CN1166953C - Fast gold mark test reagent kit for one-step multiple-index test of myocardial infarction - Google Patents
Fast gold mark test reagent kit for one-step multiple-index test of myocardial infarction Download PDFInfo
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- CN1166953C CN1166953C CNB011130237A CN01113023A CN1166953C CN 1166953 C CN1166953 C CN 1166953C CN B011130237 A CNB011130237 A CN B011130237A CN 01113023 A CN01113023 A CN 01113023A CN 1166953 C CN1166953 C CN 1166953C
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- 239000010931 gold Substances 0.000 title claims abstract description 33
- 229910052737 gold Inorganic materials 0.000 title claims abstract description 33
- 239000003153 chemical reaction reagent Substances 0.000 title abstract description 9
- 208000010125 myocardial infarction Diseases 0.000 title abstract 4
- 238000012360 testing method Methods 0.000 title description 11
- 238000001514 detection method Methods 0.000 claims abstract description 35
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims abstract description 22
- 206010061216 Infarction Diseases 0.000 claims abstract description 17
- 230000007574 infarction Effects 0.000 claims abstract description 17
- 229960002685 biotin Drugs 0.000 claims abstract description 11
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- 239000011616 biotin Substances 0.000 claims abstract description 11
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- 102000004420 Creatine Kinase Human genes 0.000 description 1
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- LRBQNJMCXXYXIU-PPKXGCFTSA-N Penta-digallate-beta-D-glucose Natural products OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-PPKXGCFTSA-N 0.000 description 1
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- IZEKFCXSFNUWAM-UHFFFAOYSA-N dipyridamole Chemical compound C=12N=C(N(CCO)CCO)N=C(N3CCCCC3)C2=NC(N(CCO)CCO)=NC=1N1CCCCC1 IZEKFCXSFNUWAM-UHFFFAOYSA-N 0.000 description 1
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- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- FDWREHZXQUYJFJ-UHFFFAOYSA-M gold monochloride Chemical compound [Cl-].[Au+] FDWREHZXQUYJFJ-UHFFFAOYSA-M 0.000 description 1
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- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention relates to a reagent kit for rapidly detecting gold marks of miocardial infarction by multiple targets in a one step method, which comprises a bottom plate, a sample pad, water absorption paper and a fiber glass film fixed with gold mark antibodies, wherein an antibody II strip resisting A, a streptavidin strip and a strip resisting an antibody are respectively sprayed in a region on the bottom plate and between the fiber glass film and the water absorption paper at the upper end; a gold mark antibody coated by the fiber glass film contains a gold mark antibody I resisting A and a gold mark antibody I resisting B; an antibody II resisting B is also coated on the fiber glass film, and the antibody II resisting B is connected with biotin. The present invention combines two or more independent immune reaction systems for detecting markers of myocardial infarction into one detecting reagent kit of myocardial infarction. More than two project results for detecting myocardial infarction can be simultaneously obtained by one detection. The present invention has the advantages of detection sensitivity, accuracy and reliability.
Description
The present invention relates to biomedical sector, particularly a kind of one-step multiple-index mark of using in medical test detects the quick gold mark detection kit of miocardial infarction.
Immune colloidal gold technique is as a kind of new immunological method, and its development is very rapid, in biomedical each research field, has particularly obtained increasingly extensive application in medical test.Collaurum has become after fluorescein, emitting isotope and enzyme, and a kind of on-radiation comparatively commonly used in immunolabelling technique is marked agent.
Collaurum is the gold grain that is formed specific size by gold chloride polymerization under the effect of reductive agent such as tannic acid, trisodium citrate, and because electrostatic interaction becomes a kind of stable colloidal state, the characteristics that it has high electron density, can combine with multiple biomacromolecule.The immuno-gold labeling technology has mainly been utilized this feature of gold grain, be in the visible pitchy particle of microscopically in gold mark protein combination, when these mark things are assembled in a large number at corresponding part place, naked eyes red color visible or aubergine spot, thereby be used for qualitative or semiquantitative tachysynthesis detection method.
Since the gold-immunochromatographyreagent reagent for assay box with put exempt from, the enzyme marking reagent box compares, have and manipulate safe, easy, easy single part the detection and advantage such as quick, therefore the positive increased popularity of gold-labeled diagnosis reagent box in recent years, on some project, substituted gradually put exempt from, the enzyme marking reagent box, as very early pregnancy reagent strip (HCG Test Strip) etc.
Acute myocardial infarction (Acute Myocardial Infarction, AMI) state of an illness is dangerous, and the mortality ratio height is a kind of comparatively dangerous disease, and the major reason that AMI causes death is exactly can not early diagnosis, has incured loss through delay treatment opportunity.So it is in recent years, very active for the research of AMI mark.The biochemical marker of AMI includes myoglobins (Myoglobin, Mb), serum cardiac troponin T and cardiac muscle troponin I (cardiac Troponin T, cTnT, cardiac Troponin I, cTnI), creatine kinase (Myoglobin, MB) isodynamic enzyme (Cytokinase-Myoglobin, CK-MB), glycogen phosphorylase isoenzyme (GPBB) etc.
The cTnT gold mark detector bar that existing in the market like product has Roche Holding Ag (Roche) to produce, the serum Mb gold-marking immunity spotting method that the blue ripple biotechnology research in Sanming City, Fujian is developed etc.These products have following shortcoming:
1, certain heart that can only detect separately in the blood obstructs index.
2, the sensitivity of single project testing result and specificity are inferior slightly.
3 if obtain two or multinomial result and then need operate more than twice, comparatively trouble.
4, the production cost of two or more independent kits will be higher than a composite reagent kit.
5, the composition of these kits all is applicable to hospital diagnosis, and is not suitable for family, because the interior no blood taking needle of kit package, little suction pipe and vacuum-packed alcohol silk tops.
The purpose of this invention is to provide the quick gold mark detection kit that a kind of one-step multiple-index mark detects miocardial infarction, can obtain the test item result of two above miocardial infarctions by one-time detection simultaneously.
Technical scheme of the present invention is as follows:
A kind of one-step multiple-index mark detects the quick gold mark detection kit of miocardial infarction, it comprises base plate, the sample pad that is provided with in the base plate lower end, the thieving paper that is provided with in the base plate upper end, and at the glass fibre membrane of locating to post above the sample pad that is fixed with golden labeling antibody that is positioned at of base plate, be sprayed with anti-A antibody II band and antiantibody band respectively in the zone between glass fibre membrane and upper end thieving paper of described base plate, also be sprayed with the Streptavidin band in the zone between glass fibre membrane and upper end thieving paper of described base plate; The golden labeling antibody of described glass fibre membrane bag quilt comprises gold mark anti-A antibody I and gold mark anti-B antibody I, gold mark anti-A antibody I is anti-myoglobins antibody α, gold mark anti-B antibody I is anti-cardiac troponin antibody a, glass fibre membrane also wraps the anti-B antibody II that has been coupled biotin, and anti-B antibody II is anti-cardiac troponin antibody b; This kit adopts the separation of whole blood system, and after dripping whole blood on the sample pad, by the separation of separation of whole blood film, serum is separated to come, and further moves to the collaurum pad.
The composition of this kit also comprises check-out console, drying agent, blood taking needle, little suction pipe and vacuum inner wrapping sterilized cotton ball.
Described anti-A antibody II band bag is by myoglobins antibody β.
Described antiantibody band bag by sheep anti mouse two anti-.The present invention is combined to the two or more independent immune response system of thought-read muscle infarction mark in the detection kit of miocardial infarction, apply to gold mark check-out console or detector bar, the recall rate of its detection, sensitivity index meet the clinical detection requirement, have improved the accuracy and the reliability that detect.
Use when of the present invention, two or more miocardial infarction test items can be placed on the check-out console, one-time detection can obtain two or more test item results simultaneously, and is quick and easy, improved efficient.By the joint-detection of a plurality of projects, increased the specificity that detects, improved the sensitivity that detects, accurately, reliably, more help to make diagnosis accurately.
Product of the present invention is owing to having save the material of repetitive operation, so its production cost is lower than the cost of each individual event check-out console sum.
This product adopt to be fit to home-use packing, promptly adds blood taking needle in each kit, gets blood suction pipe, sterilized cotton ball, and patient is as feeling uncomfortable, and the detection of can taking a blood sample at home promptly went out the result in 10 minutes.This product, in time goes to hospital further to make a definite diagnosis and treat according to testing result and has positive help especially to the anginal patient of recessiveness the patient.Simultaneously, but patient's self-observation at home infraction once again whether has solved patient if there is discomfort must go hospital's queuing to do the problem of detection, has saved patient's the time to diagnose the illness.
Below in conjunction with drawings and Examples the present invention is elaborated.
Fig. 1 is the structural representation that a kind of one-step multiple-index mark detects the quick gold mark detection kit of miocardial infarction.
Fig. 2 is the schematic appearance of Fig. 1 detection kit.
Fig. 3 is the schematic diagram of Fig. 1 detection kit.
The testing result lexical or textual analysis of Fig. 4 presentation graphs 1 detection kit.
Referring to Fig. 1, the quick gold mark detection kit that a kind of one-step multiple-index mark detects miocardial infarction comprises base plate 1, the sample pad 2 that is provided with in base plate 1 lower end, the thieving paper 3 that is provided with in base plate 1 upper end, and base plate 1 be positioned at the glass fibre membrane that is fixed with golden labeling antibody 4 that place, sample pad 2 tops posts.
In the zone between glass fibre membrane 4 and upper end thieving paper 3 of base plate 1, be sprayed with anti-A antibody II band 5, Streptavidin band 6 and antiantibody band 7 respectively.
The golden labeling antibody of glass fibre membrane 4 bag quilts comprises gold mark anti-A antibody I and gold mark anti-B antibody I.Glass fibre membrane 4 also wraps the anti-B antibody II that has been coupled biotin.
In the present embodiment, gold mark anti-A antibody I is anti-myoglobins (Mb) antibody α.Gold mark anti-B antibody I is anti-cardiac troponin (cTnTb) antibody a.The anti-B antibody II that has connect biotin is anti-cardiac troponin (cTnTb) the antibody b that has connect biotin.Anti-A antibody II band 5 bags are by myoglobins (Mb) antibody β.Antiantibody band 7 bag by sheep anti mouse two anti-.
Referring to Fig. 2, label 21 is wells, the 22nd, and T
1Detect band, the 23rd, T
2Detect band, the 24th, C contrasts band.
The kit that this kind one-step multiple-index mark detects the miocardial infarction mark can have various ways, as: Mb, cTnT joint inspection kit, GPBB, cTnI, CK-MB joint inspection kit etc.Owing to adopted the separation of whole blood film, therefore can be applicable to serum, whole blood simultaneously.In the production technology of kit, the index (as: cTnT etc.) of low concentration has been adopted the technology of biotin amplification, thereby increased the sensitivity that detects.
This kit also is equipped with blood taking needle, little suction pipe and vacuum-packed cotton ball soaked in alcohol in forming, except hospital uses, also applicable to family.
Embodiments of the invention adopt myoglobins (myoglobin, Mb), cardiac troponin (cardiacTroponin T, cTnT) joint-detection gold mark detection kit fast.
Myoglobins (myoglobin, Mb) and cardiac troponin (cardiac Troponin T cTnT) is two kinds of materials that the impaired back of cardiac muscle discharges, wherein Mb is early stage releaser, its critical value is 75ng/ml, and cTnT is the special mark of tool of miocardial infarction, and its critical value is 0.1ng/ml.Detect in the time of for these two kinds of indexs and can improve sensitivity and specificity.This kit is put in these two kinds of detections on the check-out console, and only need 2 are bled and can be carried out two detections simultaneously, and are fast and convenient and accurate.
Referring to Fig. 3, zero expression collaurum among the figure, ↓ expression biotin, label 31 is for having connect the cTnT monoclonal antibody a of collaurum, 32 for having connect the Mb monoclonal antibody α of collaurum, and 33 for having connect the cTnT monoclonal antibody b of biotin, and 34 for being coated on the Streptavidin on the film, 35 for being coated on the Mb monoclonal antibody β on the film, and 36 is that the sheep anti mouse two that is coated on the film resists.
This kit adopts principles such as separation of whole blood system, double-antibody sandwich immune detection system, the amplification of biotin signal, and it is as follows that it detects principle:
1, after dripping whole blood on the sample pad, by the separation of separation of whole blood film, serum is separated to come, and further moves to the collaurum pad.
2, in the Mb detection system, the Mb that exists in the anti-Mb monoclonal antibody α that connects collaurum and the blood specimen combines formation antigen-antibody complex chromatography and moves to detection zone T
1The place combines aubergine band of formation with the anti-Mb monoclonal antibody β that is coated on this.
3, in the cTnT detection system, the anti-cTnT monoclonal antibody a of connection collaurum combines with the cTnT in the blood specimen and forms antigen-antibody complex.This compound again incorporation of markings the anti-cTnT monoclonal antibody b of biotin, form another aubergine band, move to detection zone T through chromatography
2The place is embedded in this Streptavidin and catches fixing.
4, simultaneously, unnecessary Mb monoclonal antibody α-collaurum and cTnT monoclonal antibody a-collaurum move to the check plot, with aubergine C band of sheep anti mouse two anti-reaction generations herein.
Comprise in each packing of this kit: 1 of check-out console, drying agent 1g, 1 of blood taking needle, 1 in little suction pipe, vacuum inner wrapping sterilized cotton ball 1 bag, 1 part of operation instructions, 1 of external packing aluminium foil bag.Under the situation of Packing Sound, under 2 ℃～28 ℃ condition, to preserve, storage life is 12 months.
During use, need check-out console is returned to room temperature; Tear external packing, take out each component, nameless terminal with the sterilized cotton ball wiping, with the blood sampling pinprick once; Get blood with little suction pipe, drip 2 in the sample well of check-out console; Observations after spending 10 minutes.
The detection of conventional miocardial infarction needs the variation according to each index of different time, so the tester is uncomfortable from the chest just should detect, after 2～4 hours again survey once, the variation of Mb and cTnT concentration in the observation blood.
Judged result is as follows:
Positive findings is: T
1The aubergine band appears in the district, and Mb concentration＞75ng/ml is described; T
2The aubergine band appears in the district, and cTnT concentration＞0.1ng/ml is described.
Negative findings is: T
1District, T
2Qu Jun does not have band and occurs.
Null result is: if the no colour band in C district, then testing result is invalid, and sample should detect again.
Referring to Fig. 4, listed the situation of testing result in the table.
Claims (3)
1. the quick gold of an one-step multiple-index mark detection miocardial infarction is marked detection kit, it comprises base plate, the sample pad that is provided with in the base plate lower end, the thieving paper that is provided with in the base plate upper end, and at the glass fibre membrane of locating to post above the sample pad that is fixed with golden labeling antibody that is positioned at of base plate, being sprayed with anti-A antibody II band in the zone between glass fibre membrane and upper end thieving paper of described base plate respectively promptly wraps by myoglobins antibody β and antiantibody band, it is characterized in that, also be sprayed with the Streptavidin band in the zone between glass fibre membrane and upper end thieving paper of described base plate; The golden labeling antibody of described glass fibre membrane bag quilt comprises gold mark anti-A antibody I and gold mark anti-B antibody I, gold mark anti-A antibody I is anti-myoglobins antibody α, gold mark anti-B antibody I is anti-cardiac troponin antibody a, glass fibre membrane also wraps the anti-B antibody II that has been coupled biotin, and anti-B antibody II is anti-cardiac troponin antibody b; This kit adopts the separation of whole blood system, and after dripping whole blood on the sample pad, by the separation of separation of whole blood film, serum is separated to come, and further moves to the collaurum pad.
2. one-step multiple-index mark according to claim 1 detects the quick gold mark detection kit of miocardial infarction, it is characterized in that the composition of this kit also comprises check-out console, drying agent, blood taking needle, little suction pipe and vacuum inner wrapping sterilized cotton ball.
3. one-step multiple-index according to claim 1 and 2 mark detects the quick gold mark detection kit of miocardial infarction, it is characterized in that, described antiantibody band bag by sheep anti mouse two anti-.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011130237A CN1166953C (en) | 2001-05-30 | 2001-05-30 | Fast gold mark test reagent kit for one-step multiple-index test of myocardial infarction |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011130237A CN1166953C (en) | 2001-05-30 | 2001-05-30 | Fast gold mark test reagent kit for one-step multiple-index test of myocardial infarction |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1335502A CN1335502A (en) | 2002-02-13 |
| CN1166953C true CN1166953C (en) | 2004-09-15 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB011130237A Expired - Lifetime CN1166953C (en) | 2001-05-30 | 2001-05-30 | Fast gold mark test reagent kit for one-step multiple-index test of myocardial infarction |
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| Country | Link |
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| CN (1) | CN1166953C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102841207A (en) * | 2011-06-24 | 2012-12-26 | 北京乐普医疗科技有限责任公司 | Fluorescent immune chromatographic test strip for quantitively detecting troponin I and preparation method thereof |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1628136A1 (en) * | 2004-08-19 | 2006-02-22 | B.R.A.H.M.S. Aktiengesellschaft | Method of diagnosis of disease using copeptin |
| CN102103145B (en) * | 2011-02-24 | 2013-01-16 | 南京基蛋生物科技有限公司 | Colloidal gold test strip for double-amplification system and preparation method thereof |
| CN102103143B (en) * | 2011-02-24 | 2013-09-25 | 南京基蛋生物科技有限公司 | Colloidal gold test strip for double-index detection and preparation method thereof |
| CN102323422B (en) * | 2011-05-30 | 2014-03-12 | 中国科学院上海微系统与信息技术研究所 | Immunochromatographic test strip for semi-quantitatively and simultaneously detecting cTnI and Myo and preparation method thereof |
| CN104833798B (en) * | 2015-04-27 | 2016-08-24 | 杭州金溪生物技术有限公司 | Quick diagnosis test strips based on homogeneous chemistry luminescence technology |
| CN110531040A (en) * | 2018-05-25 | 2019-12-03 | 江苏万略医药科技有限公司 | A kind of drug sample analysis system of clinical drug trial |
-
2001
- 2001-05-30 CN CNB011130237A patent/CN1166953C/en not_active Expired - Lifetime
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102841207A (en) * | 2011-06-24 | 2012-12-26 | 北京乐普医疗科技有限责任公司 | Fluorescent immune chromatographic test strip for quantitively detecting troponin I and preparation method thereof |
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| Publication number | Publication date |
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| CN1335502A (en) | 2002-02-13 |
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