CN103852578B - The detachable lath of ELISA Plate - Google Patents
The detachable lath of ELISA Plate Download PDFInfo
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- CN103852578B CN103852578B CN201410015886.7A CN201410015886A CN103852578B CN 103852578 B CN103852578 B CN 103852578B CN 201410015886 A CN201410015886 A CN 201410015886A CN 103852578 B CN103852578 B CN 103852578B
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
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Abstract
The invention provides the detachable lath of ELISA Plate, it significantly can reduce the cost that enzyme linked immunosorbent detection is analyzed, and realization response cup is integrated with reagent cup, is convenient to simplify semi-automatic and full-automatic instrumentation.It is as the solid phase carrier of the analytical control of clinical or basic immunology, biological chemistry and microorganism, comprise some holes cup, it is characterized in that: hole cup comprises reagent cup and reaction cup, identical or the different reaction reagents needed for analysis detection is filled with in advance in each reagent cup, the opening of reagent cup is by thin-film package, and lath can coordinate with 96 hole enzyme target grillages of standard to be installed.
Description
Technical field
The present invention relates to the ELISA Plate that enzyme-linked immuno assay detection method uses, particularly relate to the detachable lath of ELISA Plate.
Background technology
Over nearly 20 years, the enzyme-linked immuno assay technology (ELISA) based on chromogenic substrate, chemical luminous substrate and fluorogenic substrate is widely used in clinical and basic immunology, biological chemistry and microbiological art, has become routine techniques means.The full-automation realized based on the operating process of the elisa assay of 96 hole ELISA Plate at present or semi-automation, and relevant automation equipment obtains in the clinical labororatory of hospital and the research institution such as university, scientific research institutions and promotes widely.Semi-automatic or full-automaticization operation can significantly improve the accuracy of analysis and the repeatability of result, reduces result variation (deviation) that operator brings, improves the flux analyzed.
Carry out the hole cup that elisa assay needs to be coated with in advance bioactive molecule, wrap and generally existed with the form of 96 hole ELISA Plate by hole cup, need reactant liquor, dilution, enzyme labeling liquid, the solution such as signal substrate and cleaning fluid is for completing of reacting simultaneously.Therefore general automatical analysis instrument all will arrange solution region, for placing these solution, and in the cup of hole, drips solution on request by automatic moving liquid device, therefore the general more complicated of full-automatic instrument device structure.Simultaneously, due to daily carry out elisa assay time, especially in clinical labororatory, often need to analyze multiple test item simultaneously, and enzyme mark thing required for each test item, reactant liquor are all not identical, so an automation mechanized operation instrument often has tens reagent positions, this situation further increases again structure to self-reacting device and operation requirements, and the complexity of instrument is improved further.This be suitable for multiple test item half, full-automatic EILISA analytical instrument is expensive.
In addition, because existing elisa assay reagent is all exist with the form of 96 orifice plates, namely 96 samples can once be analyzed in theory, so in the less laboratory of sample size, traditional ELISA reagent is not very convenient to use, and likely causes the waste of reagent, increases analysis cost.In middle-size and small-size clinical labororatory, or the basic science laboratory that sample size is little, existing robotization elisa assay detection technique platform often can not be suitable for, and needs badly and have strategic structural in methodology.How to solve at the semi-automatic or automatical analysis of the less labs of sample size based on multiple analysis projects of 96 orifice plate ELISA, also just become one of striving direction in the field at present.
Summary of the invention
For the problems referred to above, the invention provides the detachable lath of ELISA Plate, it significantly can reduce the cost that enzyme linked immunosorbent detection is analyzed.
Its technical scheme is such, the detachable lath of ELISA Plate, it is as the solid phase carrier of the analytical control of clinical or basic immunology, biological chemistry and microorganism, comprise some holes cup, it is characterized in that: described hole cup comprises reagent cup and reaction cup, realization response cup is integrated with reagent cup, identical or the different reaction reagents needed for analysis detection is filled with in advance in each reagent cup, the opening of described reagent cup is by thin-film package, and described lath can coordinate with 96 hole enzyme target grillages of standard to be installed.
It is further characterized in that:
Described lath is 8 hole cup laths or 12 hole cup laths, and in 8 hole cup laths, the quantity of reaction cup is 1 to 4, and in 12 orifice plate bars, the quantity of reaction cup is 1 to 6;
It also comprises mount pad, and it is one overall and be fixed in described mount pad side that some reagent cup connect successively, and described mount pad is provided with cell body, and described reaction cup is installed in described cell body;
The front and rear sides inwall of described cell body is provided with draw-in groove symmetrically, and the group number of described draw-in groove is equal with the number of described reaction cup, and the front and rear sides of described reaction cup is engaged by described draw-in groove.
It is further characterised in that:
Described draw-in groove is provided with some groups, is provided with the dividing plate of the front and rear sides inwall supporting described cell body between adjacent two groups of draw-in grooves;
Symmetrically on the arranged on left and right sides inwall of described cell body and on the arranged on left and right sides end face of described dividing plate be provided with fixture block, the arranged on left and right sides of described reaction cup engages with described fixture block;
Described reaction cup is cylindricality or back taper, and described fixture block is positioned at described dividing plate and described cell body bottom, and the inner face of described draw-in groove and described fixture block is circular arc camber;
Coated layer is provided with in described reaction cup;
The top of described reaction cup is provided with radial flange;
The upper/lower terminal of described cell body is provided with opening, and the lower ending opening of described cell body is provided with blend stop, and described blend stop is provided with cushion block;
Described reagent wells cup can the filling reaction reagent corresponding for different test items in advance, as enzyme marker, biotin labeled antibody or sample diluting liquid.
Beneficial effect of the present invention is:
(1) by the integrated design of reaction cup and reagent cup, and reagent needed for enzyme linked immunosorbent detection is filled with in advance in reagent wells, not only reduce or avoid the design of reagent position of automation mechanized operation instrument, significantly simplify its structure, and reduce its operation requirements, be convenient to simplify semi-automatic and full-automatic instrumentation, and reduce the manufacturing cost of automation mechanized operation instrument;
(2) detected while multiple test items of a small amount of sample by 8 single holes or the realization of 12 orifice plate bars, not only avoid the problem that traditional 96 hole ELISA Plate utilization factors are low, reduce the use cost of ELISA Plate, and significantly simplify the running program of operator and equipment, improve the detection speed of sample, reduce the human cost of detection, and realize the needs that beside sickbed detects (POCT) fast.
(3) reaction cup is dismountablely be installed on the mount pad of lath, can avoid scrapping of the whole lath caused because of single reaction cup application of sample mistake, can reduce the use cost of ELISA Plate further; Arranging of draw-in groove and fixture block can effective fixation reaction cup, dividing plate the intensity and stability that can increase mount pad are set, the top of reaction cup is provided with radial flange, conveniently holds.
Accompanying drawing explanation
Fig. 1 is structural representation of the present invention;
Fig. 2 is the plan structure schematic diagram of Fig. 1;
Fig. 3 is the structural representation of mount pad of the present invention;
Fig. 4 is the assembling schematic diagram of the sheet frame of the present invention and existing 96 hole ELISA Plate.
Embodiment
As shown in Figure 1 to Figure 3, the detachable lath of ELISA Plate, it is as clinical or basic immunology, the solid phase carrier of the analytical control of biological chemistry and microorganism, lath can be assemblied in 96 hole enzyme target grillages of standard, comprise 8 hole cups, hole cup comprises reagent cup 1 and reaction cup 2, the quantity of reaction cup 2 is 3, Fig. 1, in Fig. 2, one of them reaction cup 2 is not shown, be filled with in advance respectively in each reagent cup 1 and analyze the reaction reagent 13 needed for detecting, 14, 15, 16, 17, reagent 13, 14, 15, 16, 17 are selected from enzyme marker, biotin labeled antibody or sample diluting liquid, enzyme marker can be enzymic-labelled antibody or enzyme-labelled antigen, the opening of reagent cup 1 is encapsulated by film 12.
It also comprises mount pad 3, and it is one overall and be fixed in mount pad 3 side that 5 reagent cup 1 connect successively, and mount pad 3 is provided with cell body 18, and reaction cup 2 is installed in cell body 18; The front and rear sides inwall of cell body 18 is provided with draw-in groove 4 symmetrically, and the group number of draw-in groove 4 is equal with the number of reaction cup 2, and the front and rear sides of reaction cup 2 is engaged by draw-in groove 4; Draw-in groove 4 is provided with some groups, is provided with the dividing plate 8 of the front and rear sides inwall supporting cell body 18 between adjacent two groups of draw-in grooves 4; Symmetrically on the inwall of cell body 18 arranged on left and right sides and on the arranged on left and right sides end face of dividing plate 8 be provided with fixture block 5, the arranged on left and right sides of reaction cup 2 engages with fixture block 5; Reaction cup 2 is cylindricality, and fixture block 5 is positioned at dividing plate 8 and cell body 18 bottom, and the inner face of draw-in groove 4 and fixture block 5 is circular arc camber; Coated layer is provided with in reaction cup; The top of reaction cup 2 is provided with radial flange 9; The upper/lower terminal of cell body 18 is provided with opening, and the lower ending opening of cell body 18 is provided with blend stop 6, and blend stop 6 to be provided with in cushion block 7, figure the holding part that 10,11 are lath two ends.
As shown in Figure 4, the present invention can be assemblied on the sheet frame of existing 96 hole ELISA Plate, the number of lath can be set according to detection demand, significantly improve the utilization factor of ELISA Plate, and reacting hole cup quantity and reagent wells cup quantity can be distributed according to the needs of analysis design mothod, also can leave room on lath, the quantity of hole cup is reduced, and the structure of hole cup and size can experimentally need to carry out necessary adjustment.
Elisa assay based on lath of the present invention detects, and may be used for manual operations, but is mainly used in relevant semi-automatic, automatical analysis process.In the operating process of robotization, relative to traditional analytical equipment based on 96 orifice plates, due to not at needs reagent bottle, the requirement to self-reacting device significantly can be simplified.
Following is clinical immunoassays application example based on the principle of the invention.
Example one: pregnant guarantor 4 detection
The routine clinical health check-up big event of pregnant woman comprises folic acid (FA), alpha-fetoprotein (AFP), human chorionic gonadotrophin (HCG), free estradiol (uE2) etc.Folic acid is formed with vital role for nerviduct, and period of gestation lacks the growth that folic acid can affect fetal central nervous system, and the folic acid that therefore pregnant woman's absorption before conceived is enough is very important for the normal development of fetus.Research showed to supplement the birth rate that enough folic acid can reduce neural tube defects and harelip fetus in pregnancy early stage and pregnancy.Down syndrome to make a variation the phenomenon caused to chromosomal three bodies by the 21st of human body the, and this is also the disease that the chromosome deficiency of mankind's Late Cambrian causes.China's year birth neonate is about 1,600 ten thousand, and the birth rate of Down Syndrome is 1/800 ~ 1/600, then annual appointment increases Tang Shi more than 20000 name.Current clinical conventional immunological method detects the technological means of Down's syndrome for getting person under inspection's venous blood and separation of serum, measure alpha-fetoprotein (AFP), FE3 (uE3) and human chorionic gonadotrophin (HCG) content in serum, and according to person under inspection's age, body weight, pregnant week and be whether that the factors such as diabetic carry out calculatings MMOL value by mathematical analysis examination software and relative risk is assessed.Current method of testing is clinically apply 4 kits respectively to carry out independent detection analysis to a serum sample, and operation steps is many, is unfavorable for practical application, and is not suitable for the detection of few this project of labs of sample size.
Adopt principle of the present invention, all reagent of above-mentioned 4 test items and bag can be placed on one by hole and analyze on lath, form shown in its structural table 1 of comprehensive detection for a sample.
Table 1: folic acid (FA), alpha-fetoprotein (AFP), human chorionic gonadotrophin (HCG), the plank designs example that free estradiol (uE2) pregnant guarantor's tetrad enzyme joint inspection is surveyed
Folic acid testing process needs sample preprocessing, so reaction reagent comprises 3, Cleaning Principle is Immune competition method, and wrapping by hole is FABP, and reagent 1 and 2 is sample treatment liquid, and reagent 3 is folic acid enzyme marking reagent;
AFP and HCG detection mode is immunoassay sandwich method, and reagent 1 is biotinylated first antibody reagent, and reagent 2 is ELIAS secondary antibody reagent, AFP and HCG bag is that Avidin bag is by hole by hole;
UE2 is that Small molecular detects, and adopt Immune competition method, uE2 reaction reagent is E2 enzyme marking reagent, is at war with the free E2 in sample, and wrapping by hole is anti-E2 antibody;
Above-mentioned four link detection reagents can be carried out on semi-automatic or automatical analysis equipment, and testing conditions can be optimized respectively according to the feature of biomaterial, and maximum response time is no more than 30 minutes, and temperature of reaction is 25-37
0c, sample size is 10 microlitre-100 microlitres.
Example two: the two-in-one detection of end Natriuretic Peptide/cardiac muscle troponin I
NT-proBNP is the N end fragment of non-activity after the division of BNP prohormone, is mainly secreted by left ventricle when cardiac muscle cell is subject to volume load and pressure load increases.It is incomplete that NT-proBNP can be used for EARLY STAGE EVALUATION cardiac systolic function, and EARLY STAGE EVALUATION Diastolic Heart failure and ventricle wall segmentation movement harmony, have higher susceptibility and negative predictive values.
Troponin by I, T, C tri-subunit form, they together with tropomyosin by regulate Ca
2+modulate actin and myosin interaction are come to the activity of striated muscle filamentous actin ATP enzyme.When after myocardial damage, cardiac troponin complex is discharged in blood, and after 4 ~ 6 hours, start to raise in blood, the cTnI of rising can keep for a long time (6 ~ 10 days) in blood, which provides longer detection period.CTnI has Cardiac-specific, cTnI is all higher to the Sensitivity and Specificity detecting myocardial damage, chronic heart failure can occur that low-level raises, and severe has symptom heart failure to there is myocardium cell necrosis, the continuous disintegration of muscle fibril, the sustainable rising of serum Myocardial Troponin level.
Clinically by the quantitative measurment carrying out above-mentioned two test items simultaneously, realize the quick and precisely diagnosis of miocardial infarction.
Adopt principle of the present invention, all reagent of above-mentioned 2 test items and bag can be placed on one by hole analyzes on lath, form for a sample, comprise the quantitative comprehensive detection of NT-proBNP and cardiac muscle troponin I in serum, blood plasma, whole blood, shown in its structural table 2.
Table 2:N-holds the quantitative comprehensive detection cardiac muscle stalk table 2 of Natriuretic Peptide (NT-proBNP) and cardiac muscle troponin I (cTnI): the plank designs example that the joint inspection of plug bigeminy enzyme is surveyed
The Cleaning Principle of NT-proBNP and cTnI two test items is immuno-sandwich method.Bag is by hole for being coated with Avidin, and reagent 1 is biotinylated first antibody reagent, and reagent 2 is ELIAS secondary antibody reagent, such as horseradish peroxidase (HRP);
Above-mentioned two link detection reagents can be carried out on semi-automatic or automatical analysis equipment, and testing conditions can be optimized respectively according to the feature of biomaterial, and maximum response time is no more than 20 minutes, and temperature of reaction is 35-37
0c.Sample size is 10 microlitre-100 microlitres.
Example three: the joint-detection of omnidistance c reactive protein (HS-CRP and CRP)
C reactive protein (C-Reactiveprotein, be called for short CRP), found early than nineteen thirty, was a kind ofly to form the Acute reaction protein of compound with CPS precursor reactant.The detection of CRP was widely used in clinical as the non-specific markers of inflammation and tissue damage before the eighties, but due to the detection method comparatively backwardness of past CRP, false positive and false negative are very high, have impact on its value clinically, and are ignored by clinical gradually.In recent years, due to the renewal of detection technique, measure quick, the easy and reliable method of CRP and set up rapidly, CRP is increased greatly in clinical practice field, its value medically is just just extensively being tested and is being admitted.
Hs-CRP (HighsensitivityC-reactiveprotein) and CRP are not two kinds of albumen, and just distinguished from sensitivity, hs-CRP (HS-CRP) lowest detectable limit reaches 0.1mg/l; Originally thought that CRP was that normal serum but finds that angiocardiopathy occurs is closely related with following, a large amount of research data shows, it be the process of fat accumulation is also outward a chronic inflammation processes that atheromatous thrombus eliminates; HS-CRP slightly raises relevant to coronary artery events, apoplexy and peripheral angiopathy, is independently hazards; HS-CRP has been proved to be and has caused the independent hazard factor of angiocardiopathy by chronic inflammation, detects its concentration and to play an important role to the intervention of angiocardiopathy and prognosis and by clinical attention.Epidemiology survey also shows, and the probability that acute apoplexy occurs HS-CRP level rising person is 2 times of normal healthy people, and the probability that myocardial infarction occurs is 3 times of normal person.European hypertension prevention and control guide (ESH/ESC) formal recommendation in 2003, hyperpietic need detect HS-CRP level.
What the sensitivity detected due to current CRP did not reach HS-CRP detects requirement, so bring inconvenience to clinical judgment, simultaneously because the detection reagent sensitivity of HS-CRP is higher, often in high CRP situation, reach the plateau detecting reagent, HS-CRP detects reagent can not report correct CRP level.Employing two kinds of reagent of generally having to clinically originally detect respectively to same increment.
The major significance of the clinical detection of CRP and HS-CRP is as follows:
1, CRP raised rapidly as acute phase protein a few hours after the seizures of disease such as various acute inflammation, tissue damage, miocardial infarction, operation wound, radioactive damage, and have the gesture be doubled and redoubled; when pathology takes a turn for the better; be down to rapidly again normal, the degree of its elevation amplitude and infection is proportionate.
2, the correlativity of CRP and other inflammatory factor: CRP and other inflammatory factor such as total white blood cells, erythrocyte sedimentation rate (ESR) and polymorphonuclear leukocyte etc. have Close relation, there is positive correlation in CRP and WBC, in inflammatory reaction, play positive role, make human body have nonspecific resistance.When patient disease is shown effect, CRP can rise early than WBC, replys normal also very fast, therefore has high susceptibility.
3, CRP can be used for the antidiastole of bacterium and virus infections: once be inflamed, and namely CRP level raises, and viral infection CRP is mostly normal; Septicopyemia CRP raises rapidly, and relying on blood culture then at least needs 48 hours, and its positive rate is not high.
Measure clinically by the accurate quantitative analysis carrying out above-mentioned CRP and HS-CRP, thus improve the clinical value of CRP index.
Adopt principle of the present invention, all reagent of above-mentioned CRP test item and bag can be placed on one by hole and analyze on lath, form for a sample, comprise the accurate quantitative analysis comprehensive detection of CRP in serum, blood plasma, whole blood, shown in its structural table 3.
The quantitative comprehensive detection reaction lath design example of table 3:CRP
The Cleaning Principle of CRP test item is immuno-sandwich method, wrap by hole as being coated with Avidin, reagent 1 is biotinylated first antibody reagent, reagent 2 is ELIAS secondary antibody reagent, the second antibody that such as horseradish peroxidase (HRP) marks, sample diluting liquid carries out necessary dilution to sample, and for detecting in rising tone the CRP albumen having higher level, such as CRP concentration is positioned at 0.01mg/L – 10mg/L;
Above-mentioned CRP detects reagent can be carried out on semi-automatic or automatical analysis equipment, and testing conditions can be optimized according to the feature of biomaterial.Maximum response time is no more than 20 minutes.Temperature of reaction is 35-37
0c, sample size is 10 microlitre-100 microlitres.
Claims (9)
1. the detachable lath of ELISA Plate, it is as the solid phase carrier of the analytical control of clinical or basic immunology, biological chemistry and microorganism, comprise some holes cup, it is characterized in that: described hole cup comprises reagent cup and reaction cup, realization response cup is integrated with reagent cup, be filled with the identical or different reaction reagents needed for analysis detection in each reagent cup in advance, the opening of described reagent cup is by thin-film package, and described lath coordinates with 96 hole enzyme target grillages of standard to be installed; Described lath also comprises mount pad, and it is one overall and be fixed in described mount pad side that some reagent cup connect successively, and described mount pad is provided with cell body, and described reaction cup is installed in described cell body.
2. the detachable lath of ELISA Plate according to claim 1, is characterized in that: described lath is 8 hole cup laths or 12 hole cup laths, and in 8 hole cup laths, the quantity of reaction cup is 1 to 4, and in 12 orifice plate bars, the quantity of reaction cup is 1 to 6.
3. the detachable lath of ELISA Plate according to claim 1, it is characterized in that: the front and rear sides inwall of described cell body is provided with draw-in groove symmetrically, the group number of described draw-in groove is equal with the number of described reaction cup, and the front and rear sides of described reaction cup is engaged by described draw-in groove.
4. the detachable lath of ELISA Plate according to claim 3, is characterized in that: described draw-in groove is provided with some groups, is provided with the dividing plate of the front and rear sides inwall supporting described cell body between adjacent two groups of draw-in grooves.
5. the detachable lath of ELISA Plate according to claim 4, is characterized in that: symmetrically on the arranged on left and right sides inwall of described cell body and on the both sides end face of described dividing plate be provided with fixture block, and the arranged on left and right sides of described reaction cup engages with described fixture block.
6. the detachable lath of ELISA Plate according to claim 5, is characterized in that: described reaction cup is cylindricality or back taper, and described fixture block is positioned at described dividing plate and described cell body bottom, and the inner face of described draw-in groove and described fixture block is circular arc camber.
7. the detachable lath of ELISA Plate according to claim 1, is characterized in that: be provided with coated layer in described reaction cup, and the top of described reaction cup is provided with radial flange.
8. the detachable lath of ELISA Plate according to claim 1, it is characterized in that: the upper/lower terminal of described cell body is provided with opening, the lower ending opening of described cell body is provided with blend stop, and described blend stop is provided with cushion block.
9. the detachable lath of ELISA Plate according to claim 1, it is characterized in that: described reagent cup can the filling reaction reagent corresponding for different test items in advance, and described reaction reagent is selected from any one in enzyme marker, biotin labeled antibody or sample diluting liquid.
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| CN109669041A (en) * | 2019-02-02 | 2019-04-23 | 上海微银生物技术有限公司 | A kind of enzyme immune detection lath, enzyme plate for detecting immunity and its detection method |
| CN110530867A (en) * | 2019-10-11 | 2019-12-03 | 江苏贝索生物工程有限公司 | The micro- test tube reaction cup of cassette and the full-automatic test tube detection method for utilizing the reaction cup |
| WO2023122970A1 (en) * | 2021-12-28 | 2023-07-06 | 深圳迈瑞动物医疗科技股份有限公司 | Medical point-of-care testing device |
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| CN202486136U (en) * | 2012-04-09 | 2012-10-10 | 无锡国盛精密模具有限公司 | Enzyme-linked immuno-sorbent assay (ELISA) plate shelf |
| CN202693591U (en) * | 2012-08-13 | 2013-01-23 | 沃克(天津)生物科技有限公司 | Elisa plate easy to dismount |
| CN102866250A (en) * | 2012-09-26 | 2013-01-09 | 无锡耐思生物科技有限公司 | Elisa plate structure |
| CN203732540U (en) * | 2014-01-14 | 2014-07-23 | 白仲虎 | Detachable batten of elisa plate |
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| Publication number | Publication date |
|---|---|
| CN103852578A (en) | 2014-06-11 |
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Application publication date: 20140611 Assignee: Yancheng Baiming Biological Technology Co. Ltd. Assignor: Bai Zhonghu Contract record no.: 2016320010002 Denomination of invention: Detachable batten for enzyme-linked immuno sorbent assay (ELISA) plate Granted publication date: 20151118 License type: Common License Record date: 20160216 |
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