CN203385727U - Enzyme-linked immunospot joint kit for family-planning four items - Google Patents
Enzyme-linked immunospot joint kit for family-planning four items Download PDFInfo
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- CN203385727U CN203385727U CN201320512939.7U CN201320512939U CN203385727U CN 203385727 U CN203385727 U CN 203385727U CN 201320512939 U CN201320512939 U CN 201320512939U CN 203385727 U CN203385727 U CN 203385727U
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Abstract
The utility model relates to an enzyme-linked immunospot joint kit for family-planning four items. The joint kit comprises a carrier plate, detecting lines/points for cytomegalovirus antibody/antigen, toxoplasma gondii antibody/antigen, rubella virus antibody/antigen, and herpes simplex virus antibody/antigen, a control line/point, a nitrocellulose membrane and the like, wherein the nitrocellulose membrane is adhered to the surface of the carrier plate, the detecting points for the cytomegalovirus antibody/antigen, the toxoplasma gondii antibody/antigen, the rubella virus antibody/antigen, and the herpes simplex virus antibody/antigen and the control point are sequentially arranged on the nitrocellulose membrane from the top to bottom, the detecting points for the cytomegalovirus antibody/antigen, the toxoplasma gondii antibody/antigen, the rubella virus antibody/antigen, and the herpes simplex virus antibody/antigen are enveloped by the corresponding antigen/antibody respectively, and the control line/point is enveloped by the cytomegalovirus antibody/antigen, the toxoplasma gondii antibody/antigen, the rubella virus antibody/antigen, and the herpes simplex virus antibody/antigen or patient positive serum containing the cytomegalovirus antibody/antigen, the toxoplasma gondii antibody/antigen, the rubella virus antibody/antigen, and the herpes simplex virus antibody/antigen. The joint kit provided by the utility model can be applied to the fast and one-time detection of the family-planning four items, and is simple to operate, low in cost, novel in design, and easy to observe and make a diagnosis.
Description
Technical field:
The utility model relates to the kit of a kind of elispot assay four kinds of viruses of disposable detection family planning (kind), belongs to four associating kit detection technique fields of medical diagnosis family planning.
Background technology:
It is an important detection method that is intended to prevent Newborn Birth-defects of carrying out at early pregnancy for the women of child-bearing age that four family plannings detect, it is for improving the overall quality of newborns, reduce neonate's risk of being born and have great importance, by medical institutions and pregnant and lying-in women, accepted extensively and apply.Four family plannings detect and comprise rubella virus, cytomegalovirus, toxoplasm and herpes simplex virus.Detect and can detect the situation whether pregnant woman and fetus exist infection by four family plannings, and treatment or terminal pregnancy targetedly.
When at present the application immune spot-ing detects this four diseases of diagnosis, the detection of every disease operate separately, and various reagent is and disperses placement, there is no special spot reaction plate, so testing process is complicated, time length, carry and inconvenient operation.As: chemical reaction method and Elisa method are all once a kind of operates and uses respectively kit separately.
Summary of the invention:
The purpose of this utility model is to overcome the shortcoming of prior art, be four of family plannings that four kinds of different disease kinds are put on a reaction plate, disposable operation can draw the diagnostic result of four kinds of diseases, reduced running program frequently, save human and material resources, save time, be easy to observe and make diagnosis.
In order to realize the foregoing invention purpose, the utility model is provided with carrier board, cytomegalovirus (CMV) antibody (antigen) detection line (point), Infection of Toxoplasma Gondii (TO) antibody (antigen) detection line (point), rubella virus (RV) antibody (antigen) detection line (point), herpes simplex virus (HSV-1) antibody (antigen) detection line (point), control line (point), nitrocellulose filter (NC film), incubation groove; Nitrocellulose filter sticks on the surface of carrier board, CMV antibody (antigen) detection line (point), toxoplasma antibody (antigen) detection line (point), Detecting Rubella Virus Antibodies In Human Sera (antigen) detection line (point), herpes simplex virus antibody (antigen) detection line (point), control line (point) are located on nitrocellulose filter from top to bottom successively, and are shaped with the incubation groove at each detection line (point) and control line (point) surrounding; Locate to be coated with respectively corresponding antigen (antibody) at CMV antibody (antigen) detection line (point), toxoplasma antibody (antigen) detection line (point), Detecting Rubella Virus Antibodies In Human Sera (antigen) detection line (point), herpes simplex virus antibody (antigen) detection line (point); Be coated with respectively from left to right cytomegalovirus antigen, toxoplasma antigen, rubella virus antigen, herpes simplex virus antigens or be coated with respectively the patient's positive serum containing CMV antibody, toxoplasma antibody, Detecting Rubella Virus Antibodies In Human Sera, herpes simplex virus antibody at control line or control point place.
Preparation method of the present utility model is as follows:
1, anti-cytomegalovirus antibody, Toxoplasma Antibody, rubella virus antibody, anti-herpes simplex virus antibodies or CMV antibody (antigen), toxoplasma antibody (antigen), Detecting Rubella Virus Antibodies In Human Sera (antigen), the commercially available purchase of herpes simplex virus antibody (antigen);
2, the acquisition of standard positive serum:
Collect rubella virus, cytomegalovirus, toxoplasm and herpes simplex virus patient's positive serum, obtain the positive with reference to serum, be decided to be standard positive serum ,-30 ℃ frozen;
The serum of positive patients must be through the Elisa experiment, result positive (colour developing is obvious, positive OD value/negative OD value >=2.1); Through fluorescence immunoassay experiment (IFA), the positive result of result.
3, the point sample of nitrocellulose filter: CMV antibody (antigen) detection line (point), toxoplasma antibody (antigen) detection line (point), Detecting Rubella Virus Antibodies In Human Sera (antigen) detection line (point), herpes simplex virus antibody (antigen) detection line (point) and control line (point) are located on nitrocellulose filter from top to bottom successively, and are shaped with the incubation groove at each detection line (point) and control line (point) surrounding; Be coated with respectively cytomegalovirus antigen (antibody), toxoplasma antigen (antibody), rubella virus antigen (antibody), herpes simplex virus antigens (antibody) on each virus (kind) detection line (point) of nitrocellulose filter; Be coated with respectively from left to right cytomegalovirus antigen, toxoplasma antigen, rubella virus antigen, herpes simplex virus antigens or be coated with respectively from left to right the patient's positive serum containing CMV antibody, toxoplasma antibody, Detecting Rubella Virus Antibodies In Human Sera, herpes simplex virus antibody on each control line (point); Dry;
4, alkali phosphatase enzyme mark or the horseradish peroxidase-labeled of anti-cytomegalovirus antibody, Toxoplasma Antibody, rubella virus antibody, anti-herpes simplex virus antibodies or CMV antibody, toxoplasma antibody, Detecting Rubella Virus Antibodies In Human Sera, herpes simplex virus antibody;
5, prepare the immunodotting kit
With cutting cutter, carrier board is cut into to strip, nitrocellulose filter is sticked on to the carrier board surface; Jointly form four immunodotting kits of family planning with anti-cytomegalovirus antibody, Toxoplasma Antibody, rubella virus antibody, anti-herpes simplex virus antibodies or CMV antibody, toxoplasma antibody, Detecting Rubella Virus Antibodies In Human Sera, herpes simplex virus antibody and alkaline phosphatase substrate or horseradish peroxidase substrate tetramethyl benzidine TMB by enzyme labeling; Described alkaline phosphatase substrate is the bromo-4-of nitroblue tetrazolium and the 5-potpourri that chloro-3-indyl-phosphate BCIP ratio is 1:49.
Four of the disposable fast detecting family plannings of the utility model, simple to operate, cost is low, novel in design, easily by the user, is liked, the universal use of being more convenient for.
The accompanying drawing explanation:
1, Fig. 1 is four associating kit structural representations of point-like enzyme linked immunological spot family planning;
2, Fig. 2 is four associating kit structural representations of wire enzyme linked immunological spot family planning.
Embodiment:
Below by the drawings and specific embodiments, the utility model is further elaborated.
As shown in Figure 1, the utility model is provided with the surface that carrier board 1, nitrocellulose filter 2 stick on carrier board 1, CMV antibody (antigen) check point 3, toxoplasma antibody (antigen) check point 4, Detecting Rubella Virus Antibodies In Human Sera (antigen) check point 5, herpes simplex virus antibody (antigen) check point 6, control point 7 are located at successively on nitrocellulose filter 2 from top to bottom, and are shaped with incubation groove 8 at each detection line point and control point surrounding; Be coated with respectively corresponding antigen (antibody) at CMV antibody (antigen) check point 3, toxoplasma antibody (antigen) check point 4, Detecting Rubella Virus Antibodies In Human Sera (antigen) check point 5, herpes simplex virus antibody (antigen) check point 6 places; Be shaped with from left to right four control point 9-12 at control point 7 places, respectively coated cytomegalovirus antigen, toxoplasma antigen, rubella virus antigen, herpes simplex virus antigens or coated patient's positive serum containing CMV antibody, toxoplasma antibody, Detecting Rubella Virus Antibodies In Human Sera, herpes simplex virus antibody.
As shown in Figure 2, the utility model is provided with the surface that carrier board 1, nitrocellulose filter 2 stick on carrier board 1, CMV antibody (antigen) detection line 3, toxoplasma antibody (antigen) detection line 4, Detecting Rubella Virus Antibodies In Human Sera (antigen) detection line 5, herpes simplex virus antibody (antigen) detection line 6, control line 7 are located at successively on nitrocellulose filter 2 from top to bottom, and are shaped with incubation groove 8 at each detection line and control line surrounding; Be coated with respectively corresponding antigen (antibody) at CMV antibody (antigen) detection line 3, toxoplasma antibody (antigen) detection line 4, Detecting Rubella Virus Antibodies In Human Sera (antigen) detection line 5, herpes simplex virus antibody (antigen) detection line 6 places; Be shaped with from left to right four control line 9-12 at control line 7 places, respectively coated cytomegalovirus antigen, toxoplasma antigen, rubella virus antigen, herpes simplex virus antigens or coated patient's positive serum containing CMV antibody, toxoplasma antibody, Detecting Rubella Virus Antibodies In Human Sera, herpes simplex virus antibody.
The preparation of four associating kits of enzyme linked immunological spot family planning:
1, anti-cytomegalovirus antibody, Toxoplasma Antibody, rubella virus antibody, anti-herpes simplex virus antibodies or CMV antibody (antigen), toxoplasma antibody (antigen), Detecting Rubella Virus Antibodies In Human Sera (antigen), the commercially available purchase of herpes simplex virus antibody (antigen);
2, the preparation of standard positive serum:
Each collects the serum of 50 parts of rubella viruses, cytomegalovirus, toxoplasm and herpes simplex virus positive patients, mixes, and obtains the positive with reference to serum, is decided to be standard positive serum, and-30 ℃ frozen;
The serum of positive patients must be through the Elisa experiment, result positive (colour developing is obvious, positive OD value/negative OD value >=2.1); Through fluorescence immunoassay experiment (IFA), the positive result of result.
3, the point sample of nitrocellulose filter 2: CMV antibody (antigen) detection line (point) 3, toxoplasma antibody (antigen) detection line (point) 4, Detecting Rubella Virus Antibodies In Human Sera (antigen) detection line (point) 5, herpes simplex virus antibody (antigen) detection line (point) 6 and control line (point) 7 are located on nitrocellulose filter 2 from top to bottom successively, and are shaped with the incubation groove at each detection line (point) and control line (point) surrounding; Be coated with respectively corresponding antigen (antibody) on each virus (kind) detection line (point) of nitrocellulose filter 2; Be coated with respectively cytomegalovirus antigen, toxoplasma antigen, rubella virus antigen, herpes simplex virus antigens or coated patient's positive serum containing CMV antibody, toxoplasma antibody, Detecting Rubella Virus Antibodies In Human Sera, herpes simplex virus antibody on four control lines (point) 9-12 be shaped with from left to right at control line (point) 7 places, dry;
4, alkali phosphatase enzyme mark or the horseradish peroxidase-labeled of anti-cytomegalovirus antibody, Toxoplasma Antibody, rubella virus antibody, anti-herpes simplex virus antibodies or CMV antibody, toxoplasma antibody, Detecting Rubella Virus Antibodies In Human Sera, herpes simplex virus antibody;
5, prepare the immunodotting kit
With cutting cutter, carrier board is cut into to strip, nitrocellulose filter 2 is sticked on to carrier board 1 surface; Jointly form four immunodotting kits of family planning with anti-cytomegalovirus antibody, Toxoplasma Antibody, rubella virus antibody, anti-herpes simplex virus antibodies or CMV antibody, toxoplasma antibody, Detecting Rubella Virus Antibodies In Human Sera, herpes simplex virus antibody and alkaline phosphatase substrate (the chloro-potpourri that the 3-indyl-the phosphate ratio is 1:49 of the bromo-4-of nitroblue tetrazolium and 5-) or horseradish peroxidase substrate (tetramethyl benzidine) by enzyme labeling.
The using method of embodiment 2, four associating kits of the utility model enzyme linked immunological spot family planning
1, sample to be measured adopts 0.01mol/L PBS damping fluid to carry out the 1:50 dilution.
2, take out carrier board, draw sample 0.2mL-0.3mL to be measured, join in the incubation groove, in room temperature (18-25) ℃ lower incubation 30min.
3, suck liquid in the incubation groove, with 2mL0.01mol/L PBS damping fluid, rinse each incubation groove 3 times, each 5min.
4, add enzyme conjugates: add respectively 0.2mL-0.3mL enzyme conjugates (adopting two anti-(corresponding antibody) of alkali phosphatase enzyme mark or two anti-(corresponding antibody) of horseradish peroxidase-labeled) in the incubation groove, in 18-25 ℃ of lower incubation 30min.
5, draw liquid in groove, with 2mL0.01mol/L PBS damping fluid, rinse each incubation groove 3 times, each 5min.
6, substrate incubation: add respectively the 0.2mL-0.3mL substrate solution in the incubation groove: potpourri or tetramethyl benzidine that the chloro-3-indyl-phosphate of nitroblue tetrazolium (NBT) and the bromo-4-of 5-(BCIP) ratio is 1:49, in room temperature (18-25) ℃ lower incubation 10min.
7, stop: draw liquid in groove, with distilled water, clean each incubation groove 3 times, each 1min.
Result is judged: by the air-dry rear result of determination of each nitrocellulose filter, black or blue specific band appear in arbitrary detection line, just can be judged to be the positive of corresponding virus.
Claims (1)
1. four associating kits of enzyme linked immunological spot family planning, is characterized in that being provided with carrier board, CMV antibody or antigen detection line or check point, toxoplasma antibody or antigen detection line or check point, Detecting Rubella Virus Antibodies In Human Sera or antigen detection line or check point, herpes simplex virus antibody or antigen detection line or check point, control line or control point, nitrocellulose filter, incubation groove; Nitrocellulose membrane sticks on the surface of carrier board, CMV antibody or antigen detection line or check point, toxoplasma antibody or antigen detection line or check point, Detecting Rubella Virus Antibodies In Human Sera or antigen detection line or check point, herpes simplex virus antibody or antigen detection line or check point, control line or control point are located on nitrocellulose membrane from top to bottom successively, and are shaped with the incubation groove at each detection line or check point and control line or control point surrounding; Coated corresponding antigen or the antibody respectively at CMV antibody or antigen detection line or check point, toxoplasma antibody or antigen detection line or check point, Detecting Rubella Virus Antibodies In Human Sera or antigen detection line or check point, herpes simplex virus antibody or antigen detection line or check point place; Be coated with respectively from left to right cytomegalovirus antigen, toxoplasma antigen, rubella virus antigen, herpes simplex virus antigens or be coated with respectively the patient's positive serum containing CMV antibody, toxoplasma antibody, Detecting Rubella Virus Antibodies In Human Sera, herpes simplex virus antibody at control line or control point place.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201320512939.7U CN203385727U (en) | 2013-08-22 | 2013-08-22 | Enzyme-linked immunospot joint kit for family-planning four items |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201320512939.7U CN203385727U (en) | 2013-08-22 | 2013-08-22 | Enzyme-linked immunospot joint kit for family-planning four items |
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| CN203385727U true CN203385727U (en) | 2014-01-08 |
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| CN201320512939.7U Expired - Fee Related CN203385727U (en) | 2013-08-22 | 2013-08-22 | Enzyme-linked immunospot joint kit for family-planning four items |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103399149A (en) * | 2013-08-22 | 2013-11-20 | 青岛中仁药业有限公司 | Family-planning four-item combined kit employing enzyme linked immunosorbent spot assay |
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2013
- 2013-08-22 CN CN201320512939.7U patent/CN203385727U/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103399149A (en) * | 2013-08-22 | 2013-11-20 | 青岛中仁药业有限公司 | Family-planning four-item combined kit employing enzyme linked immunosorbent spot assay |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140108 Termination date: 20150822 |
|
| EXPY | Termination of patent right or utility model |