司 | Single-dose intravenous injection kit of adult allogeneic bone marrow mesenchymal cell |
| CN102401831A (en) * | 2011-11-14 | 2012-04-04 | 北京正旦国际科技有限责任公司 | Competitive enzyme-linked immunosorbent assay kit for liver cirrhosis and assay method for competitive enzyme-linked immunosorbent assay kit |
| CN102520175A (en) * | 2011-11-14 | 2012-06-27 | 北京正旦国际科技有限责任公司 | Competitive enzyme-linked immunosorbent assay (ELISA) kit for liver cancer polypeptide marker antigen |
| CN102401831B (en) * | 2011-11-14 | 2013-10-30 | 北京正旦国际科技有限责任公司 | Competitive enzyme-linked immunosorbent assay kit for liver cirrhosis and assay method therefor |
| CN102520175B (en) * | 2011-11-14 | 2014-04-16 | 北京正旦国际科技有限责任公司 | Competitive enzyme-linked immunosorbent assay (ELISA) kit for liver cancer polypeptide marker antigen |
| CN103293300A (en) * | 2012-07-09 | 2013-09-11 | 深圳市艾瑞生物科技有限公司 | Time resolution fluorescence detection kit based on phosphorescence luminous technology, and preparation method and application of detection kit |
| CN103267851A (en) * | 2013-05-08 | 2013-08-28 | 成都创宜生物科技有限公司 | Kit for detecting premature rupture of membrane, and preparation method thereof |
| CN109283331A (en) * | 2017-11-15 | 2019-01-29 | 广州弗赛生物科技有限公司 | A method of the quick detection target based on colorimetric method and magnetic bead antibody beneficiation technologies |
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CX01 | Expiry of patent term |
Granted publication date: 20100519 |
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| CX01 | Expiry of patent term |