CN202330433U - Rapid test paper for acidophilic thiobacillus ferrooxidans - Google Patents
Rapid test paper for acidophilic thiobacillus ferrooxidans Download PDFInfo
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- CN202330433U CN202330433U CN2011204507727U CN201120450772U CN202330433U CN 202330433 U CN202330433 U CN 202330433U CN 2011204507727 U CN2011204507727 U CN 2011204507727U CN 201120450772 U CN201120450772 U CN 201120450772U CN 202330433 U CN202330433 U CN 202330433U
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- test paper
- band
- liking
- acid oxidase
- quick detection
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Abstract
The utility model discloses rapid test paper for acidophilic thiobacillus ferrooxidans and relates to the bacterial test field. The test paper comprises a plastic baseboard, a sample adding area, a calibration area, a developer area and a water suction area, wherein the sample adding area is a micro filtration membrane with a certain aperture; the calibration area is a glass cellulose membrane combined with A.f polyclonal antibodies which are marked by colloid selenium nanoparticles; a reaction zone consists of a nitrocellulose membrane with a test band (T band) and a quality control band (C band), wherein the detection band is embedded with A.f polyclonal antibodies, and the quality control band is combined with goat-anti-mouse IgG. The test paper has the advantages of being simple in operation, peculiar in sensitivity, convenient to carry, and capable of being used in A.f test of various circulating cooling water, oil extraction wastewater, pipe water and natural water.
Description
Technical field
The utility model relates to the field of Bacteria Detection, relates to a kind of ferrous Thiobacillus quick detection test paper of acid oxidase of having a liking for specifically.
Background technology
Have a liking for the ferrous Thiobacillus of acid oxidase (
Acidithiobacillus ferrooxidans, A.f) be a kind of chemautotrophy, obligate aerobic, have a liking for sour Gram-negative bacteria, mainly be grown in the environment of pH 1.0-4.5, through Oxidation of Fe
2+Obtain the required energy of growth course with lower valency sulphur.In various water deliverys, oil transporting appliance, cool cycles water pipe, producing well, heating installation return pipe and municipal drainage pipeline, all finding has
A.fExist, because
A.fCan the ferrous high ferro oxidate that is oxidized in the water be formed filemot dross in the thalline epitheca or on every side in the rete malpighii in the metal surface, oxygen gets into difficulty and causes oxygen-concentration cell corrosion in the dross, and then quickens corrosive pipeline.In addition,
A.fCan also the oxidation of the elementary sulfur in the water body and reductibility sulfuration thing finally be produced sulfuric acid, thus corroding metal, concrete component etc.
A.fCorrosion behavior all caused loss in various degree for commercial unit and civilian installation, the suitable heaviness of the economic loss aspect petrochemical complex and power-equipment especially.Therefore people also more and more pay close attention to its effective detection method.
At present,
A.fThe detection method that whether exists mainly contains the oxidation susceptibility method that detects ferrous ion after the enrichment of cultivating, multitube fermentation method, test bottle method, based on the detection 16S rRNA gene and the ferrous oxidase gene approach of Protocols in Molecular Biology.These methods all need be in the sample
A.fCarry out enrichment culture, the configuration of nutrient culture media is complicated, running program is loaded down with trivial details, detection time is long, high to test environment and operating personnel's competency profiling.In recent years, the Protocols in Molecular Biology that does not rely on cultivation also begin to use with
A.fDetection, according to 16S rRNA gene design fluorescence probe, come in the judgement sample through molecular hyridization then
A.fContain that there is something special.In addition, (Rusticyanin, Rus) the PCR method that contains two pairs of primers of encoding gene rus foundation also is applied to according to CER
A.fDetection.Though these two kinds of methods have certain sensitivity and specificity, the preparation of its reagent is complicated, to instrument requirement is high, testing cost is high and need special technician to accomplish.
Summary of the invention
The purpose of the utility model be to provide a kind of have high specific, high sensitivity, detection speed fast, need not professional's operation, cost and characteristics such as be easy to carry have a liking for the ferrous Thiobacillus quick detection test paper of acid oxidase.
The utility model have a liking for the ferrous Thiobacillus quick detection test paper of acid oxidase, be on plastic base, to have encapsulated sample application zone, calibration zone, reaction solution district, and suction zones, the above-mentioned overlapping successively overlap joint of belt edge of respectively distinguishing successively.
As the further improvement of the utility model, described sample application zone is made up of miillpore filter.
As the further improvement of the utility model, described calibration zone is made up of the plain film of spun glass, is coated with electroselenium particle mark on this film
A.fPolyclonal antibody.
As the further improvement of the utility model, described reaction solution district is made up of nitrocellulose membrane, and the reaction solution district is provided with detects band and quality control band.
As the further improvement of the utility model, be embedded with at the detection band
A.fPolyclonal antibody, quality control band are combined with sheep anti-mouse igg.
As the further improvement of the utility model, described suction zones is made up of filter paper.
The utility model have a liking for the ferrous Thiobacillus quick detection test paper of acid oxidase, this test paper utilizes
A.fPolyclonal antibody detects in the sample to be checked
A.fThereby, improved the rapidity, the sensitivity that detect.Utilize the colloidal nano granules of selenium mark
A.fPolyclonal antibody has improved the visuality of testing result, has improved the visuality of testing result; Further improved sensitivity, the preparation of colloidal nano granules of selenium requires low than the colloidal nano gold in addition, thereby has reduced this test strips necessary requirement condition of preparation; Widen test strips validity and detected required condition and range; Broken away from the constraint of particular instrument, reduced cost, made that the use object of this test strips is accurate, convenient, cheap, popular more.
Description of drawings
Fig. 1 is for having a liking for the ferrous Thiobacillus quick detection test paper of acid oxidase structural representation;
Fig. 2 is the exploded view of Fig. 1;
Fig. 3 is this test strips testing result key diagram.
Embodiment
The utility model have a liking for the ferrous Thiobacillus quick detection test paper of acid oxidase, be on plastic base 1, to have encapsulated sample application zone 2, calibration zone 3, reaction solution district 4, and suction zones 7, the above-mentioned overlapping successively overlap joint of belt edge of respectively distinguishing successively.
Described sample application zone 2 is made up of miillpore filter; Calibration zone 3 is made up of the plain film of spun glass, is coated with electroselenium particle mark on this film
A.fPolyclonal antibody; Reaction solution district 4 is made up of nitrocellulose membrane, reaction solution district 4 be provided with detect be with 5 with quality control band 6, be with 5 to be embedded with in detection
A.fPolyclonal antibody, quality control band 6 are combined with sheep anti-mouse igg; Suction zones 7 is made up of filter paper.
Have or not through detecting the colour developing situation of band, just can detecting in the sample convenient, fast, delicately
A.fExist.This kind method has facilitation, sensitization, specialization, in good timeization, characteristics such as rapid, popular, is fit to sample is carried out fast detecting.
The preparation process of having a liking for the ferrous Thiobacillus quick detection test paper of acid oxidase of the utility model is following:
1, the preparation of antigen
A.f(be made up of solution A and solution B, solution A comprises: 3.0 g (NH4) at 9 K nutrient culture media
2SO
4, 0.1 g KCl, 0.5 g K
2HPO
4, 0.5g MgSO
4.7H
2O, 0.01 g Ca (NO
3)
2, 700 mL distilled water; Solution B is that concentration is 14.78% FeSO
4Solution 300 mL.A liquid and B liquid mix and use rare H during use
2SO
4Transfer about pH to 2.0) in 30
oC cultivates, and centrifugal 15 min of nutrient solution 2000 rpm of the phase of taking the logarithm remove ferrovanadium, centrifugal 20 min of supernatant 8000 rpm, and deposition is with rare H of pH 2.0
2SO
4Washing is not to there being ferrovanadium, and the PBS with 0.02 M pH 7.4 cleans thalline to remove the acid solution on thalline surface then, gets deposition with 0.25 M NaCl washed twice, 4
oC is suspended in thalline in the NaCl solution of 1 mL-20 at last with centrifugal 20 min of l2000 rpm
oC preserves subsequent use.
2, the preparation of colloidal nano granules of selenium
(1) deionized water of adding a certain amount of (10 mL) in the beaker of cleaning, and with the lasting stirring of magnetic stirring apparatus middling speed;
(2) shitosan 0.2 mL that adds finite concentration (5.0%) is to stablize its reaction system;
Behind (3) 2 min, add 0.2 mL ascorbic acid and 0.1 mL selenous acid successively;
(4) treat that it reacts 5 min after, 1 min slowly drips 1 mL selenous acid and 2 mL ascorbic acid at interval successively.After adding, continue stirring reaction 5 min, finally process satisfactory colloidal nano granules of selenium.
3, electroselenium mark
A.fPolyclonal antibody
(1) confirms the electroselenium mark through preliminary experiment
A.fThe optimal pH of polyclonal antibody and concentration;
(2) with the electroselenium solution that makes through be adjusted to optimal pH (pH 8.2) with 0.1 M sal tartari;
(3) will
A.fPolyclonal antibody is removed impurity such as some ions that influence selenium mark process and particle as far as possible through processing such as further dialysis, filtrations;
(4) under the electromagnetic agitation condition, with treated
A.fPolyclonal antibody dropwise joins in the electroselenium solution according to optimizing the condition concentration that;
Behind (5) 10 min, add 10% BSA to its concentration be 1%, to stablize its reaction system, continue to stir;
Behind (6) 10 min, with the selenium mark that makes
A.fPolyclonal antibody solution concentrates through gradient centrifugation or dialysis concentrates through sucrose, obtains the selenium mark of high concentration degree
A.fPolyclonal antibody solution gets final product through gel chromatography column purifying, TBS (1% BSA, 0.05% Sodium azide) it again after washing.
4, have a liking for the assembling of the ferrous Thiobacillus quick detection test paper of acid oxidase
(1) with spun glass through after containing PBS (pH 7.4, the 0.01 M) activation processing of l% BSA and 1% Tween-20, be immersed in the selenium mark
A.f40 min in the polyclonal antibody solution, natural air drying can be marked selenium
A.fPolyclonal antibody is coated on the glass fibre membrane;
(2) spray successively at interval 0.4 cm on the nitrocellulose filter
A.fPolyclonal antibody solution band is as detecting band, and sheep anti-mouse igg is a quality control band.Then nitrocellulose filter is sealed 2 h, natural air drying with the PBS (pH 7.4,0.01 M) that contains l% BSA;
(3) last, plastic bottom board, with application of sample film, the plain film of spun glass, nitrocellulose filter and thieving paper to specifications the der group of accompanying drawing 1, Fig. 2 install, it is subsequent use to keep dry.
The use of test strips
(1) sample to be tested is splashed into sample application zone, or test strips stretched into to begin in the sample to detect.Get final product observations behind the 5-10 min.
(2) result judges (combining Figure of description 3)
All develop the color if detect band and quality control band, then interpret sample is positive, and contains in the sample
A.f, shown among Fig. 31; Do not develop the color if detect band, and the quality control band colour developing, then interpret sample is negative, and does not have in the sample
A.fExist, shown among Fig. 32; If detection band and detection band all do not develop the color, explain that then this test strips lost efficacy, can not use, detect again after the test strips that must more renew, shown among Fig. 33.
Claims (6)
1. have a liking for the ferrous Thiobacillus quick detection test paper of acid oxidase, it is characterized in that on plastic base (1), having encapsulated successively sample application zone (2), calibration zone (3), reaction solution district (4), and suction zones (7), the above-mentioned overlapping successively overlap joint of belt edge of respectively distinguishing.
2. as claimed in claim 1ly have a liking for the ferrous Thiobacillus quick detection test paper of acid oxidase, it is characterized in that sample application zone (2) is made up of miillpore filter.
3. as claimed in claim 1ly have a liking for the ferrous Thiobacillus quick detection test paper of acid oxidase, it is characterized in that calibration zone (3) is made up of the plain film of spun glass, be coated with electroselenium particle mark on this film
A.fPolyclonal antibody.
4. as claimed in claim 1ly have a liking for the ferrous Thiobacillus quick detection test paper of acid oxidase, it is characterized in that reaction solution district (4) is made up of nitrocellulose membrane, reaction solution district (4) are provided with and detect band (5) and quality control band (6).
5. as claimed in claim 4ly have a liking for the ferrous Thiobacillus quick detection test paper of acid oxidase, it is characterized in that detecting band (5) and be embedded with
A.fPolyclonal antibody, quality control band (6) are combined with sheep anti-mouse igg.
6. as claimed in claim 1ly have a liking for the ferrous Thiobacillus quick detection test paper of acid oxidase, it is characterized in that suction zones (7) is made up of filter paper.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011204507727U CN202330433U (en) | 2011-11-15 | 2011-11-15 | Rapid test paper for acidophilic thiobacillus ferrooxidans |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011204507727U CN202330433U (en) | 2011-11-15 | 2011-11-15 | Rapid test paper for acidophilic thiobacillus ferrooxidans |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN202330433U true CN202330433U (en) | 2012-07-11 |
Family
ID=46442409
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011204507727U Expired - Fee Related CN202330433U (en) | 2011-11-15 | 2011-11-15 | Rapid test paper for acidophilic thiobacillus ferrooxidans |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN202330433U (en) |
-
2011
- 2011-11-15 CN CN2011204507727U patent/CN202330433U/en not_active Expired - Fee Related
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120711 Termination date: 20121115 |