CN103558401A - Human chorionic gonadotropin and bacterial vaginitis joint inspection kit - Google Patents
Human chorionic gonadotropin and bacterial vaginitis joint inspection kit Download PDFInfo
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- CN103558401A CN103558401A CN201310538905.XA CN201310538905A CN103558401A CN 103558401 A CN103558401 A CN 103558401A CN 201310538905 A CN201310538905 A CN 201310538905A CN 103558401 A CN103558401 A CN 103558401A
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
- G01N33/76—Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
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- G01N2333/59—Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
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Abstract
The invention relates to a human chorionic gonadotropin (HCG) and bacterial vaginitis (BV) joint inspection kit and belongs to the technical field of detection. The HCG and BV joint inspection kit comprises an HCG immunochromatographic assay test strip, and one or several in pH test paper, sialidase detection test paper, leukocyte esterase detection test paper, hydrogen peroxide detection test paper and amine test paper; preferably, a binding pad and a chromatography film of the immunochromatographic assay test strip are correspondingly coated with an immunolabeled antibody and a detection antibody respectively; the immunochromatographic assay test strip and one or several in the pH test paper, the sialidase detection test paper, the leukocyte esterase detection test paper, the hydrogen peroxide detection test paper and the amine test paper are arranged in parallel or arranged on the upper and lower surfaces of a bottom plate. According to the kit, the HCG and BV can be simultaneously detected through self-sampling, a joint inspection result provides strong evidence for prepotency, and the kit is simple, convenient and suitable for large-scale popularization and application.
Description
Technical field
The present invention relates to a kind of human chorionic gonadotrophin and bacterial vaginitis joint inspection kit, particularly a kind of human chorionic gonadotrophin (HCG) and bacterial vaginitis (BV) joint inspection kit, belong to detection technique field.
Background technology
Human chorionic gonadotrophin (HCG), it is a kind of glycoprotein hormones with promotion gonad development by the secretion of tire basin syneytiotrophoblast, be present in pregnant woman's blood, urine, colostrum, amniotic fluid and fetus body, in First Trimester HCG secretory volume, increase and be exceedingly fast, when all to pregnant 8-10, serum-concentration peaks, at the gestation initial stage, rule HCG ratio in serum is the highest, its content is substantially parallel in blood, urine, being determined at clinically of HCG correlation molecule is widely used, and comprising: for the diagnosis of early pregnancy; The appearance (as threatened abortion, Down syndrome etc.) of prompting abnormal pregnancy; Diagnose and monitor pregnant trophoblastic disease (vesicular mole, suede cancer etc.) etc.
Bacterial vaginitis (BV) refers to that a class shows as genital tract normal flora and (produces H on bacteriology
2o
2lactobacillus) quantity reduces, and replaces the clinical symptom grouping of one group of anaerobism group (Bacteroides family, Gardnerella, Mo Bilun Bordetella, mycoplasma hominis belong to and Peptostreptococcus etc.) quantity due to increasing.BV can lead to grave consequences to mother and fetus and baby's health, as: uterine cavity infects, and pelvic infection and fine hair amnion infect, PNI, and threatened abortion, premature labor, premature rupture of fetal membranes, the infant of low-birth weight etc., thereby its Accurate Diagnosis is had very important significance.Clinical standard is Amsel goldstandard: 1. vaginal fluid is emulsion form; 2. pH>4.5; 3. amine test is positive; 4. in vaginal fluid, detect clues cell (>20%).If 4. negative, but the visible a large amount of gram-negative coccobacilluses of smear Gram’s staining, nothing or accidental lactic acid bacteria also can be judged to the BV positive.Said method all has experience requirement to reviewer, and diagnostic criteria is difficult for grasping, and needs microscope smear, wastes time and energy, and is not easy to the clinical fast detecting of outpatient service.
Summary of the invention
The object of the invention is to overcome above-mentioned weak point, a kind of human chorionic gonadotrophin (HCG) and bacterial vaginitis (BV) joint inspection kit are provided, by oneself, sample, can fast detecting bacterial vaginitis, can accurately differentiate whether there is early pregnancy simultaneously.
According to technical scheme provided by the invention, and a bacterial vaginitis joint inspection kit, comprise the first base plate, in the one side of the first base plate, be provided with chromatographic film, the front end of chromatographic film arranges opaque diaphragm, and the front end of opaque diaphragm arranges arrow; The front end of arrow arranges pad; In chromatographic film, be also disposed with detection line and nature controlling line, chromatographic film rear end is provided with adsorptive pads.
Also comprise infection index detection zone.
Described infection index detection zone is attached on the second base plate or is compound on any one surface of the first base plate.
In the one side of the first base plate, be provided with chromatographic film, the front end of chromatographic film arranges pad, and the place ahead of pad also arranges sample pad; In chromatographic film, be also disposed with detection line and nature controlling line, chromatographic film rear end is provided with adsorptive pads; On the lower surface of the first base plate, be also provided with infection index detection zone.
Described infection index detection zone is provided with pH test paper, sialidase Test paper, leukocyte esterase Test paper, one or more in hydrogen peroxide Test paper and amine test paper.
The described pH test paper that adheres to, sialidase Test paper, leukocyte esterase Test paper, the first base plate of hydrogen peroxide Test paper and amine test paper or the material of the second base plate are a kind of in PET, PVC, PE, PP or PS, long is 10-300mm, wide 2-30mm, and thickness is 0.1-1.5mm.
Described pH test paper, sialidase Test paper, leukocyte esterase Test paper, the basic material of hydrogen peroxide Test paper and amine test paper is cellulose filter paper, glass fiber filter paper, chromatographic paper, polyester film or nylon membrane; Described test paper be shaped as square, rectangle, circle, ellipse, triangle, rhombus or trapezoidal; Size is long 1-30 mm; Wide 1-30 mm; Thickness 0.01-5 mm; Spacing between multiple test paper is 1-30 mm.
On described pad (1), be coated with anti-hCG HCG gold labelled antibody.
In described chromatographic film (6), be coated with anti-hCG HCG and detect antibody.
Beneficial effect of the present invention: the present invention can sample by oneself, can fast detecting bacterial vaginitis, can accurately differentiate whether there is early pregnancy simultaneously, simple and convenient, is applicable to large-scale promotion application.
Accompanying drawing explanation
Fig. 1 is embodiment 1 first base plate 10 structural representations.
Fig. 2 is embodiment 1 second base plate 11 structural representations.
Fig. 3 is embodiment 2 structural representations.
Fig. 4 is embodiment 3 structural representations.
Fig. 5 is embodiment 4 structural representations.
Fig. 6 is embodiment 1 infection index detection zone structural representation.
Fig. 7 is embodiment 2 infection index detection zone structural representations.
Fig. 8 is embodiment 3 infection index detection zone structural representations.
Fig. 9 is embodiment 4 infection index detection zone structural representations.
Embodiment
As shown in Fig. 1,2,5, and a bacterial vaginitis joint inspection kit, comprise the first base plate 10, in the one side of the first base plate 10, be provided with chromatographic film 6, the front end of chromatographic film 6 arranges opaque diaphragm 3, and the front end of opaque diaphragm 3 arranges arrow 2; The front end of arrow 2 arranges pad 1; In chromatographic film 6, be also disposed with detection line 4 and nature controlling line 5, chromatographic film 6 rear ends are provided with adsorptive pads 7.
Also comprise infection index detection zone 8.Described infection index detection zone 8 is attached to the upper surface of the first base plate 10.
Described infection index detection zone 8 is provided with described pH test paper.Described HCG immunochromatographydetecting detecting test strip, and urinary system infection contamination index detection zone, comprise described pH test paper, is placed in the upper surface of base plate 10 with horizontal coverage mode or built-in embedded mode.
As shown in Fig. 3,7, and a bacterial vaginitis joint inspection kit, comprise the first base plate 10, in the one side of the first base plate 10, be provided with chromatographic film 6, the front end of chromatographic film 6 arranges opaque diaphragm 3, and the front end of opaque diaphragm 3 arranges arrow 2; The front end of arrow 2 arranges pad 1; In chromatographic film 6, be also disposed with detection line 4 and nature controlling line 5, chromatographic film 6 rear ends are provided with adsorptive pads 7.
Also comprise infection index detection zone 8.Described infection index detection zone 8 is attached on the second base plate 11.
Described infection index detection zone 8 is provided with sialidase Test paper, leukocyte esterase Test paper.With horizontal coverage mode or built-in embedded mode, be placed in the upper surface of base plate, and be set up in parallel on draw-in groove with HCG immunochromatographydetecting detecting test strip.
As shown in Fig. 4,8, and a bacterial vaginitis joint inspection kit, comprise the first base plate 10, in the one side of the first base plate 10, be provided with chromatographic film 6, the front end of chromatographic film 6 arranges pad 1, and the place ahead of pad 1 also arranges sample pad 9; In chromatographic film 6, be also disposed with detection line 4 and nature controlling line 5, chromatographic film 6 rear ends are provided with adsorptive pads 7; On the lower surface of the first base plate 10, be also provided with infection index detection zone 8.
Described infection index detection zone 8 is attached on the lower surface of the first base plate 10.
Described infection index detection zone 8 is provided with sialidase Test paper, leukocyte esterase Test paper, hydrogen peroxide Test paper, is placed in the surface of base plate with horizontal coverage mode or built-in embedded mode, and is placed in respectively the upper and lower surface of base plate with HCG immunity chromatography detection test paper.
As shown in Fig. 5,9, and a bacterial vaginitis joint inspection kit, comprise the first base plate 10, in the one side of the first base plate 10, be provided with chromatographic film 6, the front end of chromatographic film 6 arranges opaque diaphragm 3, and the front end of opaque diaphragm 3 arranges arrow 2; The front end of arrow 2 arranges pad 1; In chromatographic film 6, be also disposed with detection line 4 and nature controlling line 5, chromatographic film 6 rear ends are provided with adsorptive pads 7.Described infection index detection zone 8 is attached on the second base plate 11.
In embodiment 1-4, the immune labeled antibody of described HCG is anti-human HCG colloid gold immune labelled antibody, it is that anti-human HCG detects antibody that described HCG detects antibody, and anti-human HCG colloid gold immune labelled antibody wherein and anti-human HCG detect antibody in conjunction with the different parts of people HCG;
Described infection index detection zone 8 is provided with sialidase Test paper, leukocyte esterase Test paper, hydrogen peroxide Test paper and amine test paper, be placed in the upper surface of base plate, and be placed on dismountable draw-in groove with HCG immunity chromatography detection test paper with horizontal coverage mode or built-in embedded mode.
Described pH test paper is dry processing after blank filter paper is soaked in the solution of methyl orange and bromcresol green composition, and the concentration of described methyl orange in solution is 10-600 mg/L, and the concentration of described bromcresol green in solution is 10-300 mg/L;
Described sialidase Test paper is for to be immersed in the chloro-3-indoles of the bromo-4-of 5-acetyl neuraminic acid salt by blank filter paper, NBT, dry processing after soaking in the solution that trehalose and PBS damping fluid form, the concentration of the chloro-3-indoles of the bromo-4-of described 5-acetyl neuraminic acid salt in solution is 5-200 mg/L, the concentration of described NBT in solution is 10-500 mg/L, described trehalose is 200-10000 mg/L in the concentration in solution, the pH value of described PBS is 6.5, and volumetric molar concentration is 0.01-1 M;
Described leukocyte esterase Test paper is dry processing after blank filter paper is soaked in the solution of the chloro-3-indolyl acetic acid salt of the bromo-4-of 5-and sucrose composition, the concentration of the chloro-3-indolyl acetic acid salt of the bromo-4-of described 5-in solution is 40-1000 mg/L, and the concentration of described sucrose in solution is 200-10000 mg/L;
Described hydrogen peroxide Test paper be by blank filter paper at peroxidase, dry processing after soaking in the enzyme solutions that surfactant and tetramethyl benzidine (TMB) form, the concentration of described peroxidase in solution is 1.0-3.0 * 10
5u/L, the concentration of surfactant in solution is 0.1-10%, the concentration of TMB in enzyme solutions is 200-1000mg/L;
The dry reagent piece B processing after described amine test paper soaks in bromcresol green and Triton X-100 solution for the dry reagent piece A processing after blank filter paper is soaked in potassium hydroxide solution is pasted on, wherein in A reagent piece, concentration of potassium hydroxide is 400-10000 mg/L, in B reagent piece, bromcresol green concentration is 100-1000, and Triton X-100 is 0.1-10 %.
Sample dilution is 0.9 % NaCl.
Application Example 1
Use the operation steps of inventor's human chorionic gonadtropin (HCG) and bacterial vaginitis (BV) joint inspection kit as follows:
(1) with cotton swab, from posterior fornix, getting secretion is first directly coated in pH test paper;
(2) with cotton swab, from posterior fornix, get secretion again, add the sample diluted secretion of 300 ~ 500 μ L;
(3) respectively at HCG immunochromatographydetecting detecting test strip, sialidase Test paper, leukocyte esterase Test paper, drips the secretion of appropriate dilution on hydrogen peroxide Test paper and amine test paper.
(4) by test strips at room temperature standing about 10 minutes or in 37 ℃ of constant temperature ovens visual colorimetric determination after standing about 10 minutes.
Sentence read result: if HCG immunochromatographydetecting detecting test strip C district occurs that band ，Ze T district occurs that band represents that sample is positive, does not occur that band represents feminine gender during detection; If band does not appear in C district, represent that test strips lost efficacy.PH test paper occurs red or light green expression is normal, occurs that bottle green represents undesired.Sialidase Test paper does not develop the color and represents normally, to occur that purple or brown represent undesired.Leukocyte esterase Test paper does not develop the color and represents normally, occurs that blue or green represents undesired.There is blue expression normally in hydrogen peroxide Test paper, not developing the color, it is undesired to represent.Amine test paper does not develop the color and represents normally, occurs that green expression is undesired.
Preparation Example 1
The antibody colloidal gold labeling method detecting for HCG is for adding 400 μ L, and 3% sal tartari regulates the pH of 50 mL colloidal gold solutions; Under stirring condition, add gradually 0.25 mL, 6.6 mg/mL anti-HCG antibody to be marked, in above-mentioned colloidal gold solution, mixes standing 30 min of room temperature; 12.5 mL confining liquids are slowly added in above-mentioned colloidal gold solution, mix, standing 30 min of room temperature; The centrifugal 1h of 8000 4 ℃ of g, removes supernatant, stays loose deposits; Each pipe precipitation is dissolved with 25 mL cleansing solutions, and the centrifugal 1h of 6,000 4 ℃ of g, removes supernatant, stays loose deposits; Each pipe precipitation is dissolved with 1.2 mL cleansing solutions, proceeds in 1.5 mL centrifuge tubes, and the centrifugal 1h of 4,500 4 ℃ of g, removes supernatant; All precipitations are resuspended with the resuspended liquid of 1 mL colloidal gold antibody, 4 ℃ of preservations.
The specking method of the antibody colloidal gold pad detecting for HCG is as follows: with the resuspended liquid of colloidal gold antibody, the colloidal gold antibody bond of above-mentioned preparation is diluted to 4 times; The power supply of opening point film instrument, sets specking program, and specking amount is 8 μ L/cm; No. 1 pipeline is specking passage; No. 1 pipeline is placed in to the resuspended solution of colloidal gold antibody, selects initialize routine, 6 circulations of initialization; Gold mark pad is lain on a film instrument by fixed position, press " GO " key on control panel and start specking, after having put, take off, check the good gold mark pad of specking, the colloidal gold antibody band of specking evenly, continuously and the straight line that connects whole gold mark pad be qualified specking product, in two straight lines, occur that breakpoint is defective specking product; Often put a slice gold mark pad, " GO " key of pressing on a control panel is specking once (a slice); Specking finishes, and the gold mark pad of specking is placed in to room temperature natural drying 1 hour, on film, should can't see specking vestige.
The antibody chromatography membrane preparation method detecting for HCG is as follows: get mouse-anti people HCG antibody 500 ug, be added in 5 mL graduated centrifuge tubes, antibody diluent to 1 mL, Container Tag T sign.Get sheep anti-mouse igg antibody 25 μ L, be added in 5 mL graduated centrifuge tubes, antibody diluent to 1 mL, Container Tag C sign; The power supply of opening point film instrument, sets specking program, and specking amount is 1 μ L/cm, and No. 1 pipeline is for detecting band specking passage, and No. 2 pipelines are contrast band specking passage; No. 1 pipeline is placed in and detects band solution, No. 2 pipelines are placed in to contrast band solution, select initialize routine, 6 circulations of initialization; Chromatographic film is lain on a film instrument by fixed position, press " GO " key on control panel and start specking, after having put, take off, check the good chromatographic film of specking, detect band and contrast band and be two evenly, continuously and the straight line that connects whole chromatographic film be qualified specking product, in two straight lines, occur that breakpoint is defective specking product; Often put a slice chromatographic film, " GO " key of pressing on a control panel is specking once (a slice); Specking finishes, and the chromatographic film of specking is placed in to room temperature natural drying 1 hour, on film, should can't see specking vestige.
The protection sheet of wider portion on base plate is removed, and edge is the lower limb of protection sheet above, will pull the chromatographic film of line, in C line mode up, is attached on base plate plate; Collaurum pad is attached to T line below, and a little contacts with NC film; Sample pad is attached to collaurum pad below, and a little contacts with collaurum pad; Then remove top protection sheet, adsorptive pads is attached to the top of NC film, a little contacts with NC film; Protection sheet and index strip paper are attached to the test strips outside assembling one by one, are assembled into kilocalorie.Connect cutting machine power supply, set the film program of cutting, setting cutting width is 4 mm; Kilocalorie certified products are kept flat in cutting machine platform track, face up, press " GO " key on guidance panel, start cutting; Often put a slice kilocalorie certified products, press on guidance panel " GO " key once, until cut all kilocalorie certified products.
PH test paper, sialidase Test paper, leukocyte esterase Test paper, the filter paper material of hydrogen peroxide Test paper and amine test paper is chromatographic paper and nylon membrane, the material that base plate is used is transparent PVC sheet.
PH test paper is made: it is 300 mg/L that filter paper is immersed in to methyl orange concentration, and bromcresol green concentration is in the solution of 200 mg/L, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Sialidase Test paper is made: it is 200 mg/L that filter paper is immersed in to the chloro-3-indoles of the bromo-4-of 5-acetyl neuraminic acid salinity, NBT concentration is 400 mg/L, trehalose concentration is 2000 mg/L, the pH value of PBS damping fluid is 6.5, volumetric molar concentration is in the solution of 0.01 M, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Leukocyte esterase Test paper is made: it is 300 mg/L that filter paper is immersed in to the chloro-3-indolyl acetic acid salinity of the bromo-4-of 5-, and sucrose concentration is in the solution of 2000 mg/L, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Hydrogen peroxide Test paper is made: it is 2.0 * 10 that filter paper is immersed in to peroxidase concn
5u/L, the concentration of surfactant in solution is 0.2 %, the concentration of TMB in enzyme solutions is in the solution of 700 mg/L, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Amine test paper is made: reagent piece A, for filter paper is immersed in the solution that concentration of potassium hydroxide is 10000 mg/L, is dried; Reagent piece B is 500 mg/L for nylon membrane is immersed in to bromcresol green concentration, in the solution that Triton X-100 is 0.2%, dries.By reagent piece A and B with being cut into cutting machine the square that the length of side is 4 mm after adhered by double sided plaster;
Respectively by pH test paper, sialidase Test paper, leukocyte esterase Test paper, one or more in hydrogen peroxide Test paper and amine test paper cover with level or the stickup of built-in embedded mode is fixed on transparent PVC sheet, each test paper spacing is 5 mm, is cut into the length of side and is the test strips of 4 mm.
Preparation Example 2
The antibody colloidal gold labeling method detecting for HCG is for adding 400 μ L, and 3% sal tartari regulates the pH of 50 mL colloidal gold solutions; Under stirring condition, add gradually 0.25 mL, 6.6 mg/mL anti-HCG antibody to be marked, in above-mentioned colloidal gold solution, mixes standing 30 min of room temperature; 12.5 mL confining liquids are slowly added in above-mentioned colloidal gold solution, mix, standing 30 min of room temperature; The centrifugal 1h of 8000 4 ℃ of g, removes supernatant, stays loose deposits; Each pipe precipitation is dissolved with 25 mL cleansing solutions, and the centrifugal 1h of 6,000 4 ℃ of g, removes supernatant, stays loose deposits; Each pipe precipitation is dissolved with 1.2 mL cleansing solutions, proceeds in 1.5 mL centrifuge tubes, and the centrifugal 1h of 4,500 4 ℃ of g, removes supernatant; All precipitations are resuspended with the resuspended liquid of 1 mL colloidal gold antibody, 4 ℃ of preservations.
The specking method of the antibody colloidal gold pad detecting for HCG is as follows: with the resuspended liquid of colloidal gold antibody, the colloidal gold antibody bond of above-mentioned preparation is diluted to 4 times; The power supply of opening point film instrument, sets specking program, and specking amount is 8 μ L/cm; No. 1 pipeline is specking passage; No. 1 pipeline is placed in to the resuspended solution of colloidal gold antibody, selects initialize routine, 6 circulations of initialization; Gold mark pad is lain on a film instrument by fixed position, press " GO " key on control panel and start specking, after having put, take off, check the good gold mark pad of specking, the colloidal gold antibody band of specking evenly, continuously and the straight line that connects whole gold mark pad be qualified specking product, in two straight lines, occur that breakpoint is defective specking product; Often put a slice gold mark pad, " GO " key of pressing on a control panel is specking once (a slice); Specking finishes, and the gold mark pad of specking is placed in to room temperature natural drying 1 hour, on film, should can't see specking vestige.
The antibody chromatography membrane preparation method detecting for HCG is as follows: get mouse-anti people HCG antibody 500 ug, be added in 5 mL graduated centrifuge tubes, antibody diluent to 1 mL, Container Tag T sign.Get sheep anti-mouse igg antibody 25 μ L, be added in 5 mL graduated centrifuge tubes, antibody diluent to 1 mL, Container Tag C sign; The power supply of opening point film instrument, sets specking program, and specking amount is 1 μ L/cm, and No. 1 pipeline is for detecting band specking passage, and No. 2 pipelines are contrast band specking passage; No. 1 pipeline is placed in and detects band solution, No. 2 pipelines are placed in to contrast band solution, select initialize routine, 6 circulations of initialization; Chromatographic film is lain on a film instrument by fixed position, press " GO " key on control panel and start specking, after having put, take off, check the good chromatographic film of specking, detect band and contrast band and be two evenly, continuously and the straight line that connects whole chromatographic film be qualified specking product, in two straight lines, occur that breakpoint is defective specking product; Often put a slice chromatographic film, " GO " key of pressing on a control panel is specking once (a slice); Specking finishes, and the chromatographic film of specking is placed in to room temperature natural drying 1 hour, on film, should can't see specking vestige.
The protection sheet of wider portion on base plate is removed, and edge is the lower limb of protection sheet above, will pull the chromatographic film of line, in C line mode up, is attached on base plate plate; Collaurum pad is attached to T line below, and a little contacts with NC film; Sample pad is attached to collaurum pad below, and a little contacts with collaurum pad; Then remove top protection sheet, adsorptive pads is attached to the top of NC film, a little contacts with NC film; Protection sheet and index strip paper are attached to the test strips outside assembling one by one, are assembled into kilocalorie.Connect cutting machine power supply, set the film program of cutting, setting cutting width is 4 mm; Kilocalorie certified products are kept flat in cutting machine platform track, face up, press " GO " key on guidance panel, start cutting; Often put a slice kilocalorie certified products, press on guidance panel " GO " key once, until cut all kilocalorie certified products.
PH test paper, sialidase Test paper, leukocyte esterase Test paper, the filter paper material of hydrogen peroxide Test paper and amine test paper is chromatographic paper and nylon membrane, the material that base plate is used is transparent PVC sheet.
PH test paper is made: it is 300 mg/L that filter paper is immersed in to methyl orange concentration, and bromcresol green concentration is in the solution of 200 mg/L, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Sialidase Test paper is made: it is 200 mg/L that filter paper is immersed in to the chloro-3-indoles of the bromo-4-of 5-acetyl neuraminic acid salinity, NBT concentration is 400 mg/L, trehalose concentration is 2000 mg/L, the pH value of PBS damping fluid is 6.5, volumetric molar concentration is in the solution of 0.01 M, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Leukocyte esterase Test paper is made: it is 300 mg/L that filter paper is immersed in to the chloro-3-indolyl acetic acid salinity of the bromo-4-of 5-, and sucrose concentration is in the solution of 2000 mg/L, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Hydrogen peroxide Test paper is made: it is 2.0 * 10 that filter paper is immersed in to peroxidase concn
5u/L, the concentration of surfactant in solution is 0.2 %, the concentration of TMB in enzyme solutions is in the solution of 700 mg/L, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Amine test paper is made: reagent piece A, for filter paper is immersed in the solution that concentration of potassium hydroxide is 10000 mg/L, is dried; Reagent piece B is 500 mg/L for nylon membrane is immersed in to bromcresol green concentration, in the solution that Triton X-100 is 0.2%, dries.By reagent piece A and B with being cut into cutting machine the square that the length of side is 4 mm after adhered by double sided plaster;
By the pH test paper of well cutting, sialidase Test paper, leukocyte esterase Test paper, one or more in hydrogen peroxide Test paper and amine test paper cover with level or the mode of built-in embedding is placed on transparent PVC base plate, and are set up in parallel on draw-in groove with HCG immunochromatographydetecting detecting test strip.
Preparation Example 3
The antibody colloidal gold labeling method detecting for HCG is for adding 400 μ L, and 3% sal tartari regulates the pH of 50 mL colloidal gold solutions; Under stirring condition, add gradually 0.25 mL, 6.6 mg/mL anti-HCG antibody to be marked, in above-mentioned colloidal gold solution, mixes standing 30 min of room temperature; 12.5 mL confining liquids are slowly added in above-mentioned colloidal gold solution, mix, standing 30 min of room temperature; The centrifugal 1h of 8000 4 ℃ of g, removes supernatant, stays loose deposits; Each pipe precipitation is dissolved with 25 mL cleansing solutions, and the centrifugal 1h of 6,000 4 ℃ of g, removes supernatant, stays loose deposits; Each pipe precipitation is dissolved with 1.2 mL cleansing solutions, proceeds in 1.5 mL centrifuge tubes, and the centrifugal 1h of 4,500 4 ℃ of g, removes supernatant; All precipitations are resuspended with the resuspended liquid of 1 mL colloidal gold antibody, 4 ℃ of preservations.
The specking method of the antibody colloidal gold pad detecting for HCG is as follows: with the resuspended liquid of colloidal gold antibody, the colloidal gold antibody bond of above-mentioned preparation is diluted to 4 times; The power supply of opening point film instrument, sets specking program, and specking amount is 8 μ L/cm; No. 1 pipeline is specking passage; No. 1 pipeline is placed in to the resuspended solution of colloidal gold antibody, selects initialize routine, 6 circulations of initialization; Gold mark pad is lain on a film instrument by fixed position, press " GO " key on control panel and start specking, after having put, take off, check the good gold mark pad of specking, the colloidal gold antibody band of specking evenly, continuously and the straight line that connects whole gold mark pad be qualified specking product, in two straight lines, occur that breakpoint is defective specking product; Often put a slice gold mark pad, " GO " key of pressing on a control panel is specking once (a slice); Specking finishes, and the gold mark pad of specking is placed in to room temperature natural drying 1 hour, on film, should can't see specking vestige.
The antibody chromatography membrane preparation method detecting for HCG is as follows: get mouse-anti people HCG antibody 500 ug, be added in 5 mL graduated centrifuge tubes, antibody diluent to 1 mL, Container Tag T sign.Get sheep anti-mouse igg antibody 25 μ L, be added in 5 mL graduated centrifuge tubes, antibody diluent to 1 mL, Container Tag C sign; The power supply of opening point film instrument, sets specking program, and specking amount is 1 μ L/cm, and No. 1 pipeline is for detecting band specking passage, and No. 2 pipelines are contrast band specking passage; No. 1 pipeline is placed in and detects band solution, No. 2 pipelines are placed in to contrast band solution, select initialize routine, 6 circulations of initialization; Chromatographic film is lain on a film instrument by fixed position, press " GO " key on control panel and start specking, after having put, take off, check the good chromatographic film of specking, detect band and contrast band and be two evenly, continuously and the straight line that connects whole chromatographic film be qualified specking product, in two straight lines, occur that breakpoint is defective specking product; Often put a slice chromatographic film, " GO " key of pressing on a control panel is specking once (a slice); Specking finishes, and the chromatographic film of specking is placed in to room temperature natural drying 1 hour, on film, should can't see specking vestige.
The protection sheet of wider portion on base plate is removed, and edge is the lower limb of protection sheet above, will pull the chromatographic film of line, in C line mode up, is attached on base plate plate; Collaurum pad is attached to T line below, and a little contacts with NC film; Sample pad is attached to collaurum pad below, and a little contacts with collaurum pad; Then remove top protection sheet, adsorptive pads is attached to the top of NC film, a little contacts with NC film; Protection sheet and index strip paper are attached to the test strips outside assembling one by one, are assembled into kilocalorie.Connect cutting machine power supply, set the film program of cutting, setting cutting width is 4 mm; Kilocalorie certified products are kept flat in cutting machine platform track, face up, press " GO " key on guidance panel, start cutting; Often put a slice kilocalorie certified products, press on guidance panel " GO " key once, until cut all kilocalorie certified products.
PH test paper, sialidase Test paper, leukocyte esterase Test paper, the filter paper material of hydrogen peroxide Test paper and amine test paper is chromatographic paper and nylon membrane, the material that base plate is used is transparent PVC sheet.
PH test paper is made: it is 300 mg/L that filter paper is immersed in to methyl orange concentration, and bromcresol green concentration is in the solution of 200 mg/L, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Sialidase Test paper is made: it is 200 mg/L that filter paper is immersed in to the chloro-3-indoles of the bromo-4-of 5-acetyl neuraminic acid salinity, NBT concentration is 400 mg/L, trehalose concentration is 2000 mg/L, the pH value of PBS damping fluid is 6.5, volumetric molar concentration is in the solution of 0.01 M, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Leukocyte esterase Test paper is made: it is 300 mg/L that filter paper is immersed in to the chloro-3-indolyl acetic acid salinity of the bromo-4-of 5-, and sucrose concentration is in the solution of 2000 mg/L, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Hydrogen peroxide Test paper is made: it is 2.0 * 10 that filter paper is immersed in to peroxidase concn
5u/L, the concentration of surfactant in solution is 0.2 %, the concentration of TMB in enzyme solutions is in the solution of 700 mg/L, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Amine test paper is made: reagent piece A, for filter paper is immersed in the solution that concentration of potassium hydroxide is 10000 mg/L, is dried; Reagent piece B is 500 mg/L for nylon membrane is immersed in to bromcresol green concentration, in the solution that Triton X-100 is 0.2%, dries.By reagent piece A and B with being cut into cutting machine the square that the length of side is 4 mm after adhered by double sided plaster;
By the pH test paper of well cutting, sialidase Test paper, leukocyte esterase Test paper, one or more in hydrogen peroxide Test paper and amine test paper cover with level or the mode of built-in embedding is placed on transparent PVC base plate, and are placed in respectively the upper and lower surface of base plate with HCG immunochromatographydetecting detecting test strip.
Preparation Example 4
The antibody colloidal gold labeling method detecting for HCG is for adding 400 μ L, and 3% sal tartari regulates the pH of 50 mL colloidal gold solutions; Under stirring condition, add gradually 0.25 mL, 6.6 mg/mL anti-HCG antibody to be marked, in above-mentioned colloidal gold solution, mixes standing 30 min of room temperature; 12.5 mL confining liquids are slowly added in above-mentioned colloidal gold solution, mix, standing 30 min of room temperature; The centrifugal 1h of 8000 4 ℃ of g, removes supernatant, stays loose deposits; Each pipe precipitation is dissolved with 25 mL cleansing solutions, and the centrifugal 1h of 6,000 4 ℃ of g, removes supernatant, stays loose deposits; Each pipe precipitation is dissolved with 1.2 mL cleansing solutions, proceeds in 1.5 mL centrifuge tubes, and the centrifugal 1h of 4,500 4 ℃ of g, removes supernatant; All precipitations are resuspended with the resuspended liquid of 1 mL colloidal gold antibody, 4 ℃ of preservations.
The specking method of the antibody colloidal gold pad detecting for HCG is as follows: with the resuspended liquid of colloidal gold antibody, the colloidal gold antibody bond of above-mentioned preparation is diluted to 4 times; The power supply of opening point film instrument, sets specking program, and specking amount is 8 μ L/cm; No. 1 pipeline is specking passage; No. 1 pipeline is placed in to the resuspended solution of colloidal gold antibody, selects initialize routine, 6 circulations of initialization; Gold mark pad is lain on a film instrument by fixed position, press " GO " key on control panel and start specking, after having put, take off, check the good gold mark pad of specking, the colloidal gold antibody band of specking evenly, continuously and the straight line that connects whole gold mark pad be qualified specking product, in two straight lines, occur that breakpoint is defective specking product; Often put a slice gold mark pad, " GO " key of pressing on a control panel is specking once (a slice); Specking finishes, and the gold mark pad of specking is placed in to room temperature natural drying 1 hour, on film, should can't see specking vestige.
The antibody chromatography membrane preparation method detecting for HCG is as follows: get mouse-anti people HCG antibody 500 ug, be added in 5 mL graduated centrifuge tubes, antibody diluent to 1 mL, Container Tag T sign.Get sheep anti-mouse igg antibody 25 μ L, be added in 5 mL graduated centrifuge tubes, antibody diluent to 1 mL, Container Tag C sign; The power supply of opening point film instrument, sets specking program, and specking amount is 1 μ L/cm, and No. 1 pipeline is for detecting band specking passage, and No. 2 pipelines are contrast band specking passage; No. 1 pipeline is placed in and detects band solution, No. 2 pipelines are placed in to contrast band solution, select initialize routine, 6 circulations of initialization; Chromatographic film is lain on a film instrument by fixed position, press " GO " key on control panel and start specking, after having put, take off, check the good chromatographic film of specking, detect band and contrast band and be two evenly, continuously and the straight line that connects whole chromatographic film be qualified specking product, in two straight lines, occur that breakpoint is defective specking product; Often put a slice chromatographic film, " GO " key of pressing on a control panel is specking once (a slice); Specking finishes, and the chromatographic film of specking is placed in to room temperature natural drying 1 hour, on film, should can't see specking vestige.
The protection sheet of wider portion on base plate is removed, and edge is the lower limb of protection sheet above, will pull the chromatographic film of line, in C line mode up, is attached on base plate plate; Collaurum pad is attached to T line below, and a little contacts with NC film; Sample pad is attached to collaurum pad below, and a little contacts with collaurum pad; Then remove top protection sheet, adsorptive pads is attached to the top of NC film, a little contacts with NC film; Protection sheet and index strip paper are attached to the test strips outside assembling one by one, are assembled into kilocalorie.Connect cutting machine power supply, set the film program of cutting, setting cutting width is 4 mm; Kilocalorie certified products are kept flat in cutting machine platform track, face up, press " GO " key on guidance panel, start cutting; Often put a slice kilocalorie certified products, press on guidance panel " GO " key once, until cut all kilocalorie certified products.
PH test paper, sialidase Test paper, leukocyte esterase Test paper, the filter paper material of hydrogen peroxide Test paper and amine test paper is chromatographic paper and nylon membrane, the material that base plate is used is transparent PVC sheet.
PH test paper is made: it is 300 mg/L that filter paper is immersed in to methyl orange concentration, and bromcresol green concentration is in the solution of 200 mg/L, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Sialidase Test paper is made: it is 200 mg/L that filter paper is immersed in to the chloro-3-indoles of the bromo-4-of 5-acetyl neuraminic acid salinity, NBT concentration is 400 mg/L, trehalose concentration is 2000 mg/L, the pH value of PBS damping fluid is 6.5, volumetric molar concentration is in the solution of 0.01 M, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Leukocyte esterase Test paper is made: it is 300 mg/L that filter paper is immersed in to the chloro-3-indolyl acetic acid salinity of the bromo-4-of 5-, and sucrose concentration is in the solution of 2000 mg/L, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Hydrogen peroxide Test paper is made: it is 2.0 * 10 that filter paper is immersed in to peroxidase concn
5u/L, the concentration of surfactant in solution is 0.2 %, the concentration of TMB in enzyme solutions is in the solution of 700 mg/L, after drying, uses cutting machine to be cut into the square that the length of side is 4 mm;
Amine test paper is made: reagent piece A, for filter paper is immersed in the solution that concentration of potassium hydroxide is 10000 mg/L, is dried; Reagent piece B is 500 mg/L for nylon membrane is immersed in to bromcresol green concentration, in the solution that Triton X-100 is 0.2%, dries.By reagent piece A and B with being cut into cutting machine the square that the length of side is 4 mm after adhered by double sided plaster;
By the pH test paper of well cutting, sialidase Test paper, leukocyte esterase Test paper, one or more in hydrogen peroxide Test paper and amine test paper cover with level or the mode of built-in embedding is placed on transparent PVC base plate, and are arranged on dismountable draw-in groove with HCG immunochromatographydetecting detecting test strip.
Claims (9)
1. a human chorionic gonadotrophin and bacterial vaginitis joint inspection kit, comprise the first base plate (10), it is characterized in that: in the one side of the first base plate (10), be provided with chromatographic film (6), the front end of chromatographic film (6) arranges opaque diaphragm (3), and the front end of opaque diaphragm (3) arranges arrow (2); The front end of arrow (2) arranges pad (1); In chromatographic film (6), be also disposed with detection line (4) and nature controlling line (5), chromatographic film (6) rear end is provided with adsorptive pads (7).
2. human chorionic gonadotrophin and bacterial vaginitis joint inspection kit as claimed in claim 1, is characterized in that: also comprise infection index detection zone (8).
3. human chorionic gonadotrophin and bacterial vaginitis joint inspection kit as claimed in claim 2, is characterized in that: it is upper or be compound on any one surface of the first base plate (10) that described infection index detection zone (8) is attached to the second base plate (11).
4. a human chorionic gonadotrophin and bacterial vaginitis joint inspection kit, comprise the first base plate (10), it is characterized in that: in the one side of the first base plate (10), be provided with chromatographic film (6), the front end of chromatographic film (6) arranges pad (1), and the place ahead of pad (1) also arranges sample pad (9); In chromatographic film (6), be also disposed with detection line (4) and nature controlling line (5), chromatographic film (6) rear end is provided with adsorptive pads (7); On the lower surface of the first base plate (10), be also provided with infection index detection zone (8).
5. human chorionic gonadotrophin and bacterial vaginitis joint inspection kit as described in claim 2 or 4, it is characterized in that: described infection index detection zone (8) is provided with pH test paper, sialidase Test paper, leukocyte esterase Test paper, one or more in hydrogen peroxide Test paper and amine test paper.
6. human chorionic gonadotrophin and bacterial vaginitis joint inspection kit as claimed in claim 5, it is characterized in that: described in adhere to pH test paper, sialidase Test paper, leukocyte esterase Test paper, first base plate (10) of hydrogen peroxide Test paper and amine test paper or the material of the second base plate (11) are a kind of in PET, PVC, PE, PP or PS, long is 10-300mm, wide 2-30mm, and thickness is 0.1-1.5mm.
7. human chorionic gonadotrophin and bacterial vaginitis joint inspection kit as claimed in claim 5, it is characterized in that: described pH test paper, sialidase Test paper, leukocyte esterase Test paper, the basic material of hydrogen peroxide Test paper and amine test paper is cellulose filter paper, glass fiber filter paper, chromatographic paper, polyester film or nylon membrane; Described test paper be shaped as square, rectangle, circle, ellipse, triangle, rhombus or trapezoidal; Size is long 1-30 mm; Wide 1-30 mm; Thickness 0.01-5 mm; Spacing between multiple test paper is 1-30 mm.
8. human chorionic gonadotrophin and bacterial vaginitis joint inspection kit as described in claim 1 or 4, is characterized in that: on described pad (1), be coated with anti-hCG HCG gold labelled antibody.
9. human chorionic gonadotrophin and bacterial vaginitis joint inspection kit as described in claim 1 or 4, is characterized in that: in described chromatographic film (6), be coated with anti-hCG HCG and detect antibody.
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| CN201310538905.XA CN103558401A (en) | 2013-11-04 | 2013-11-04 | Human chorionic gonadotropin and bacterial vaginitis joint inspection kit |
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| CN201310538905.XA CN103558401A (en) | 2013-11-04 | 2013-11-04 | Human chorionic gonadotropin and bacterial vaginitis joint inspection kit |
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| CN108330159A (en) * | 2018-03-15 | 2018-07-27 | 深圳市瀚德标检生物工程有限公司 | A kind of detection reagent card and kit |
| CN117929722A (en) * | 2024-03-21 | 2024-04-26 | 山东康华生物医疗科技股份有限公司 | Multi-item combined detection kit for drug abuse in urine sample and saliva sample |
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| CN117929722A (en) * | 2024-03-21 | 2024-04-26 | 山东康华生物医疗科技股份有限公司 | Multi-item combined detection kit for drug abuse in urine sample and saliva sample |
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Application publication date: 20140205 |