CN2779402Y - Chromatic multi-form test strip for HCG test - Google Patents
Chromatic multi-form test strip for HCG test Download PDFInfo
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- CN2779402Y CN2779402Y CN 200520056329 CN200520056329U CN2779402Y CN 2779402 Y CN2779402 Y CN 2779402Y CN 200520056329 CN200520056329 CN 200520056329 CN 200520056329 U CN200520056329 U CN 200520056329U CN 2779402 Y CN2779402 Y CN 2779402Y
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Abstract
The utility model belongs to the field of medical detection, particularly to a chromatic multi-form HCG test paper strip. The test paper strip is formed by orderly pasting a sample cushion, a glass fiber membrane coated with a gold mark antibody, a coating membrane and absorbent paper on a substrate in a mutually spliced way. The coating membrane is provided with a detection line which coats the HCG antibody and a control line which coats the anti-rat antibody. The coloration colors of the detection line and the control line have diversity, and colloidal gold particles with different colors are obtained by controlling the size of the colloidal gold particle or adopting emulsion particles with different colors. The coloration forms of the detection line and the control line have diversity, and the detection line and the control line are sprayed into various strip belts with different shapes, such as a strip shape, a rhombus shape, a circle shape, a quincunx shape and a cross shape, by adjusting the set program of the membrane jetter in the manufacturing technique. The utility model has abundant colors of banding and various forms, so the utility model can detect the HCG content in urine more exactly and meet the demand of different people. The utility model is applied to the self test of families and individuals more extensively.
Description
Technical field
The utility model belongs to biomedicine field, and particularly the medical test technology specifically relates to the multi-form HCG test paper of a kind of colour bar.
Background technology
The mature women forms the embryo because of the ovum of fertilization moves in the uterine cavity behind the implantation, is in the fetus process at growing, and placenta plamoditrophoblast cell produces a large amount of human chorionic gonadotrophins (HCG), can be excreted in the urine by the pregnant woman blood circulation.HCG began secretion in 10～14 days in the back of becoming pregnant, and can rise to greater than 2500IU/L in 35-50 days, reached secretion peak 8 hundred million IU/L in 60～70 days, was the sensitive index of early pregnancy.Meanwhile, along with the development of modern medicine, people find that more and more the content of HCG and many neoplastic disease have close ties, when taking place as the malignant tumour of chorioepithelioma and reproductive system, unusual HCG can occur in the urine.The improper gestation of other, as ectopic pregnancy, HCG was unusual during vesicular mole etc. also can cause urinating.Therefore the accurate detection of HCG concentration in urine not only can be used as early pregnancy diagnosis, also can be used as a kind of aided detection method of neoplastic disease.Lag behind under the external situation at present at complicated self-reacting device of China and matched reagent thereof, development and production special easy, the HCG testing product all has a wide range of applications and wide development space to diagnosis, discriminating, treatment and the tracing study etc. of early pregnancy, pathology gestation duty ectopic pregnancy and placental trophoblast disease fast.
The diagnosis that HCG is unusual is a standard with the HCG that detects in the urine mainly, and its inspection method is based on immunization.Present detection method both domestic and external mainly comprises: latex or hemagglutination-inhibition test, radio-immunity, enzyme immunity, colloidal gold method etc.This several method following characteristics of having compared.
Latex agglutination inhibiting test (latex agglutination inhibitiontest, LAIT) and the hemagglutination-inhibition test of haemocyte (hemoagglutination inhibition test, HAIT) these two kinds tests are convenient easy, sensitivity is 100-500mU/ml, be fit to large quantities of sample inspections, but because of poor specificity, can not be quantitative, be substituted gradually.
Radioimmunoassay (RIA) needs a locking equipment, and procedure of test is loaded down with trivial details, and the nucleic pollution problem is arranged, and is not suitable for routine clinical application.
Enzyme linked immunosorbent assay (ELISA) running program is complicated, easily causes false positive and false negative, and easily causes the operator to be injured and environmental pollution, and test period is longer, needs just can draw testing result more than two hours.Must possess simultaneously microplate reader and wash the plate machine, this basic unit laboratory and small-sized outpatient service difficulty reach, and test is subjected to condition restriction such as temperature, makes troubles to detection.
And immune-gold labeled law technology (Immunochromatography Assay) grows up the early 1990s, but because of it is quick, easy and simple to handle, the accumulating of stable reagent room temperature, be difficult for pollution characteristics and be widely used.It is the combination of immune affine technology, engram technology, immunolabelling technique and chromatographic technique.With coated antibody, colloid gold label antibody immobilization is combined with sample sorbing material etc., is prepared as immunochromatography diagnostic test/plate, only need be during use inserting sample solution under the test strips, and several minutes just can judged result.With the immunity percolation method relatively, good stability operate easylier, quick, and owing to be dried strip/test plate (panel) form, need not cryopreservation, accumulating makes things convenient for.
At present, the existing product that utilizes immune colloidal gold technique to detect HCG has following shortcoming:
1. adopt the method for 4 bands colour developing, relatively poor for the sample specificity of high concentration.
2. it is single to show band color and form.
Summary of the invention
The purpose of this utility model provides the multi-form HCG test paper of a kind of colour bar, it adopts the colloidal gold chromatography preparation of two bands colour developing, show band color abundant (red, orchid, black, purple etc.), various informative (stripe shape, plus sige shape, circular point shape, quincunx etc.), thereby its demand that can more accurately detect the content of HCG in the urine and can satisfy different crowd, be widely used in family and individual self check more.
The multi-form HCG test paper of colour described in the utility model bar, be that mutual in turn overlap joint ground is pasted glass fibre membrane, coated film and the thieving paper of sample pad, coated with gold labeling antibody and formed on end liner, coated film has detection line and control line, the detection line bag is by HCG antibody, and the control line bag is by anti-mouse antibody; The colour developing color of described detection line and control line has diversity, is the size by the control colloid gold particle, obtains the colloid gold particle of different colours, perhaps adopts the emulsion particle of different colours; The colour developing form of described detection line and control line has diversity, is that spray is bar shaped, rhombus, circle, quincunx, the multiple difform band of cruciform by the setting program of Membrane jetter in the adjusting production technology.
Described present latex particulate can be from abroad buying as companies such as magsphere.
The HCG antibody of described bag quilt is highly purified anti-monoclonal antibody and/or polyclonal antibody, the anti-β of mark-HCG monoclonal antibody on described colored colloid gold particle or the colored latex particle.
The preparation method of the multi-form HCG test paper of colour described in the utility model bar may further comprise the steps:
A. the preparation of coated film:
Detection line: the debugging Membrane jetter, spouting liquid is 20 microlitres/35 centimetre, is cushioned liquid dilution HCG monoclonal antibody with bag, and concentration is 80 μ g/ml, and machine is rule, room temperature airing 20 minutes;
Control line: the debugging Membrane jetter, spouting liquid is 20 microlitres/35 centimetre, is cushioned liquid with bag and dilutes anti-mouse IgG polyclonal antibody, concentration is 2mg/ml, the machine line, line-to-line should be careful even every 5mm, room temperature airing 20 minutes.37 ℃ were sealed 60 minutes, took out rearmounted 37 ℃ of oven dry and handled two hours, and envelope is standby;
B. the preparation of golden labeling antibody: transfer collaurum pH value to 7.5 with 0.1M sal tartari, add anti-β-HCG monoclonal antibody by 10 micrograms antibody/milliliter collaurum, mixing left standstill 30 minutes, centrifugal 30 minutes of 13000rpm, abandon supernatant, precipitation mark cleansing solution washed twice, last supernatant discarded will precipitate with the golden labeling antibody of 1/10th initial collaurum volumes and preserve the liquid dissolving, put 4 ℃ standby, one week of effect phase;
The golden labeling antibody that C. will prepare is coated on glass fibre membrane or the polyester film;
The glass fibre membrane of described coated with gold labeling antibody can be marked antigen with gold with commonsense method and be coated on the glass fibre membrane, also can obtain by following method: the collaurum that mark is good is layered on the glass fibre membrane equably, 10 square centimeters of every ml soln shops, freeze drying, envelope, put 4 ℃ standby.
D. on end liner, paste glass fibre membrane, coated film and the thieving paper of sample pad, coated with gold mark antigen in turn mutually overlap joint, obtain test paper plate, cut into the test strips of different in width as requested.
Prescription and preparation method that described bag is cushioned liquid and confining liquid are the general technology in this area.
The utility model adopts double antibody sandwich method, can be used to detect the female urine fluid samples.Compare with existing detection kit,, make it can produce multiple color because golden labeling antibody is carried out particular processing; Simultaneously detection line (T line) and control line (C line) have been carried out particular processing, made it can produce various test format, as bar shaped, circle, plus sige shape, quincunx etc.Described detection line and control line are that spray becomes difform band, thereby satisfies different people, all ages and classes layer to the multifarious demand of test format by the setting program of Membrane jetter in the adjusting production technology.
The utility model utilizes modern international state-of-the-art collaurum and latex immunochromatography technique, provides a kind of hospital or individual family of being used for to test oneself and wait the fast diagnosis reagent of detection very early pregnancy.This reagent has advantages such as high degree of specificity, sensitivity, detection speed be fast, and it is abundant particularly to show the band color, various informative characteristics.And need not use instrument and equipment, with low cost, easy and simple to handle, can be widely used in hospital, family and individual detection for HCG.This test strips can directly be used, and also can be assemblied in the plastic device to use with check-out console, detecting pen form.
Description of drawings
Fig. 1 is the structural representation of test strips described in the utility model.
Fig. 2 is the testing result synoptic diagram.
Fig. 3 is the testing result synoptic diagram.
Embodiment
The multi-form HCG test paper of colour described in the utility model bar, as shown in Figure 1, this test strips is that mutual in turn overlap joint ground is pasted glass fibre membrane 3, coated film 4 and the thieving paper 5 of sample pad 2, coated with gold labeling antibody and formed on end liner 1, coated film 4 has detection line 6 and control line 7, detection line 6 bags are by HCG antibody, and control line 7 bags are by anti-mouse antibody; The colour developing color of described detection line 6 and control line 7 has diversity, is the size by the control colloid gold particle, obtains the colloid gold particle of different colours, perhaps adopts the emulsion particle of different colours; The colour developing form of described detection line 6 and control line 7 has diversity, is that spray is bar shaped, rhombus, circle, quincunx, the multiple difform band of cruciform by the setting program of Membrane jetter in the adjusting production technology.
Manufacture method is as follows:
A antigen preparation: adopt highly purified anti-β-HCG monoclonal antibody as envelope antigen.
The preparation of B antigenic membrane:
Bag is cushioned the preparation of liquid: 0.05M pH9.6 carbonate buffer solution for bag by solution, 0.22 μ membrane filtration mistake, put 4 ℃ standby, one week of effect phase.
The preparation of confining liquid: the preparation 0.01M pH7.0 phosphate buffer (PBS), 0.22 μ membrane filtration mistake, put 4 ℃ standby, one week of effect phase.Preparation sealing working fluid: 2%BSA, 2% skimmed milk, 0.01M pH7.0PBS, 0.22 μ membrane filtration mistake, put 4 ℃ standby, three days effect phases.
Detection line preparation: debugging Membrane jetter, spouting liquid are 20 microlitres/35 centimetre, are cushioned liquid dilution HCG monoclonal antibody with bag, and concentration is 80 μ g/ml, and machine is rule, room temperature airing 20 minutes.
Control line preparation: debugging Membrane jetter, spouting liquid are 20 microlitres/35 centimetre, are cushioned liquid with bag and dilute anti-mouse IgG polyclonal antibody, and concentration is 2mg/ml, the machine line, and line-to-line should be careful even every 5mm, room temperature airing 20 minutes.37 ℃ were sealed 60 minutes, took out rearmounted 37 ℃ of oven dry and handled two hours, and envelope is standby.
The manufacturing rules of C labeling antibody
The preparation of gold chloride: with distilled water dissolved chlorine auric acid, be made into 1% solution, put 4 ℃ standby, three days effect phases.1000ml 1% chlorauric acid solution prescription: 10g gold chloride; The 1000ml distilled water.
The preparation of trisodium citrate: dissolve trisodium citrate with distilled water, be made into 1% solution, put 4 ℃ standby, three days effect phases.
1000ml 1% citric acid three sodium solution prescription: 10g trisodium citrate; The 1000ml distilled water.
0.1M the preparation of sal tartari: with distilled water preparation, 0.22 μ membrane filtration mistake, put 4 ℃ standby, one week of effect phase.1000ml 0.1M solution of potassium carbonate prescription: 13.8g sal tartari; The 1000ml distilled water.
The preparation of 3%PEG-20000: with distilled water preparation, 0.22 μ membrane filtration mistake, put 4 ℃ standby, one week of effect phase.1000ml 3%PEG solution formula: 30g PEG-20000; The 1000ml distilled water.
The preparation of mark cleansing solution: 2%BSA, 0.01M pH7.0PBS solution, 0.22 μ membrane filtration mistake, put 4 ℃ standby, two weeks of effect phase.1000ml mark cleansing solution prescription: 20gBSA; 1000ml0.01M pH7.0PBS solution.
The preparation that the gold labeling antibody is preserved liquid: 1%BSA, 0.5% skimmed milk, 5/0,000 NaN3,0.1%Tween-20,0.01MpH7.0 PBS solution, 0.22 μ membrane filtration mistake, put 4 ℃ standby, 15 days effect phases.1000ml gold labeling antibody is preserved formula of liquid: 10g BSA; The 5g skimmed milk; 0.5gNaN3; 1ml Tween-20; 1000ml0.01M pH7.0PBS solution.
Firing of collaurum: 1% gold chloride is diluted to 0.01% with distilled water, put electric furnace and boil, add 4 milliliter of 1% trisodium citrate, continue to boil by per 100 milliliter of 0.01% gold chloride, be shiny red up to liquid and promptly stop heating, supply dehydration after being cooled to room temperature.Outward appearance should be pure, and is bright, do not have precipitation and floating thing.
The preparation of gold labeling antibody: transfer collaurum pH value to 7.5 with 0.1M sal tartari, add anti-β-HCG monoclonal antibody by 10 micrograms antibody/milliliter collaurum, mixing left standstill 30 minutes, centrifugal 30 minutes of 13000rpm, abandon supernatant, precipitation mark cleansing solution washed twice, last supernatant discarded will precipitate with the golden labeling antibody of 1/10th initial collaurum volumes and preserve the liquid dissolving, put 4 ℃ standby, one week of effect phase.
The drying of D gold labeling antibody
The collaurum that mark is good is layered on the glass fibre membrane equably, 10 square centimeters of every ml soln shops, drying, envelope, put 4 ℃ standby.
Determining of the glass fibre membrane width of E Jin Biaokangtibao quilt
The collaurum glass fibre membrane is cut into 4 millimeters, 5 millimeters, 6 millimeters specifications, forms test-strips with coated film, every wide 4 millimeters.Selection can detect indoor Quality Control sample, and the gold mark glass fibre membrane width of no nonspecific reaction is decided to be the use width.
Cutting of F gold labeling antibody glass fibre membrane
Join the width that experiment is determined according to dripping, golden labeling antibody glass fibre membrane is cut, put between drying shed standby.
Cutting of G thieving paper
With trimmer thieving paper is cut into 35 centimeter length, 4.2 centimetres wide, puts between drying shed standby.
Cutting of H glass fibre membrane
Glass fibre membrane is cut into long 35 centimetres, and wide 2 centimetres is rectangular, puts between drying shed standby.
The stickup of the big plate of I
On request coated film, thieving paper, freeze-drying gold labeling antibody, glass fibre membrane are sticked on the plastic bottom board, form big plate.Composing room's temperature is controlled at 25 ℃, humidity 20%-30%.
The J slitting
With cutting cutter big plate is cut into single person-portion, 3 millimeters of everyone part width, sampling observation at random, sensitivity can detect indoor Quality Control, and band colour developing degree reaches one "+" number, and specific band can pass through the Panel of the Ministry of Public Health nothing but.
K encapsulation and group box
The test strips and one that 100 person-portions have been cut the drying prescription of being responsible for a task until it is completed is encapsulated in the aluminium foil polybag, puts into kit, a instructions of every box.Keep in Dark Place in 4-30 ℃, must not be frozen.
The multi-form HCG test paper of colour described in the utility model bar adopts highly purified anti-β-HCG monoclonal antibody to carry out special HCG as envelope antigen and checks.Its principle is to contain equally distributed colloid gold label β-HCG monoclonal antibody in the test strips, anti-β-HCG antibody and anti-mouse IgG antibody are individually fixed on the nitryl film and are two lines and is arranged above and below, the negative up contrast of anti-mouse IgG antibody, anti-α-HCG antibody below be detection line.When containing HCG antigen in the tested urine sample, the β-HCG antibody of β-HCG elder generation and golden mark carries out specific adsorption, forms the Ag-Ab beta composite.Because the capillarity and the chromatography effect of miillpore filter, the reaction compound moves forward along the nitryl film, when waiting to walk to anti-α-HCG antibody line, forms Ab α-Ag-Ab β-Au compound and is enriched in bag by on the line, red precipitate line of final formation is to be the HCG positive reaction.And irrelevant golden mark mouse IgG continues to go upward to anti-mouse IgG antibody place with urine on the test strips, and the antigen antibody complex that forms red-label with it is negative contrast.
When being used to detect, observing the colour developing and the shape of detection line 6 and control line 7 and judge positive still negative.Shown in Fig. 2 (a1), when detection line 6 and control line 7 all developed the color, the result was positive; Shown in Fig. 2 (a2), when control line 7 colour developing and detection line 6 when not developing the color, the result is negative.
The colour developing form of described detection line 6 and control line 7 has diversity, is that spray is bar shaped, circle, quincunx, the multiple difform band of cruciform by the setting program of Membrane jetter in the adjusting production technology.Shown in Fig. 2 (a1) and Fig. 2 (a2), the colour developing of detection line 6 and control line 7 is bar shaped; Shown in Fig. 2 (b1) and Fig. 2 (b2), the colour developing of detection line 6 and control line 7 is rhombus, circle, quincunx.
Shown in Fig. 3 (a), the cruciform band appears, and promptly detection line 6 and control line 7 intersections are cruciform, and the result is positive; Shown in Fig. 3 (b), only show belt strips at control line 7, the result is negative.
Claims (2)
1, colored multi-form HCG test paper bar, it is characterized in that: this test strips is that mutual in turn overlap joint ground is pasted glass fibre membrane (3), coated film (4) and the thieving paper (5) of sample pad (2), coated with gold labeling antibody and formed on end liner (1), coated film (4) has detection line (6) and control line (7), detection line (6) bag is by HCG antibody, and control line (7) wraps by anti-mouse antibody; The colour developing color of described detection line (6) and control line (7) has diversity, is the size by the control colloid gold particle, obtains the colloid gold particle of different colours, perhaps adopts the emulsion particle of different colours; Described detection line (6) and control line
(7) colour developing form has diversity, is that spray is bar shaped, rhombus, circle, quincunx, the multiple difform band of cruciform by the setting program of Membrane jetter in the adjusting production technology.
2, the multi-form HCG test paper of colour according to claim 1 bar; it is characterized in that: the HCG antibody of described bag quilt is highly purified anti-monoclonal antibody and/or polyclonal antibody, the anti-β of mark-HCG monoclonal antibody on described colored colloid gold particle or the colored latex particle.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200520056329 CN2779402Y (en) | 2005-03-30 | 2005-03-30 | Chromatic multi-form test strip for HCG test |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200520056329 CN2779402Y (en) | 2005-03-30 | 2005-03-30 | Chromatic multi-form test strip for HCG test |
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| CN102192979A (en) * | 2010-03-08 | 2011-09-21 | 苏州浩欧博生物医药有限公司 | Lateral chromatographic one-step analytical method utilizing non-bibulous film, and assay kit |
| CN102192981A (en) * | 2010-03-10 | 2011-09-21 | 苏州浩欧博生物医药有限公司 | Direct-reading solid phase immune analysis method |
| CN102236014A (en) * | 2010-04-20 | 2011-11-09 | 艾博生物医药(杭州)有限公司 | Detection reagent strip based on immunoreaction principle |
| CN102272604A (en) * | 2008-11-17 | 2011-12-07 | 盖茨和圣托马斯英国国民健康制度保险信托基金会 | Pregnancy testing |
| CN102879576A (en) * | 2012-10-23 | 2013-01-16 | 广州万孚生物技术股份有限公司 | Early pregnancy detection test strip and detection method thereof |
| CN103558401A (en) * | 2013-11-04 | 2014-02-05 | 无锡博慧斯生物医药科技有限公司 | Human chorionic gonadotropin and bacterial vaginitis joint inspection kit |
| CN103777002A (en) * | 2014-01-15 | 2014-05-07 | 南通市伊士生物技术有限责任公司 | Preparation method of multifunctional test paper for early pregnancy |
| CN104471398A (en) * | 2012-11-28 | 2015-03-25 | 古河电气工业株式会社 | Immunochromatography, and detector and reagent for use therein |
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| CN113588958A (en) * | 2021-08-17 | 2021-11-02 | 合肥工业大学智能制造技术研究院 | Lamotrigine lateral chromatography test strip with multiple concentration detection lines and application thereof |
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| CN102272604A (en) * | 2008-11-17 | 2011-12-07 | 盖茨和圣托马斯英国国民健康制度保险信托基金会 | Pregnancy testing |
| CN102192979A (en) * | 2010-03-08 | 2011-09-21 | 苏州浩欧博生物医药有限公司 | Lateral chromatographic one-step analytical method utilizing non-bibulous film, and assay kit |
| CN102192981A (en) * | 2010-03-10 | 2011-09-21 | 苏州浩欧博生物医药有限公司 | Direct-reading solid phase immune analysis method |
| CN102236014A (en) * | 2010-04-20 | 2011-11-09 | 艾博生物医药(杭州)有限公司 | Detection reagent strip based on immunoreaction principle |
| CN102879576A (en) * | 2012-10-23 | 2013-01-16 | 广州万孚生物技术股份有限公司 | Early pregnancy detection test strip and detection method thereof |
| US10036750B2 (en) | 2012-11-28 | 2018-07-31 | Furukawa Electric Co., Ltd. | Immunochromatography, and detection device and reagent for the same |
| CN104471398A (en) * | 2012-11-28 | 2015-03-25 | 古河电气工业株式会社 | Immunochromatography, and detector and reagent for use therein |
| CN104471398B (en) * | 2012-11-28 | 2016-10-26 | 古河电气工业株式会社 | The detection device used in immunochromatographic method, the method |
| CN103558401A (en) * | 2013-11-04 | 2014-02-05 | 无锡博慧斯生物医药科技有限公司 | Human chorionic gonadotropin and bacterial vaginitis joint inspection kit |
| CN103777002A (en) * | 2014-01-15 | 2014-05-07 | 南通市伊士生物技术有限责任公司 | Preparation method of multifunctional test paper for early pregnancy |
| CN106290925A (en) * | 2016-07-31 | 2017-01-04 | 江翠珍 | Immunochromatography detection method and immunochromatographytest test kit |
| CN113588958A (en) * | 2021-08-17 | 2021-11-02 | 合肥工业大学智能制造技术研究院 | Lamotrigine lateral chromatography test strip with multiple concentration detection lines and application thereof |
| CN113588958B (en) * | 2021-08-17 | 2023-11-10 | 合肥工业大学智能制造技术研究院 | Lamotrigine lateral chromatography test strip with multiple concentration detection lines and application thereof |
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Effective date of registration: 20090206 Address after: 8, litchi mountain road, Luogang District Science Town, Guangdong City, Guangzhou Province, China: 510530 Patentee after: Wondfo Biotech Co., Ltd. Address before: South China University of Technology, Guangdong, Guangzhou Province, China, postcode: 510641 Patentee before: Wang Jihua |
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