CN102507929A - Signal-enhancement type immunochromatographic gold-labeled test strip and preparation method thereof - Google Patents
Signal-enhancement type immunochromatographic gold-labeled test strip and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a signal-enhancement type immunochromatographic gold-labeled test strip and a preparation method thereof. The preparation method of the gold-labeled test strip comprises the following steps of: (1) combining colloidal gold nanometer particles with different particle diameters; (2) preparing a colloidal gold immunity detection probe; (3) preparing a colloidal gold signal enhancement probe; (4) mounting signal-enhancement type test paper; and (5) establishing a detection method of the signal-enhancement type test paper. The signal-enhancement type immunochromatographic gold-labeled test strip, disclosed by the invention, can be applied to field fast ultra-sensitive detection without needing other auxiliary instruments, and has the following advantages that: a detection operation can be finished within 10-15 minutes and a detection result can be obtained; the sensitivity of the traditional test paper detection method can be remarkably improved, and the ultra-sensitive detection of a low-concentration appointed target object which is not realized by the traditional test paper is realized; and as long as an antibody of the immunity identification probe of the test paper is correspondingly replaced, the detection of other target objects can be realized; the specificity is high, the stability is good, the application range is wide, the cost is low and the like; in addition, the signal-enhancement type immunochromatographic gold-labeled test strip and the preparation method thereof, disclosed by the invention, are easy to promote and apply.
Description
Technical field
The invention belongs to the gold test strip bar technical field that food security, inspection and quarantine, environment measuring and disease early diagnosis are used, be specifically related to a kind of signal enhancement mode hypersensitive colloid gold label immuno-chromatographic test paper strip and preparation method thereof.
Background technology
Along with the development of society and the increase of international foreign trade amount, inspection and quarantine and the detection task that food safety detection faced are more and more heavier, and magnanimity test sample amount has proposed new demand to inspection and quarantine and food safety detection.Tradition relies on the detection method of instrument to have the detection sensitivity height, advantages such as testing result good stability.But because large-scale instrument has and costs an arm and a leg, operating personnel must have relevant professional knowledge, and detect shortcoming such as cost costliness and determined to rely on the detection method of large-scale instrument in various basic units and on-the-spot the detection, extensively to promote the use of.Detection method based on immuno analytical method has been alleviated the severe situation that current detection faced to a certain extent.Like enzyme linked immunosorbent assay analysis method, have the detection sensitivity height, detect characteristics such as cost is cheap relatively.The appearance of colloidal gold colloidal gold detection test paper strip realizes the fast detecting of object for great innovation was arranged on detection time in less than 1 hour time range.Simultaneously, because gold test strip bar detection technique does not need other complicated instruments in testing process, only need to judge the testing result of test strips through visual inspection.Under such condition, the detection sensitivity of test strips is significant for the judgement of testing result.But the development of gold test strip bar so far, and the optimization of its detection sensitivity is one of research emphasis of test strips related detecting method always.
Summary of the invention
The present invention is directed to the present Research that the sensitivity that has gold test strip bar and related detecting method thereof now remains further raising; Utilize the specific surface effect of nano material; Carry out the research that gold test strip bar detection signal strengthens; A kind of signal enhancement mode immunochromatography gold test strip bar and preparation method thereof is provided, has realized the target detection thing is carried out the hypersensitive field quick detection.
The concrete technical scheme of the present invention is following:
A kind of signal enhancement mode hypersensitive immunochromatography gold test strip bar comprises plastics lining board; Be pasted with the mother glass tunica fibrosa on the said plastics lining board respectively successively; Immunological probe gold mark pad, amplification of signal gold mark pad, five kinds of adhesive matter of nitrocellulose membrane and water adsorption glass tunica fibrosa; The length of lap is 2 mm between the adjacent adhesive matter, dry encapsulation, and 4 ℃～normal temperature condition of temperature is preserved down.
A kind of preparation method's of signal enhancement mode immunochromatography gold test strip bar concrete preparation manipulation step is following:
(1) preparation of target detection thing gold-marking immunity probe
Choose the colloidal gold solution that particle diameter is 30～40nm, using concentration is the sal tartari (K of 0.1 M
2CO
3) solution regulates pH value to 8.5～9.5 of colloidal gold solution, must regulate colloidal gold solution A afterwards;
The anti-target detection thing of rabbit antibody-solutions and the concentration that with concentration is 1mg～2 mg/mL is that bovine serum albumin(BSA) (BSA) solution of 1 mg～2 mg/mL mixes with mol ratio 1:100, mixed solution;
Get mixed solution 10 μ L and join among the 1 mL adjusting back colloidal gold solution A oscillating reactions 15min under the room temperature;
Under 4 ℃ of conditions of temperature,, remove unconjugated antibody of supernatant and bovine serum albumin(BSA) (BSA) with centrifugal 20 min of the speed of 10000 rpm;
The centrifugal deposition that obtains is scattered in the phosphate buffer solution (PBS) again, obtains target detection thing gold-marking immunity probe; (2) preparation of gold mark amplification of signal probe
Choose the colloidal gold solution that particle diameter is 15～20 nm, using concentration is the sal tartari (K of 0.1 M
2CO
3) solution regulates pH value to 8.5～9.5 of colloidal gold solution, must regulate colloidal gold solution B afterwards;
Is that bovine serum albumin(BSA) (BSA) antibody-solutions of 1mg/mL joins and regulates among the colloidal gold solution B of back with the mark ratio of 1:1000～1:3000 with concentration, reacts 15min under the room temperature; Under 4 ℃ of conditions of temperature, behind centrifugal 20 min of the speed of 10000 rpm, remove unconjugated bovine serum albumin(BSA) (BSA) antibody in the supernatant;
The centrifugal deposition that obtains is scattered in the phosphate buffer solution (PBS) again, obtains gold mark amplification of signal probe;
(3) preparation of target detection thing immunological probe gold mark pad and target detection thing amplification of signal gold mark pad
Target detection thing gold-marking immunity probe and gold mark amplification of signal probe are sprayed to respectively on two glass fibre membranes, obtain target detection thing immunological probe gold mark pad and target detection thing amplification of signal gold mark pad;
(4) has the preparation of the nitrocellulose membrane of detection line and line of reference
The line that on a nitrocellulose membrane, uses concentration to draw two line parallels respectively as the goat anti-rabbit antibody solution of 1mg/mL as target detection thing-ovalbumin conjugate solution and the concentration of 1mg/mL; Target detection thing-ovalbumin conjugate is rule as detection line, and goat anti-rabbit antibody solution is scribed ss line of reference;
(5) preparation of signal enhancement mode immunochromatography gold test strip bar
Nitrocellulose membrane and five kinds of adhesive matter of water adsorption glass tunica fibrosa of mother glass tunica fibrosa, the golden mark pad of target detection thing immunological probe, target detection thing amplification of signal gold mark being filled up, have detection line and line of reference paste on the plastics lining board respectively successively; The length of the lap between the adjacent adhesive matter is 2 mm, obtains signal enhancement mode super sensitivity detection gold test strip bar, dry encapsulation, and 4 ℃～normal temperature condition of temperature is preserved down.
The anti-target detection thing of rabbit in the said step 1 antibody is rabbit anti-bisphenol A antibody, anti-phthalic ester antibody of rabbit or the anti-Lay of rabbit cordoba amine antibody.
Useful technique effect of the present invention embodies in the following areas:
1, the present invention is applied to big this distinguishing feature of nano particle specific surface area the foundation and the corresponding detecting method of traditional gold test strip bar; Set up the signal enhancement mode colloidal gold mark test paper and relevant detection method that can be used for on-the-spot fast super sensitivity detection, realized that the fast qualitative of target detection thing detects and the target detection thing is carried out detection by quantitative through the reduction of colour developing degree on the test strip through the naked eyes Direct observation.Need not other supplementary instrument equipment, testing result can accomplished and obtain to testing in 10-15 minute;
2, the present invention the most the part of core be the design through the signal enhancement function, can significantly improve the sensitivity of traditional test strips detection method, realize the super sensitivity detection of the low concentration specific objective thing that traditional test strips can't realize;
3, the present invention is universal immunochromatography colloidal gold mark test paper, only needs the antibody that immune recognizing probe in the test strips is used to carry out corresponding replacement, can realize the detection of other objects;
4, the present invention also has the specificity height, good stability, and applied range, with low cost, the method for manipulating simply is easy to apply.Port such as customs, airport that can be applicable to hospital, family and food security and inspection and quarantine etc. needs field quick detection mechanism.
Description of drawings
Fig. 1 is the side structure synoptic diagram of signal enhancement metal mark test strips.
Sequence number among Fig. 1: mother glass tunica fibrosa 1, immunological probe gold mark pad 2, amplification of signal gold are marked pad 3, are had nitrocellulose membrane 4, water adsorption glass tunica fibrosa 5, plastics lining board 10, detection line 11, the line of reference 12 of detection line and line of reference.
Fig. 2 is the positive testing result synoptic diagram of signal enhancement metal mark test strips.
Embodiment
Below in conjunction with embodiment, the present invention is done to describe further.
A kind of preparation method's concrete operations step of signal enhancement mode bisphenol-A immunochromatography gold test strip bar is following:
(1) preparation of rabbit anti-bisphenol A gold-marking immunity probe
Choose the colloidal gold solution that particle diameter is 40nm, using concentration is the sal tartari (K of 0.1 M
2CO
3) solution regulates the pH value to 8.5 of colloidal gold solution, must regulate colloidal gold solution A afterwards;
Rabbit anti-bisphenol A antibody-solutions and the concentration that with concentration is 1mg/mL is that bovine serum albumin(BSA) (BSA) solution of 1 mg/mL mixes with mol ratio 1:100, mixed solution;
Get mixed solution 10 μ L and join among the 1 mL adjusting back colloidal gold solution A oscillating reactions 15min under the room temperature;
Under 4 ℃ of conditions of temperature,, remove unconjugated antibody of supernatant and bovine serum albumin(BSA) (BSA) with centrifugal 20 min of the speed of 10000 rpm;
The centrifugal deposition that obtains is scattered in the phosphate buffer solution (PBS) again, obtains rabbit anti-bisphenol A gold-marking immunity probe; (2) preparation of rabbit anti-bisphenol A gold mark amplification of signal probe
Choose the colloidal gold solution that particle diameter is 20 nm, using concentration is the sal tartari (K of 0.1 M
2CO
3) solution regulates the pH value to 8.5 of colloidal gold solution, must regulate colloidal gold solution B afterwards;
Is that bovine serum albumin(BSA) (BSA) antibody-solutions of 1mg/mL joins and regulates among the colloidal gold solution B of back with the mark ratio of 1:1000 with concentration, reacts 15min under the room temperature; Under 4 ℃ of conditions of temperature, behind centrifugal 20 min of the speed of 10000 rpm, remove unconjugated bovine serum albumin(BSA) (BSA) antibody in the supernatant;
The centrifugal deposition that obtains is scattered in the phosphate buffer solution (PBS) again, obtains rabbit anti-bisphenol A gold mark amplification of signal probe;
(3) preparation of rabbit anti-bisphenol A immunological probe gold mark pad and rabbit anti-bisphenol A amplification of signal gold mark pad
Rabbit anti-bisphenol A gold-marking immunity probe and rabbit anti-bisphenol A gold mark amplification of signal probe are sprayed to respectively on two glass fibre membranes, obtain rabbit anti-bisphenol A immunological probe gold mark pad and rabbit anti-bisphenol A amplification of signal gold mark pad;
(4) has the preparation of the nitrocellulose membrane of detection line and line of reference
The line that on a nitrocellulose membrane, uses concentration to draw two line parallels respectively as the goat anti-rabbit antibody solution of 1mg/mL as bisphenol-A-ovalbumin conjugate solution and the concentration of 1mg/mL; Rabbit anti-bisphenol A-ovalbumin conjugate is rule as detection line, and goat anti-rabbit antibody solution is scribed ss line of reference;
(5) preparation of signal enhancement mode bisphenol-A immunochromatography gold test strip bar
Referring to Fig. 1, on plastics lining board 10, paste mother glass tunica fibrosa 1, rabbit anti-bisphenol A immunological probe gold mark pad 2, golden nitrocellulose membrane 4 and the water adsorption glass tunica fibrosa 5 of marking pad 3, having detection line and line of reference of rabbit anti-bisphenol A amplification of signal respectively successively; The length of the lap between the adjacent adhesive matter is 2 mm, obtains signal enhancement mode bisphenol-A super sensitivity detection gold test strip bar, dry encapsulation, and 4 ℃～normal temperature condition of temperature is preserved down.
During use, the mother glass tunica fibrosa end of signal enhancement mode rabbit anti-bisphenol A gold test strip bar is inserted in the solution to be detected, takes out the testing result that detects by an unaided eye after waiting for 10 min.If developing the color with the contrast band simultaneously, the test strip on the film shows that testing result is effective; Compare with the test strip of blank group, if the test strip color of test set shoals and shows and contain object rabbit anti-bisphenol A in the solution to be detected, rabbit anti-bisphenol A concentration is high more in the shallow more detection solution that develops the color, positive result; As have only test strip colour developing and the contrast band does not develop the color, show that testing result is invalid.
Embodiment 2:
A kind of preparation method's concrete operations step of signal enhancement mode phthalic ester immunochromatography gold test strip bar is following:
(1) preparation of the anti-phthalic ester gold-marking immunity of rabbit probe
Choose the colloidal gold solution that particle diameter is 30nm, using concentration is the pH value to 9.5 of sal tartari (K2CO3) the solution adjusting colloidal gold solution of 0.1 M, must regulate back colloidal gold solution A;
The anti-phthalic ester antibody-solutions of rabbit and the concentration that with concentration is 2mg/mL is that bovine serum albumin(BSA) (BSA) solution of 2mg/mL mixes with mol ratio 1:100, mixed solution;
Get mixed solution 10 μ L and join among the 1 mL adjusting back colloidal gold solution A oscillating reactions 15min under the room temperature;
Under 4 ℃ of conditions of temperature,, remove unconjugated antibody of supernatant and bovine serum albumin(BSA) (BSA) with centrifugal 20 min of the speed of 10000 rpm;
The centrifugal deposition that obtains is scattered in the phosphate buffer solution (PBS) again, obtains the anti-phthalic ester gold-marking immunity of rabbit probe;
(2) preparation of the anti-phthalic ester gold of rabbit mark amplification of signal probe
Choose the colloidal gold solution that particle diameter is 15nm, using concentration is the pH value to 9.5 of sal tartari (K2CO3) the solution adjusting colloidal gold solution of 0.1 M, must regulate back colloidal gold solution B;
Is that bovine serum albumin(BSA) (BSA) antibody-solutions of 1mg/mL joins and regulates among the colloidal gold solution B of back with the mark ratio of 1:3000 with concentration, reacts 15min under the room temperature; Under 4 ℃ of conditions of temperature, behind centrifugal 20 min of the speed of 10000 rpm, remove unconjugated bovine serum albumin(BSA) (BSA) antibody in the supernatant;
The centrifugal deposition that obtains is scattered in the phosphate buffer solution (PBS) again, obtains the anti-phthalic ester gold of rabbit mark amplification of signal probe;
(3) preparation of the anti-phthalic ester immunological probe gold of rabbit mark pad and the anti-phthalic ester amplification of signal gold of rabbit mark pad
Anti-phthalic ester immunological probe of rabbit and the anti-phthalic ester amplification of signal of rabbit probe are sprayed to respectively on two glass fibre membranes, obtain the anti-phthalic ester immunological probe gold of rabbit mark pad and the anti-phthalic ester amplification of signal gold of rabbit mark pad respectively;
(4) has the preparation of the nitrocellulose membrane of detection line and line of reference
On a nitrocellulose membrane, using concentration is the line that the anti-phthalic ester of the rabbit-ovalbumin conjugate solution of 0.5 mg/mL and goat anti-rabbit antibody solution that concentration is 0.5 mg/mL are drawn two line parallels respectively; The anti-phthalic ester of rabbit-ovalbumin conjugate is rule as detection line, and goat anti-rabbit antibody solution is scribed ss line of reference;
(5) preparation of the anti-phthalic ester immunochromatography of signal enhancement mode rabbit gold test strip bar
Referring to Fig. 1, on plastics lining board 1, paste mother glass tunica fibrosa 2, the anti-phthalic ester immunological probe gold of rabbit mark pad 3, golden nitrocellulose membrane 5 and the water adsorption glass tunica fibrosa 6 of marking pad 4, having detection line and line of reference of the anti-phthalic ester amplification of signal of rabbit respectively successively; The length of the lap between the adjacent adhesive matter is 2 mm, obtains signal enhancement mode phthalic ester super sensitivity detection gold test strip bar, dry encapsulation, and 4 ℃～normal temperature condition of temperature is preserved down.
Preparation method's concrete operations step of the anti-Lay of a kind of signal enhancement mode rabbit cordoba amine immunochromatography gold test strip bar is following:
(1) preparation of the anti-Lay of rabbit cordoba amine gold-marking immunity probe
Choose the colloidal gold solution that particle diameter is 30nm, using concentration is the sal tartari (K of 0.1 M
2CO
3) solution regulates the pH value to 9.5 of colloidal gold solution, must regulate colloidal gold solution A afterwards;
The anti-Lay of rabbit cordoba amine antibody-solutions and the concentration that with concentration is 2mg/mL is that bovine serum albumin(BSA) (BSA) solution of 2mg/mL mixes with mol ratio 1:100, mixed solution;
Get mixed solution 10 μ L and join among the 1 mL adjusting back colloidal gold solution A oscillating reactions 15min under the room temperature;
Under 4 ℃ of conditions of temperature,, remove unconjugated antibody of supernatant and bovine serum albumin(BSA) (BSA) with centrifugal 20 min of the speed of 10000 rpm;
The centrifugal deposition that obtains is scattered in the phosphate buffer solution (PBS) again, obtains the anti-Lay of rabbit cordoba amine gold-marking immunity probe;
(2) preparation of the anti-Lay of rabbit cordoba amine gold mark amplification of signal probe
Choose the colloidal gold solution that particle diameter is 15nm, using concentration is the sal tartari (K of 0.1 M
2CO
3) solution regulates the pH value to 9.5 of colloidal gold solution, must regulate colloidal gold solution B afterwards;
Is that bovine serum albumin(BSA) (BSA) antibody-solutions of 1mg/mL joins and regulates among the colloidal gold solution B of back with the mark ratio of 1:3000 with concentration, reacts 15min under the room temperature; Under 4 ℃ of conditions of temperature, behind centrifugal 20 min of the speed of 10000 rpm, remove unconjugated bovine serum albumin(BSA) (BSA) antibody in the supernatant;
The centrifugal deposition that obtains is scattered in the phosphate buffer solution (PBS) again, obtains the anti-Lay of rabbit cordoba amine gold mark amplification of signal probe;
(3) preparation of the anti-Lay of rabbit cordoba amine immunological probe gold mark pad and the anti-Lay of rabbit cordoba amine amplification of signal gold mark pad
The anti-Lay of rabbit cordoba amine immunological probe and the anti-Lay of rabbit cordoba amine amplification of signal probe are sprayed to respectively on two glass fibre membranes, obtain the anti-Lay of rabbit cordoba amine immunological probe gold mark pad and the anti-Lay of rabbit cordoba amine amplification of signal gold mark pad respectively;
(4) has the preparation of the nitrocellulose membrane of detection line and line of reference
On a nitrocellulose membrane, using concentration is the line that the anti-Lay of the rabbit cordoba amine-ovalbumin conjugate solution of 0.5 mg/mL and goat anti-rabbit antibody solution that concentration is 0.5 mg/mL are drawn two line parallels respectively; The anti-Lay of rabbit cordoba amine-ovalbumin conjugate is rule as detection line, and goat anti-rabbit antibody solution is scribed ss line of reference;
(5) preparation of the anti-Lay of signal enhancement mode rabbit cordoba amine immunochromatography gold test strip bar
Referring to Fig. 1, on plastics lining board 1, paste mother glass tunica fibrosa 2, the anti-Lay of rabbit cordoba amine immunological probe gold mark pad 3, golden nitrocellulose membrane 5 and the water adsorption glass tunica fibrosa 6 of marking pad 4, having detection line and line of reference of the anti-Lay of rabbit cordoba amine amplification of signal respectively successively; The length of the lap between the adjacent adhesive matter is 2 mm, obtains the anti-Lay of signal enhancement mode rabbit cordoba amine super sensitivity detection gold test strip bar, dry encapsulation, and 4 ℃～normal temperature condition of temperature is preserved down.
Embodiment 4:
Referring to Fig. 2, wherein sequence number 2 is the mother glass tunica fibrosa, and length is 18mm; Sequence number 3 is a gold-marking immunity recognizing probe pad, and length is 4 mm; Sequence number 4 is a gold mark amplification of signal probe pads, and length is 4 mm; Sequence number 5 is a nitrocellulose membrane, and length is 20 mm; Sequence number 6 is an adsorptive pads, and length is 18 mm, and sequence number 11 is a detection line, and width is 1mm, and sequence number 12 is a line of reference, and width is 1 mm.Arrow express liquid capillary action direction among the figure.
Fig. 2 carries out in the object testing process basis for estimation of testing result for utilizing signal enhanced colloidal gold mark test paper.How through the testing result of signal enhancement metal mark test strips the content of object in the test sample to be judged, be the important component part of detection method.Concrete testing result basis for estimation is as follows:
The first from left is the negative result synoptic diagram of signal enhancement metal mark test strips among Fig. 2,
The second from left is the weak positive test symbol synoptic diagram of signal enhancement metal mark test strips among Fig. 2,
Fig. 2 right-of-center in political views one is the positive test symbol synoptic diagram of signal enhancement metal mark test strips,
Fig. 2 right-of-center in political views two is invalid signal enhancement metal mark test strips testing result synoptic diagram.
Claims (3)
1. signal enhancement mode immunochromatography gold test strip bar; It is characterized in that: the gold test strip bar comprises plastics lining board; Be pasted with the mother glass tunica fibrosa on the said plastics lining board respectively successively; Immunological probe gold mark pad, amplification of signal gold mark pad, five kinds of adhesive matter of nitrocellulose membrane and water adsorption glass tunica fibrosa; The length of lap is 2 mm between the adjacent adhesive matter, dry encapsulation, and 4 ℃～normal temperature condition of temperature is preserved down.
2. the preparation method of a signal enhancement mode immunochromatography gold test strip bar is characterized in that concrete preparation manipulation step is following:
(1) preparation of target detection thing gold-marking immunity probe
Choose the colloidal gold solution that particle diameter is 30～40nm, using concentration is the sal tartari (K of 0.1 M
2CO
3) solution regulates pH value to 8.5～9.5 of colloidal gold solution, must regulate colloidal gold solution A afterwards;
The anti-target detection thing of rabbit antibody-solutions and the concentration that with concentration is 1mg～2 mg/mL is that bovine serum albumin(BSA) (BSA) solution of 1 mg～2 mg/mL mixes with mol ratio 1:100, mixed solution;
Get mixed solution 10 μ L and join among the 1 mL adjusting back colloidal gold solution A oscillating reactions 15min under the room temperature;
Under 4 ℃ of conditions of temperature,, remove unconjugated antibody of supernatant and bovine serum albumin(BSA) (BSA) with centrifugal 20 min of the speed of 10000 rpm;
The centrifugal deposition that obtains is scattered in the phosphate buffer solution (PBS) again, obtains target detection thing gold-marking immunity probe; (2) preparation of gold mark amplification of signal probe
Choose the colloidal gold solution that particle diameter is 15～20 nm, using concentration is the sal tartari (K of 0.1 M
2CO
3) solution regulates pH value to 8.5～9.5 of colloidal gold solution, must regulate colloidal gold solution B afterwards;
Is that bovine serum albumin(BSA) (BSA) antibody-solutions of 1mg/mL joins and regulates among the colloidal gold solution B of back with the mark ratio of 1:1000～1:3000 with concentration, reacts 15min under the room temperature; Under 4 ℃ of conditions of temperature, behind centrifugal 20 min of the speed of 10000 rpm, remove unconjugated bovine serum albumin(BSA) (BSA) antibody in the supernatant;
The centrifugal deposition that obtains is scattered in the phosphate buffer solution (PBS) again, obtains gold mark amplification of signal probe;
(3) preparation of target detection thing immunological probe gold mark pad and target detection thing amplification of signal gold mark pad
Target detection thing gold-marking immunity probe and gold mark amplification of signal probe are sprayed to respectively on two glass fibre membranes, obtain target detection thing immunological probe gold mark pad and target detection thing amplification of signal gold mark pad;
(4) has the preparation of the nitrocellulose membrane of detection line and line of reference
The line that on a nitrocellulose membrane, uses concentration to draw two line parallels respectively as the goat anti-rabbit antibody solution of 1mg/mL as target detection thing-ovalbumin conjugate solution and the concentration of 1mg/mL; Target detection thing-ovalbumin conjugate is rule as detection line, and goat anti-rabbit antibody solution is scribed ss line of reference;
(5) preparation of signal enhancement mode immunochromatography gold test strip bar
Nitrocellulose membrane and five kinds of adhesive matter of water adsorption glass tunica fibrosa of mother glass tunica fibrosa, the golden mark pad of target detection thing immunological probe, target detection thing amplification of signal gold mark being filled up, have detection line and line of reference paste on the plastics lining board respectively successively; The length of the lap between the adjacent adhesive matter is 2 mm, obtains signal enhancement mode super sensitivity detection gold test strip bar, dry encapsulation, and 4 ℃～normal temperature condition of temperature is preserved down.
3. the preparation method of a kind of signal enhancement mode immunochromatography gold test strip bar according to claim 2; It is characterized in that: the anti-target detection thing of the rabbit in the said step 1 antibody is rabbit anti-bisphenol A antibody, anti-phthalic ester antibody of rabbit or the anti-Lay of rabbit cordoba amine antibody.
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