CN101221179A - Bladder cancer detecting test paper - Google Patents
Bladder cancer detecting test paper Download PDFInfo
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- CN101221179A CN101221179A CNA2007101731339A CN200710173133A CN101221179A CN 101221179 A CN101221179 A CN 101221179A CN A2007101731339 A CNA2007101731339 A CN A2007101731339A CN 200710173133 A CN200710173133 A CN 200710173133A CN 101221179 A CN101221179 A CN 101221179A
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- test paper
- bladder cancer
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- antibody
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Abstract
The invention relates to a bladder cancer detection test paper, which consists of a sample pad, a colloidal gold pad, a nitrocellulose membrane, an absorbent paper and a bottom plate, wherein a detection area of urinary fibronectin-bovine serum albumin conjugate and a control area of anti-mouse secondary antibody are arranged on the nitrocellulose membrane, and a colloidal-gold anti-uric fibronectin antibody is arranged on the colloidal gold pad, wherein, the colloidal-gold anti-uric fibronectin antibody is a colloid with the particle size of 5 to 40nm. The invention is characterized by rapidity, high sensitivity, strong specificity, good stability, simple and convenient operation and no need of any instrument or equipment, furthermore, the judgment of the result is intuitive, reliable and easy to master, and the invention is applicable to the diagnosis of bladder cancer diseases and the self-monitor of the disease conditions.
Description
Technical field
The invention belongs to biological technical field, particularly relate to a kind of detection test paper of carcinoma of urinary bladder.
Background technology
(Bladder transitional cell carcinoma BTCC) is one of common cancer of China to TCCB, is the 5th of the male sex, the 6th～7 of women.Find early, diagnosing bladder cancer to the treatment and prognosis significant effects is arranged.Carcinoma of urinary bladder diagnosis at present mainly depends on cystoscope, but cystoscopy is the operation of a kind of invasive, the diagnosis and treatment process complexity, wastes time and energy, and expense is higher, and patient's misery is difficult to tolerance, and a lot of patients have lost the chance of timely diagnosis, treatment because fear cystoscopy.For this reason, find a kind of easy, do not have a wound, cheap and diagnostic method is extremely urgent efficiently.
(Fibronectin FN) is a kind of widely new carcinoma of urinary bladder tumor marker of research to the urine fibronectin.Studies show that the diagnosis index of urine FN as wellability BTCC, the sensitivity of diagnosis is 76.47%～100%, and specificity is 87.22%, and accuracy of diagnosis is 86.96%.At present, the clinical detection of urine FN mainly adopts the ELISA method to detect, and remain in following problem: 1, the ELISA kit of Cai Yonging is different manufacturers production, lacks standardization.2, step is more loaded down with trivial details, and the result is subject to the influence of operator's technical merit etc., and uncertain factor is more.3, the ELISA method needs the operator to possess certain molecular biology technical ability, is difficult for carrying out at basic hospital.4, the EIISA method detects and not only requires the operator to have the also essential microplate reader of certain operative skill, so patient can't monitor the state of an illness of oneself at home.
(Gold immunochromatography assay GICA) is a kind of immobilon-p immune analysis method that colloid gold immune technology and chromatography technology are combined that rises the nineties in 20th century to colloidal nano gold immune chromatography method.Its principle is a certain district band that special antigen or antibody is fixed in nitrocellulose membrane earlier, after this dry nitrocellulose filter one end immerses sample (as urine, blood, saliva etc.), because capillarity, sample just can move forward along nitrocellulose membrane with golden labeling antibody or the antigen on the glue gold pad, when moving to when being fixed with antigen or antibody regional, antigen and antibody specificity association reaction take place in the system, this reaction is assembled collaurum somewhere around here and is demonstrated aubergine, thereby realizes specific immunodiagnosis.The colloidal gold test that is gone out by this technological development has been widely used in clinical diagnosis and drug test, and its kind comprises: the early pregnancy class detects test paper (detecting HCG, LH, FSH etc.), infectious disease detects test paper (detecting HIV, HCV, HBV, HBsAg etc.), drugs, medicament residue detection test paper (detecting morphine, cocaine, opium, hemp, head-shaking pill, clenbuterol hydrochloride etc.) etc.
Summary of the invention
Technical matters to be solved
Technical matters to be solved by this invention provides a kind of detection test paper of carcinoma of urinary bladder, with overcome existing carcinoma of urinary bladder detect in the diagnosis and treatment process complexity, waste time and energy, expense is higher, and the state of an illness is difficult to defectives such as self-monitoring.
Technical scheme
One of technical scheme of the present invention provides a kind of bladder cancer detecting test paper, form by sample pad, glue gold pad, nitrocellulose membrane, thieving paper and base plate, the detection zone of urine fibronectin-bovine serum albumin conjugate and the check plot that anti-mouse two resists are arranged on described nitrocellulose membrane, the anti-urine of gold mark fibronectin antibody is arranged on described glue gold pad, wherein, the anti-urine of described gold mark fibronectin antibody is the colloid of particle diameter 5-40nm.
One of preferred version of above-mentioned bladder cancer detecting test paper is that the anti-urine of described gold mark fibronectin antibody prepares with following method: with the HAuCl of 100mL 0.01%
4Solution is heated to and boils, the sodium citrate solution that under vigorous stirring, adds 0.5-5mL rapidly, close thermal source behind the maintenance fluidized state reaction 5-30min and promptly get colloidal gold solution, the anti-urine of mixing fibronectin antibody is with colloidal gold solution and it is combined, and obtains stable golden labeling antibody.
Two of the preferred version of above-mentioned bladder cancer detecting test paper is that described glue gold pad is made up of glass fibre and the golden labeling antibody of fixing on it.
Three of the preferred version of above-mentioned bladder cancer detecting test paper is that described sample pad is the water absorptivity glass layer.
Four of the preferred version of above-mentioned bladder cancer detecting test paper is, has at least one nature controlling line on the described test paper, and its colour developing expression test paper is effective.
Two of technical scheme of the present invention provides a kind of preparation method of bladder cancer detecting test paper, in turn includes the following steps:
1) with the HAuCl of 100mL 0.01%
4Solution is heated to and boils, and adds the sodium citrate solution of 0.5-5mL under vigorous stirring rapidly, closes thermal source behind the maintenance fluidized state reaction 5-30min and promptly gets colloidal gold solution;
2) mix anti-urine fibronectin antibody with colloidal gold solution and it is combined, obtain stable golden labeling antibody;
3) golden labeling antibody is fixed on the glass fibre, makes glue gold pad;
4) with sample pad, polyester film, glue gold pad, cellulose nitrate layer and thieving paper, overlapping successively linking to each other sticks on the base plate, is assembled into the detection test paper.
Three of technical scheme of the present invention provides a kind of above-mentioned application of bladder cancer detecting test paper in detection, being about to fluid sample drips in described test paper sample pad one side, by antigen and antibody specificity association reaction, make colloidal gold aggregation and develop the color, thereby realize that the result judges.
One of preferred version of the application of above-mentioned bladder cancer detecting test paper in detection is that the detectability of described test paper is lower than 200ug/L urine fibronectin.
Two of the preferred version of the application of above-mentioned bladder cancer detecting test paper in detection is, has at least one nature controlling line on the described test paper, and its colour developing expression test paper is effective.
Three of the preferred version of the application of above-mentioned bladder cancer detecting test paper in detection is that described fluid sample is a urine.
The present invention can realize in the following way, also can use the similar fashion of effect same:
Described test paper assembles (as shown in Figure 1) by overlapping successively continuous the sticking on the base plate of order from left to right by inhaling glass layer (glue gold pad), cellulose nitrate rete (NC film) and the absorbent paper layer of urinating glass layer (sample pad), polyester rete, being fixed with golden labeling antibody.The gold labeling antibody is the compound of colloid gold particle and anti-FN antibodies; Be marked with T line (p-wire) and C line (nature controlling line) on the nitrocellulose membrane, the T line is the FN-BSA conjugate, and the C line is two anti-for the anti-mouse of bag quilt.After treating patient's urine sample dropping, if the colour developing of T line, the C line also develops the color, and the result who obtains is negative, shows that patient does not have the possibility of suffering from carcinoma of urinary bladder.If the T line does not develop the color, the colour developing of C line, the result who obtains is positive, shows that patient may suffer from carcinoma of urinary bladder.If nature controlling line (C line) does not develop the color, the expression test paper lost efficacy (as shown in Figure 2).
Beneficial effect
The present invention adopts colloidal gold immunochromatographimethod technical design, a kind of test paper that is used for detecting patient's urine sample FN content of exploitation, for the clinical diagnosis of carcinoma of urinary bladder provides reliable foundation.That the colloidal gold immunochromatographimethod detection method not only has is quick, highly sensitive, high specificity, good stability, easy and simple to handle, need not any instrument and equipment, and the result judge intuitive and reliable, grasped by the personnel of grass-roots unit easily, also make things convenient for simultaneously the self-monitoring of patient to the state of an illness.Whether the present invention adopts the colloidal gold immunochromatographimethod technology in the detection of carcinoma of urinary bladder, only need patient's several urines, just can check out the content of patient's FN to exceed standard, and for diagnosis of bladder cancer provides reliable foundation, is expected to become the new method of diagnosing bladder cancer.
Description of drawings
Fig. 1 is the bladder cancer detecting test paper structural representation
Fig. 2 detects synoptic diagram for bladder cancer detecting test paper
Fig. 3 is the TEM figure of collaurum
Embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, as molecular labeling operation manual or clinical detection operation manual, or the condition of advising according to manufacturer.
Embodiment 1
1, adopt classical Frens legal system to be equipped with the colloidal gold solution that particle diameter is 5-40nm: with the HAuCl of 100mL 0.01%
4Solution is heated to and boils, and adds the sodium citrate solution of 0.5-5mL under vigorous stirring rapidly, keeps closing thermal source behind the fluidized state reaction 5-30min, promptly gets the shiny red transparent colloid.
2, select suitable pH value (6-10), antibody is combined securely with collaurum, prepare stable golden labeling antibody.
3, with golden labeling antibody specking on glass fibre, preparation plastic emitting gold pad.Two anti-side specking FN-BSA conjugate of nitrocellulose membrane (NC film) and anti-mouse then is respectively as T line and C line.
4, sample pad (length is 35mm), glue gold pad (length is 10mm), NC film (length is 25mm) and thieving paper (length is 20mm) are contained on the base plate according to from left to right der group, and cut into the wide test strips of 4mm that is with cutting knife.
Test paper detecting method:
1, the sample to be tested of getting 50ul is added in the sample application zone of test strips;
2, in 10min postscript observed and recorded testing result.The result who observes behind the 20min is invalid.
The judgement of testing result:
1, positive findings: an aubergine band only on the nature controlling line of test strips, occurs.
2, negative findings: on the p-wire of test strips and nature controlling line position, an aubergine band respectively occurs.
3, null result: the aubergine band on the nature controlling line of test strips, do not occur.
Claims (10)
1. bladder cancer detecting test paper, form by sample pad, glue gold pad, nitrocellulose membrane, thieving paper and base plate, the detection zone of urine fibronectin-bovine serum albumin conjugate and the check plot that anti-mouse two resists are arranged on described nitrocellulose membrane, the anti-urine of gold mark fibronectin antibody is arranged on described glue gold pad, wherein, the anti-urine of described gold mark fibronectin antibody is the colloid of particle diameter 5-40nm.
2. bladder cancer detecting test paper according to claim 1 is characterized in that, the anti-urine of described gold mark fibronectin antibody prepares with following method: with the HAuCl of 100mL 0.01%
4Solution is heated to and boils, the sodium citrate solution that under vigorous stirring, adds 0.5-5mL rapidly, close thermal source behind the maintenance fluidized state reaction 5-30min and promptly get colloidal gold solution, the anti-urine of mixing fibronectin antibody is with colloidal gold solution and it is combined, and obtains stable golden labeling antibody.
3. bladder cancer detecting test paper according to claim 1 is characterized in that, described glue gold pad is made up of glass fibre and the golden labeling antibody of fixing on it.
4. bladder cancer detecting test paper according to claim 1 is characterized in that, described sample pad is the water absorptivity glass layer.
5. bladder cancer detecting test paper according to claim 1 is characterized in that, has at least one nature controlling line on the described test paper, and its colour developing expression test paper is effective.
6. the preparation method of a bladder cancer detecting test paper in turn includes the following steps:
1) with the HAuCl of 100mL 0.01%
4Solution is heated to and boils, and adds the sodium citrate solution of 0.5-5mL under vigorous stirring rapidly, closes thermal source behind the maintenance fluidized state reaction 5-30min and promptly gets colloidal gold solution;
2) mix anti-urine fibronectin antibody with colloidal gold solution and it is combined, obtain stable golden labeling antibody;
3) golden labeling antibody is fixed on the glass fibre, makes glue gold pad;
4) with sample pad, polyester film, glue gold pad, cellulose nitrate layer and thieving paper, overlapping successively linking to each other sticks on the base plate, is assembled into the detection test paper.
7. the application of the described bladder cancer detecting test paper of claim 1 in detection is about to fluid sample and drips in described test paper sample pad one side, by antigen and antibody specificity association reaction, makes colloidal gold aggregation and develops the color, thereby realize result's judgement.
8. the application of bladder cancer detecting test paper according to claim 7 in detection is characterized in that, the detectability of described test paper is lower than 200ug/L urine fibronectin.
9. the application of bladder cancer detecting test paper according to claim 7 in detection is characterized in that, has at least one nature controlling line on the described test paper, and its colour developing expression test paper is effective.
10. the application of bladder cancer detecting test paper according to claim 7 in detection is characterized in that described fluid sample is a urine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007101731339A CN101221179A (en) | 2007-12-26 | 2007-12-26 | Bladder cancer detecting test paper |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007101731339A CN101221179A (en) | 2007-12-26 | 2007-12-26 | Bladder cancer detecting test paper |
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| Publication Number | Publication Date |
|---|---|
| CN101221179A true CN101221179A (en) | 2008-07-16 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2007101731339A Pending CN101221179A (en) | 2007-12-26 | 2007-12-26 | Bladder cancer detecting test paper |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101718792A (en) * | 2009-12-10 | 2010-06-02 | 兰州大学 | Protein chip for diagnosing bladder tumors |
| CN101974087A (en) * | 2009-03-31 | 2011-02-16 | 宝生物工程株式会社 | Anti-fibronectin fragment monoclonal antibody |
-
2007
- 2007-12-26 CN CNA2007101731339A patent/CN101221179A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974087A (en) * | 2009-03-31 | 2011-02-16 | 宝生物工程株式会社 | Anti-fibronectin fragment monoclonal antibody |
| CN101718792A (en) * | 2009-12-10 | 2010-06-02 | 兰州大学 | Protein chip for diagnosing bladder tumors |
| CN101718792B (en) * | 2009-12-10 | 2013-09-18 | 兰州大学 | Protein chip for diagnosing bladder tumors and preparation method thereof |
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Application publication date: 20080716 |