CN1866021A - Test paper for quick detection of cholera vibrio 01 group, 0139 group and cholera toxin - Google Patents
Test paper for quick detection of cholera vibrio 01 group, 0139 group and cholera toxin Download PDFInfo
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- CN1866021A CN1866021A CN 200610017786 CN200610017786A CN1866021A CN 1866021 A CN1866021 A CN 1866021A CN 200610017786 CN200610017786 CN 200610017786 CN 200610017786 A CN200610017786 A CN 200610017786A CN 1866021 A CN1866021 A CN 1866021A
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- vibrio cholerae
- monoclonal antibody
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Abstract
The related test paper to fast detect comma bacillus 01 group, 0139 group and cholera toxin comprises: base on PVC plate carrier, from bottom to top, connecting a sample pad, a colloidal gold conjunct to overlay and label single antibody for former comma bacillus and toxin, the cellulose nitrate covered by other three monoclonal antibody on T1, T2 and T3 areas and anti-rat IG on C area, and an absorption pad. This invention has accurate result, strong specificity, and can play important role in early diagnosis for cholera.
Description
One, technical field
The present invention relates to a kind of single job test strips of fast detecting vibrio cholerae 01 group, 0139 crowd and cholera toxin simultaneously.
Two, background technology
Cholera is a kind of acute infectious intestinal disease that the comma bacillus by Toxigenous commabacillus causes, because the cholera morbidity is anxious, propagation is fast, it is wide to involve scope, so classified as category A infectious disease by China's laws on contagious disease, there are seven cholera to be very popular on the human history, involve more than 100 country, captured up to ten million people's life, 01 sero-group is to cause these seven times pandemic main sero-groups.In November, 1992, a kind of comma bacillus novel, non-01 sero-group is found in India and Bangladesh country, be named as 0139 sero-group, 0139 group cholera vibrio feeds through to tens countries and regions very soon, predictedly causes the new world possibly be very popular behind 01 group cholera vibrio.Cholera toxin is a kind of protein by the comma bacillus secretion of causing a disease.It is made of two subunits of A, B, and wherein A subunit is the product poison part of cholera toxin, and it enters in the small intestine cells, effect by enzyme, change the secreting function of intestinal mucosa cells, cause body water and electrolytical a large amount of forfeiture, jeopardize people's life when serious.After all, producing malicious type comma bacillus is only and causes cholera outburst, popular arch-criminal.If after being separated to vibrio cholerae 01 type and 0139 type, must whether produce the detection of poison to the comma bacillus that separates, if do not produce cholera toxin, do not take anti-epidemic measure according to decree in effect.
In view of cholera brings human so serious threat, with the early stage diagnosis and treatment work of the cholera control emphasis as cholera, therefore, quick, simple and easy, sensitive, special, safe detection comma bacillus just seems particularly important to national correlation department.
At present to 01 group of cholera and 0139 group what make a definite diagnosis many employings is the isolation identification method of pathogen, though high specificity, but length consuming time, generally more than tens hours, be unfavorable in time making a definite diagnosis in early days, also have some conventional aided diagnosis methods as serum agglutination test, brake test etc., but there is poor specificity in these methods, problems such as sensitivity is low, also has a kind of highly sensitive method, be polymerase chain reaction (PCR), but make required in this way instrument costliness and complicated operation, also very easily, operational pollution occurs because of causing false positive, be not suitable for promoting the use of clinically, also there is report introduction to use the immunochromatographyassay assay comma bacillus at present, but what this method was used is the polyclonal antibody of anti-comma bacillus, and its specificity is just not as using the high specificity of monoclonal antibody, and the cross reaction of 01 group and 0139 group takes place easily.
The method of existing detection cholera toxin adopts polymerase chain reaction (PCR) more, its gene segment that produces poison is increased, though sensitivity is higher, but this method length consuming time, the expense height, strict to operating personnel's requirement, be unfavorable for the rapid screening of patient samples and popularizing of detection.
Three, summary of the invention
At above-mentioned situation, the present invention's purpose just provides a kind of 0139 crowd new of the fast detecting vibrio cholerae 01 group and the test strips of cholera toxin, can once solve fast and detect the vibrio cholerae 01 group, the multi-linked immunity chromatography problem of 0139 group and cholera toxin, the technical scheme of its solution is, this test strips is on PVC plate carrier, be connected with sample pad from bottom to top in turn, collaurum bond pad, nitrocellulose filter (being the N.C film) and absorption pad, nitrocellulose filter is mounted on above the PVC plate carrier, the mark that superposes respectively on the collaurum bond pad the anti-vibrio cholerae 01 group of strain monoclonal antibody, 39 groups of monoclonal antibodies of the anti-vibrio cholerae 01 of one strain and the anti-cholera toxin monoclonal antibody of a strain, or its three's mixing, T is arranged on the nitrocellulose filter
1District, T
2District, T
3District and C district, T
1District, T
2District, T
3Be coated with respectively in the district with collaurum bond pad on the anti-vibrio cholerae 01 group of the other strain monoclonal antibody that is complementary of monoclonal antibody, 39 groups of monoclonal antibodies of the anti-vibrio cholerae 01 of one strain and the anti-cholera toxin monoclonal antibody of a strain, the C district is coated with the antibody of anti-rat immune globulin, structure uniqueness of the present invention, easy to use, testing result is accurate, do not produce cross reaction with other enteric bacteria, high specificity, highly sensitive, fast, economical, safety, be specially adapted to the fast detecting and the evaluation of on-the-spot and clinical samples, the early diagnosis of cholera is had significant values and meaning.
Four, description of drawings
Fig. 1 is a structure side view of the present invention.
Fig. 2 is a structural front view of the present invention.
Five, embodiment
Below in conjunction with accompanying drawing the specific embodiment of the present invention is elaborated.
Provide by Fig. 1, Fig. 2, the present invention is on PVC plate carrier 1, be connected with sample pad 2, collaurum bond pad 3, nitrocellulose filter 4 and absorption pad 5 from bottom to top in turn, nitrocellulose membrane 4 is mounted on above the PVC plate carrier 1, the mark that superposes respectively on the collaurum bond pad 3 the anti-vibrio cholerae 01 group of strain monoclonal antibody, 39 groups of monoclonal antibodies of the anti-vibrio cholerae 01 of a strain and anti-cholera toxin monoclonal antibody of a strain or its three's mixing, on the nitrocellulose filter 4 T is arranged
1District, T
2District, T
3District and C district, T
1District, T
2District, T
3Be coated with respectively in the district with collaurum bond pad on the monoclonal antibody anti-vibrio cholerae 01 group of the other strain monoclonal antibody, 39 groups of monoclonal antibodies of the anti-vibrio cholerae 01 of a strain and the anti-cholera toxin monoclonal antibody of a strain that are complementary, the C district is coated with the antibody of anti-rat immune globulin.
In order to guarantee safe in utilization and result of use, the convex edge 7 that said nitrocellulose filter one end is absorbed pad 5 is fitted on the PVC plate carrier 1, and the other end is fitted on the PVC plate carrier 1 through the convex edge 6 of colloid bond pad 3 by sample pad 2.
Said anti-vibrio cholerae 01 group monoclonal antibody, 39 groups of monoclonal antibodies of anti-vibrio cholerae 01 and anti-cholera toxin monoclonal antibody are to adopt conventional current techique mouse spleen cell and myeloma cell's integration technology to prepare respectively, and by pairing to all monoclonal antibodies, screening, the surging monoclonal antibody of three couple (six strains) of a final selected antagonism vibrio cholerae 01 group, 39 groups of antagonism vibrio cholerae 01s and an antagonism cholera toxin, monoclonal antibody under the choosing through behind the conventional purification technique purifying, is placed collaurum bond and T respectively
1District, T
2District and T
3Get final product in the district.
Operating position of the present invention is:
A, directly get the about 150ul of patient specimen after patient's watery stool sample or basic peptone water increase bacterium, join the sample pad end of test strips with suction pipe.
Observations in B, 5-15 minute.
C, result judge: if having only the C district red stripes to occur, then negative; As in the C district, two red stripes appear in the T1 district, then are that 01 group cholera vibrio infects but do not produce poison; As in the C district and T1, T3 district three red stripes appear, then be that 01 group cholera vibrio infects and produces poison; As in the C district, two red stripes appear in the T2 district, then are that 0139 group cholera vibrio infects but do not produce poison; As in the C district and T2, T3 district three red stripes appear, then be that 0139 group cholera vibrio infects and produces poison.
In addition, in order to prevent to cause in the operating process pollution of pathogen, preferably again for this invention is equipped with test card, sample directly joins in the sample cell, can read the result in display window, because the test card good airproof performance is convenient to safe operation, also is convenient to after the use handle.
The biological starting material that the present invention uses are the anti-vibrio cholerae 01 group of two strains monoclonal antibody, 39 groups of monoclonal antibodies of the anti-vibrio cholerae 01 of two strains and the anti-cholera toxin monoclonal antibody of two strains of high specificity.
Through test, this test strips and the equal no cross reaction of following high concentration bacterium, these bacteriums comprise: Escherichia coli, shigella dysenteriae, vibrio parahemolyticus, vibrio mimicus, small intestine colon Yersinia ruckeri, Maxwell vibrios, Salmonella, staphylococcus aureus, typhoid bacillus, paratyphoid bacillus A, Bacillus paratyphosus B, Bacillus paratyphosus C, proteus, Shiga bacillus, Vibrio flurialis, citrobacter and Bifidobacterium Bifidum etc., test method are earlier above-mentioned bacterium to be diluted to 10 respectively
9Individual/ml, respectively get 150 μ l, be added drop-wise on the sample pad of test strips of the present invention, 5-15 minute, observations, all negative, show test strips of the present invention and above bacterium no cross reaction, high specificity, in addition, through 120 example tests, this test strips testing result accuracy rate is very high, accuracy rate is up to more than 98%, and through the evaluation of disease prevention and control center, Henan Province, the minimum recall rate that this test strips detects comma bacillus is 10
4-10
5Individual/ml, very outstanding have a high specificity, highly sensitive, fast, economy, the characteristics of safety, effectively overcome the existing detection comma bacillus time long, efficient is low, affects the problem of golden hour adversely, is specially adapted to the fast detecting of on-the-spot and clinical samples, solved that long-term people want to solve and the unresolved problem that quick and precisely detects comma bacillus, huge economic and social benefit has been arranged.
Claims (3)
1, the test strips of 0139 crowd of a kind of fast detecting vibrio cholerae 01 group and cholera toxin, it is characterized in that being, on PVC plate carrier (1), be connected with sample pad (2) from bottom to top in turn, collaurum bond pad (3), nitrocellulose filter (4) and absorption pad (5), nitrocellulose membrane (4) is mounted on above the PVC plate carrier (1), the mark that superposes respectively on the collaurum bond pad (3) the anti-vibrio cholerae 01 group of strain monoclonal antibody, 39 groups of monoclonal antibodies of the anti-vibrio cholerae 01 of one strain and anti-cholera toxin monoclonal antibody of a strain or its three's mixing, nitrocellulose filter has T on (4)
1District, T
2District, T
3District and C district, T
1District, T
2District, T
3Be coated with respectively in the district with collaurum bond pad on the monoclonal antibody anti-vibrio cholerae 01 group of the other strain monoclonal antibody, 39 groups of monoclonal antibodies of the anti-vibrio cholerae 01 of a strain and the anti-cholera toxin monoclonal antibody of a strain that are complementary, the C district is coated with the antibody of anti-rat immune globulin.
2, the test strips of 0139 crowd of fast detecting vibrio cholerae 01 group according to claim 1 and cholera toxin, it is characterized in that, the convex edge (7) that said nitrocellulose filter one end is absorbed pad (5) is fitted on the PVC plate carrier (1), and the other end is fitted on the PVC plate carrier (1) through the convex edge (6) of colloid bond pad (3) by sample pad (2).
3, the test strips of 0139 crowd of fast detecting vibrio cholerae 01 group according to claim 1 and cholera toxin, it is characterized in that, said anti-vibrio cholerae 01 group monoclonal antibody, 39 groups of monoclonal antibodies of anti-vibrio cholerae 01 and anti-cholera toxin monoclonal antibody are to adopt conventional current techique mouse spleen cell and myeloma cell's integration technology to prepare respectively, and by pairing to all monoclonal antibodies, screening, a final selected antagonism vibrio cholerae 01 group, the surging monoclonal antibody of three couples of 39 groups of one antagonism vibrio cholerae 01s and an antagonism cholera toxin, monoclonal antibody under the choosing through behind the conventional purification technique purifying, is placed collaurum bond and T respectively
1District, T
2District and T
3In the district.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610017786 CN1866021A (en) | 2006-05-16 | 2006-05-16 | Test paper for quick detection of cholera vibrio 01 group, 0139 group and cholera toxin |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610017786 CN1866021A (en) | 2006-05-16 | 2006-05-16 | Test paper for quick detection of cholera vibrio 01 group, 0139 group and cholera toxin |
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| CN 200610017786 Pending CN1866021A (en) | 2006-05-16 | 2006-05-16 | Test paper for quick detection of cholera vibrio 01 group, 0139 group and cholera toxin |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101000345B (en) * | 2006-11-28 | 2011-04-27 | 中国人民解放军军事医学科学院微生物流行病研究所 | Immunological chromatographic test paper for testing and its preparation method |
| CN102094090A (en) * | 2010-12-13 | 2011-06-15 | 华东师范大学 | Cholera toxin virulence gene detection kit and detection method thereof |
| CN102375059A (en) * | 2010-08-19 | 2012-03-14 | 中国疾病预防控制中心传染病预防控制所 | Cholera detecting and typing test paper based on multiple up-converting phosphor technology-based lateral-flow assays |
| CN102388309A (en) * | 2009-04-09 | 2012-03-21 | 日立化成工业株式会社 | Detector and detection method |
| CN104345150A (en) * | 2013-07-26 | 2015-02-11 | 深圳市艾瑞生物科技有限公司 | Glycated albumin detection immunochromatography test trip and preparation method thereof |
| CN106290842A (en) * | 2015-05-13 | 2017-01-04 | 上海凯创生物技术有限公司 | Cholera vibrio O 139 colloidal gold method detection kit |
| CN106290845A (en) * | 2015-05-13 | 2017-01-04 | 上海凯创生物技术有限公司 | Vibrio cholerae O 1 colloidal gold method detection kit |
-
2006
- 2006-05-16 CN CN 200610017786 patent/CN1866021A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101000345B (en) * | 2006-11-28 | 2011-04-27 | 中国人民解放军军事医学科学院微生物流行病研究所 | Immunological chromatographic test paper for testing and its preparation method |
| CN102388309A (en) * | 2009-04-09 | 2012-03-21 | 日立化成工业株式会社 | Detector and detection method |
| CN102375059A (en) * | 2010-08-19 | 2012-03-14 | 中国疾病预防控制中心传染病预防控制所 | Cholera detecting and typing test paper based on multiple up-converting phosphor technology-based lateral-flow assays |
| CN102375059B (en) * | 2010-08-19 | 2015-01-14 | 中国疾病预防控制中心传染病预防控制所 | Cholera detecting method |
| CN102094090A (en) * | 2010-12-13 | 2011-06-15 | 华东师范大学 | Cholera toxin virulence gene detection kit and detection method thereof |
| CN102094090B (en) * | 2010-12-13 | 2013-03-13 | 华东师范大学 | Cholera toxin virulence gene detection kit and detection method thereof |
| CN104345150A (en) * | 2013-07-26 | 2015-02-11 | 深圳市艾瑞生物科技有限公司 | Glycated albumin detection immunochromatography test trip and preparation method thereof |
| CN106290842A (en) * | 2015-05-13 | 2017-01-04 | 上海凯创生物技术有限公司 | Cholera vibrio O 139 colloidal gold method detection kit |
| CN106290845A (en) * | 2015-05-13 | 2017-01-04 | 上海凯创生物技术有限公司 | Vibrio cholerae O 1 colloidal gold method detection kit |
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