CN101661037A - Spittle rapid detection strip for chlamydia trachomatis antibody - Google Patents
Spittle rapid detection strip for chlamydia trachomatis antibody Download PDFInfo
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- CN101661037A CN101661037A CN200910152614A CN200910152614A CN101661037A CN 101661037 A CN101661037 A CN 101661037A CN 200910152614 A CN200910152614 A CN 200910152614A CN 200910152614 A CN200910152614 A CN 200910152614A CN 101661037 A CN101661037 A CN 101661037A
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- chlamydia trachomatis
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Abstract
The invention discloses a spittle rapid detection strip for a chlamydia trachomatis antibody, comprising a baseboard, to which a sample pad, a colloidal gold pad, a nitrocellulose membrane and a sample-drawing pad are sequentially adhered, wherein a mouse-anti-human IgG antibody which is marked by the colloidal gold and can be integrated with the specificity of a chlamydia trachomatis antibody isattached to the colloidal gold pad; and the nitrocellulose membrane is coated with a detection line formed by the chlamydia trachomatis antibody and a quality control line formed by the goat-anti-mouse antiantibody which can be integrated with the specificity of mouse-anti-human IgG antibody. The spittle rapid detection strip provided by the invention adopts the colloidal gold marking technology to detect the chlamydia trachomatis antibody in spittle so as to determine whether the person is infected with chlamydia trachomatis, and has the advantages that operation is simple; reaction is quick;sensitivity is high; specificity is strong; and the spittle rapid detection strip can be applied to both on-site detection and self-detection, and is economical and practical and the like.
Description
Technical field
The present invention relates to the biological diagnosis technical field, relate in particular to a kind of Spittle rapid detection strip for chlamydia trachomatis antibody.
Background technology
The record of " Bai Shi system handbook " (1984) is divided into 3 bions with chlamydia trachomatis Chlamydiatrachomatis, i.e. mouse bion (biovar mouse), trachoma bion (biovar trachoma) and lymphogranuloma venereum bion.The latter two are relevant with human diseases.Use indirect microimmunofluorescence test, the trachoma bion is divided A, B, Ba, C, D, Da, E, F, G, H, I, Ia, J, a K14 serotype again, and the LGV bion has L1, L2, L2a, a L34 serotype again.
Urogenital infections is caused by trachoma biovariety D～K serotype through transmission through sex.The male sex shows as urethritis more, can alleviate without treatment, but that majority is transformed into is chronic, periodically increases the weight of, and can merge epididymis inflammation, rectitis etc.The women can cause urethritis, cervicitis etc., and salpingitis is than severe complication.This serotype also can cause chlamydia trachomatis property pneumonia sometimes.
Lymphogranuloma venereum is caused by chlamydia trachomatis LGV biovariety.LGV will pass through two transmissions through sex, is a kind of venereal disease.The male sex invades inguinal lymph nodes, causes purulent lymphadenitis and chronic lymphatic granuloma.The women can invade perineum, anus, rectum, and it is narrow perineum～anus～rectal tissue to occur.
At present, detecting the most frequently used method of above-mentioned infectious disease is immune detection, and the method comprises two kinds of traditional enzyme linked immunosorbent assay (ELISA) and the recent colloidal gold methods that rises based on the specific recognition of antigen one antibody.Colloidal gold method is divided into immunochromatography and immunity percolation dual mode, and is wherein convenient, fast with immunochromatographic method.
The immunochromatography colloidal gold technique is novel diagnostic techniques, obtained using comparatively widely, ultimate principle is as follows: utilize a kind of antigen of colloid gold label or antibody, bag is matched antigen or antibody accordingly on the nitrocellulose filter of test strips, during detection when containing corresponding specific antibody or antigen in the sample, the part formation compound that combines in colloid gold label particle and the sample, chromatography on nitrocellulose filter then, coated again antigen or antibody capture, form macroscopic detection line (T line), have or not the judgement of realization the result by detection line.But existing test strips is a sample with blood mostly, is not suitable for on-the-spot the detection and self check, and it is higher to detect cost.
Summary of the invention
The invention provides a kind of Spittle rapid detection strip for chlamydia trachomatis antibody, this test strips can solve existing test strips and be not suitable for on-the-spot the detection with saliva as sample, detects the cost problem of higher.
A kind of Spittle rapid detection strip for chlamydia trachomatis antibody, comprise base plate, be pasted with sample pad, collaurum pad, nitrocellulose filter on the base plate successively and inhale the sample pad, be attached with the mouse-anti human IgG antibody that can combine of colloid gold label on the described collaurum pad, be coated with detection line that constitutes by trachoma chlamydia antigen and the nature controlling line that constitutes by sheep anti mouse two antiantibodys that can combine on the described nitrocellulose filter with mouse-anti human IgG antibody specificity with the chlamydia trachomatis antibody specificity.
Described chlamydia trachomatis antibody has 3 bions, that medically detect venereal disease mainly is poradenia bion (biovar lymphogranulomavenereum, LGV), the LGV bion has L1, L2, four serotypes of L2a, L3 again, chlamydia trachomatis antibody have 4 dissimilar, be medically to detect the label that whether infects chlamydia trachomatis, this label is present in the blood of human body in a large number, and a small amount of existence is also arranged in people's saliva.Chlamydia trachomatis has common antigen and is present in the former coating of clothing, and be present in chlamydial whole life cycle, described trachoma chlamydia antigen is exactly this common antigen, and is a kind of people's recombinant antigen, can combine with all serotype chlamydia trachomatis antibodies of LGV bion.Described mouse-anti human IgG antibody, trachoma chlamydia antigen and mouse-anti human IgG antibody all are commercially available prod, also can prepare by existing method.
Be attached with 0.18～0.30 μ g/cm on the described collaurum pad
2The mouse-anti human IgG antibody that can combine with the chlamydia trachomatis antibody specificity.It is the sensitivity requirement that sample detects with the saliva that this scope satisfies test strips of the present invention.
Described collaurum pad preparation method is as follows:
Get the colloidal gold solution that the colloid diameter is 15～35nm, regulate pH value to 8.0～8.2, in every 100ml colloidal gold solution, the mouse-anti human IgG antibody that can combine who adds 0.75～1.3mg with the chlamydia trachomatis antibody specificity, stirring the back seals with bovine serum albumin, the centrifugal supernatant that goes, the colloidal gold solution that adds equal volume redissolves, and every ml soln evenly is layered on 40cm
2On the glass fibre membrane, make the collaurum pad after the drying.
The content of trachoma chlamydia antigen is 0.08～1.4 μ g/cm in the described detection line; The sheep anti mouse two antiantibody content that can combine with described mouse-anti human IgG antibody specificity in the described nature controlling line are 0.08～1.4 μ g/cm.It is the sensitivity requirement that sample detects with the saliva that this scope satisfies test strips of the present invention.
Described detection line method for coating is as follows:
Described trachoma chlamydia antigen is dissolved in to make concentration in the phosphate buffer solution be 0.08～1.0mg/ml solution, with the end line of the consumption of 1～1.5 μ l/cm, promptly gets detection line after the drying at nitrocellulose filter with this solution;
Described nature controlling line method for coating is as follows:
Can be dissolved in sheep anti mouse two antiantibodys that described mouse-anti human IgG antibody specificity combines and make concentration in the phosphate buffer solution is 0.08～1.0mg/ml solution, consumption with this solution 1～1.5 μ l/cm is rule at the other end of nitrocellulose filter, promptly gets nature controlling line after the drying.
The present invention also provides a kind of detection method of above-mentioned Spittle rapid detection strip for chlamydia trachomatis antibody, may further comprise the steps:
Get people's saliva, with the phosphate buffer solution dilution, the saliva of getting after 50～100 μ l dilute drops on the sample pad, according to the color status of detection line (T line) and nature controlling line (C line), judges testing result.
All occur as C, T line, judge that sample is positive; The C line occurs, and the T line does not occur, and judges that sample is negative; C, T line do not occur or the appearance of T line does not appear in the C line, judge that all test paper is invalid.
Test strips of the present invention adopts colloidal gold-labeled method, detect the chlamydia trachomatis antibody in the saliva, thereby judge whether to infect chlamydia trachomatis, have simple to operate, reaction fast, high, the high specificity of susceptibility, be fit to on-the-spot detection and self check, advantage such as economical and practical.
Embodiment
Embodiment 1
The preparation of collaurum pad
Prepare the colloidal gold solution of diameter 15nm with gold chloride-trisodium citrate reduction method, get the 100ml colloidal gold solution in beaker after preparation is finished, use 0.1M K
2CO
3Transfer to pH8.0, add the mouse-anti human IgG antibody (Shanghai Linc-Bio Science Co., Ltd.) that 0.75mg can combine with the chlamydia trachomatis antibody specificity, stirring at room 1 hour, added percentage by weight and be 5% bovine serum albumin(BSA) (BSA) sealing 1 hour, 12000rpm, 4 ℃ are centrifugal 30 minutes, abandon supernatant, redissolve to 100ml, press 1ml solution shop 40cm with colloidal gold solution
2Ratio, solution is layered on the glass fibre membrane equably, 37 ℃ of dryings 30 minutes are made the collaurum pad.
The bag quilt of nitrocellulose filter
The trachoma chlamydia antigen (Shanghai Linc-Bio Science Co., Ltd.) that can combine with the chlamydia trachomatis antibody specificity is dissolved in the solution that 0.01M, pH8.0 phosphate buffer (PBS) are mixed with 0.08mg/ml, rules with the parameter of 1 μ l/cm at nitrocellulose filter one end with spray film instrument.
Can be dissolved in 0.01M, pH8.0 phosphate buffer (PBS) with sheep anti mouse two antiantibodys (Shanghai Linc-Bio Science Co., Ltd.) that above-mentioned mouse-anti human IgG antibody specificity combines and be mixed with the solution of 0.08mg/ml, rule with the parameter of 1 μ l/cm at the nitrocellulose filter other end with spray film instrument.After the line at room temperature dry 8 hours, obtain detection line and nature controlling line respectively.
Detect the assembling of test paper
In hothouse, 20～25 ℃ of temperature, humidity is less than 30%, sample pad (glass fibre membrane), collaurum pad, nitrocellulose filter and suction sample pad (suction test paper) are sticked on the base plate, wherein nitrocellulose filter is positioned at the base plate middle part, nitrocellulose filter is coated with an end of T line and 1/3 overlap joint of collaurum pad, is coated with an end and 1/10 overlap joint of inhaling the sample pad of C line, 1/5 overlap joint of sample pad and collaurum pad.Be cut into the test strips that width is 2.5mm at last, make test card in the plastic clip of also test strips can being packed into.
Embodiment 2
The preparation of collaurum pad
Prepare the colloidal gold solution of diameter 35nm with gold chloride-trisodium citrate reduction method, get the 100ml colloidal gold solution in beaker after preparation is finished, use 0.1M K
2CO
3Transfer to pH8.2, add the mouse-anti human IgG antibody (Shanghai Linc-Bio Science Co., Ltd.) that 1.3mg can combine with the chlamydia trachomatis antibody specificity, stirring at room 1 hour, added percentage by weight and be 5% bovine serum albumin(BSA) (BSA) sealing 1 hour, 12000rpm, 4 ℃ are centrifugal 30 minutes, abandon supernatant, redissolve to 100ml, press 1ml solution shop 40cm with colloidal gold solution
2Ratio, solution is layered on the glass fibre membrane equably, 37 ℃ of dryings 30 minutes are made the collaurum pad.
The bag quilt of nitrocellulose filter
The trachoma chlamydia antigen (Shanghai Linc-Bio Science Co., Ltd.) that can combine with the chlamydia trachomatis antibody specificity is dissolved in the solution that 0.01M, pH8.0 phosphate buffer (PBS) are mixed with 1.0mg/ml, rules with the parameter of 1.5 μ l/cm at nitrocellulose filter one end with spray film instrument.
Can be dissolved in 0.01M, pH8.0 phosphate buffer (PBS) with sheep anti mouse two antiantibodys (Shanghai Linc-Bio Science Co., Ltd.) that above-mentioned mouse-anti human IgG antibody specificity combines and be mixed with the solution of 1.0mg/ml, rule with the parameter of 1.5 μ l/cm at the nitrocellulose filter other end with spray film instrument.After the line at room temperature dry 8 hours, obtain detection line and nature controlling line respectively.
Detect the assembling of test paper
In hothouse, 20～25 ℃ of temperature, humidity is less than 30%, sample pad (glass fibre membrane), collaurum pad, nitrocellulose filter and suction sample pad (suction test paper) are sticked on the base plate, wherein nitrocellulose filter is positioned at the base plate middle part, nitrocellulose filter is coated with an end of T line and 1/3 overlap joint of collaurum pad, is coated with an end and 1/10 overlap joint of inhaling the sample pad of C line, 1/5 overlap joint of sample pad and collaurum pad.Be cut into the test strips that width is 2.5mm at last, make test card in the plastic clip of also test strips can being packed into.
The clinical sample test
People to be checked is gathering saliva sample fasting in preceding 30 minutes, get every people's saliva 0.5ml to be checked in dixie cup, get in two phosphate buffer solutions that are added drop-wise to concentration 0.02M, pH8.0 with suction pipe and to dilute, the saliva 100 μ l that get dilution are added in the sample pad, naked-eye observation 30 minutes, the color status of record T line and C line.Simultaneously saliva sample is detected with chlamydia trachomatis antibody ELISA kit, if both test result are inconsistent, detect with other two kinds of chlamydia trachomatis antibody ELISA kits again, if two kinds or above ELISA reagent are positive, result of determination is positive, two kinds or above ELISA reagent are negative, and result of determination is negative.
Collect 163 parts of saliva samples, wherein the ELISA kit detects 43 parts of positive, and test strips of the present invention (embodiment 1) detects 44 parts of positive, and concrete outcome is as shown in table 1 below:
Table 1 test strips of the present invention is to the testing result of clinical sample chlamydia trachomatis antibody
Sensitivity=48/48=100%; Specificity=116/117=99.1%.
Claims (6)
1, a kind of Spittle rapid detection strip for chlamydia trachomatis antibody, comprise base plate, be pasted with sample pad, collaurum pad, nitrocellulose filter on the base plate successively and inhale the sample pad, it is characterized in that: be attached with the mouse-anti human IgG antibody that can combine of colloid gold label on the described collaurum pad, be coated with detection line that constitutes by trachoma chlamydia antigen and the nature controlling line that constitutes by sheep anti mouse two antiantibodys that can combine on the described nitrocellulose filter with mouse-anti human IgG antibody specificity with the chlamydia trachomatis antibody specificity.
2, Spittle rapid detection strip for chlamydia trachomatis antibody according to claim 1 is characterized in that: be attached with 0.18～0.30 μ g/cm on the described collaurum pad
2The mouse-anti human IgG antibody that can combine with the chlamydia trachomatis antibody specificity.
3, Spittle rapid detection strip for chlamydia trachomatis antibody according to claim 1 and 2 is characterized in that, described collaurum pad preparation method is as follows:
Get the colloidal gold solution that the colloid diameter is 15～35nm, regulate pH value to 8.0～8.2, in every 100ml colloidal gold solution, the mouse-anti human IgG antibody that can combine who adds 0.75～1.3mg with the chlamydia trachomatis antibody specificity, stirring the back seals with bovine serum albumin, the centrifugal supernatant that goes, the colloidal gold solution that adds equal volume redissolves, and every ml soln evenly is layered on 40cm
2On the glass fibre membrane, make the collaurum pad after the drying.
4, Spittle rapid detection strip for chlamydia trachomatis antibody according to claim 1 is characterized in that: the content of trachoma chlamydia antigen is 0.08～1.4 μ g/cm in the described detection line; The sheep anti mouse two antiantibody content that can combine with described mouse-anti human IgG antibody specificity in the described nature controlling line are 0.08～1.4 μ g/cm.
According to claim 1 or 4 described Spittle rapid detection strip for chlamydia trachomatis antibody, it is characterized in that 5, described detection line method for coating is as follows:
Described trachoma chlamydia antigen is dissolved in to make concentration in the phosphate buffer solution be 0.08～1.0mg/ml solution, with the end line of the consumption of 1～1.5 μ l/cm, promptly gets detection line after the drying at nitrocellulose filter with this solution;
Described nature controlling line method for coating is as follows:
Can be dissolved in sheep anti mouse two antiantibodys that described mouse-anti human IgG antibody specificity combines and make concentration in the phosphate buffer solution is 0.08～1.0mg/ml solution, consumption with this solution 1～1.5 μ l/cm is rule at the other end of nitrocellulose filter, promptly gets nature controlling line after the drying.
6, the detection method of the described Spittle rapid detection strip for chlamydia trachomatis antibody of a kind of claim 1 may further comprise the steps:
Get people's saliva, with the phosphate buffer solution dilution, the saliva of getting after 50～100 μ l dilute drops on the sample pad, according to the color status of detection line and nature controlling line, judges testing result.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910152614A CN101661037A (en) | 2009-09-10 | 2009-09-10 | Spittle rapid detection strip for chlamydia trachomatis antibody |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910152614A CN101661037A (en) | 2009-09-10 | 2009-09-10 | Spittle rapid detection strip for chlamydia trachomatis antibody |
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| Publication Number | Publication Date |
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| CN101661037A true CN101661037A (en) | 2010-03-03 |
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|---|---|---|---|
| CN200910152614A Pending CN101661037A (en) | 2009-09-10 | 2009-09-10 | Spittle rapid detection strip for chlamydia trachomatis antibody |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102013192A (en) * | 2010-06-29 | 2011-04-13 | 上海杰人信息科技有限公司 | Spit judgment device and judgment method thereof |
| CN107884574A (en) * | 2017-11-01 | 2018-04-06 | 上海凯创生物技术有限公司 | Chlamydia trachomatis detection reagent card, kit and application thereof |
| CN112920270A (en) * | 2021-02-09 | 2021-06-08 | 杭州隆基生物技术有限公司 | Anti-chlamydia trachomatis recombinant antibody and application thereof |
| CN114414798A (en) * | 2021-12-08 | 2022-04-29 | 北京泰格科信生物科技有限公司 | Chlamydia trachomatis/gonococcus/mycoplasma genitalium antigen combined detection kit and preparation method thereof |
-
2009
- 2009-09-10 CN CN200910152614A patent/CN101661037A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102013192A (en) * | 2010-06-29 | 2011-04-13 | 上海杰人信息科技有限公司 | Spit judgment device and judgment method thereof |
| CN102013192B (en) * | 2010-06-29 | 2012-10-03 | 上海杰人信息科技有限公司 | Spit judgment device and judgment method thereof |
| CN107884574A (en) * | 2017-11-01 | 2018-04-06 | 上海凯创生物技术有限公司 | Chlamydia trachomatis detection reagent card, kit and application thereof |
| CN112920270A (en) * | 2021-02-09 | 2021-06-08 | 杭州隆基生物技术有限公司 | Anti-chlamydia trachomatis recombinant antibody and application thereof |
| CN112920270B (en) * | 2021-02-09 | 2023-01-10 | 杭州隆基生物技术有限公司 | Anti-chlamydia trachomatis recombinant antibody and application thereof |
| CN114414798A (en) * | 2021-12-08 | 2022-04-29 | 北京泰格科信生物科技有限公司 | Chlamydia trachomatis/gonococcus/mycoplasma genitalium antigen combined detection kit and preparation method thereof |
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Application publication date: 20100303 |