CN202330437U

CN202330437U - Colloidal gold test paper used for detecting acidithiobacillus ferrooxidans strains

Info

Publication number: CN202330437U
Application number: CN2011204372768U
Authority: CN
Inventors: 晏磊; 张爽
Original assignee: Heilongjiang Bayi Agricultural University
Current assignee: Heilongjiang Bayi Agricultural University
Priority date: 2011-11-08
Filing date: 2011-11-08
Publication date: 2012-07-11
Anticipated expiration: 2021-11-08

Abstract

The utility model relates to colloidal gold test paper used for detecting acidithiobacillus ferrooxidans strains. The test paper comprises a plastic base plate (1), a microfiltration membrane (2), a glass cellulose membrane (3), a nitrocellulose membrane (4) and filter paper (5), wherein the microfiltration membrane (2), the glass cellulose membrane (3), the nitrocellulose membrane (4) and the filter paper (5) are sequentially pasted on the plastic base plate (1); and an Rus monoclonal antibody I marked by colloidal gold particles is arranged on the glass cellulose membrane (3), and the nitrocellulose membrane (4) is provided with an Rus monoclonal antibody II stripe (6). According to the utility model, principles of antigen-antibody reaction and immunochromatography technology are utilized, the Rus monoclonal antibody is prepared onto the test paper, existence of Rus is detected by virtue of coloration of the colloidal gold particles by utilizing the characteristic that the Rus monoclonal antibody can be combined with the Rus, and whether A. f strains exist is judged further, and thus the test paper is convenient to use.

Description

The colloid gold test paper of the ferrous Thiobacillus of acid oxidase is had a liking in a kind of detection

Technical field

The utility model relates to the detection test paper, concrete is the colloid gold test paper that the ferrous Thiobacillus of acid oxidase is had a liking in a kind of detection.

Background technology

Have a liking for the ferrous Thiobacillus of acid oxidase ( Acidithiobacillus ferrooxidans, hereinafter to be referred as 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, with Oxidation of Fe ²⁺, S ⁰And S ^2-Compound wait and obtain the required energy of growth course.This bacterium is widely used in the recovery of biological metallurgy, useful metal, the desulfurization of coal and the processing of sulfur-containing waste water, is in the resource micro-organism treatment process, to use maximum, that adaptability is the strongest a kind of bacteriums.In water delivery, 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 be with elementary sulfur in the water body and the oxidation of reductibility sulfuration thing; The final sulfuric acid that produces; Thereby corroding metal, concrete component etc., its corrosion behavior bring serious harm for facilities such as underground utilities, subsea cable, oil production equipment, industrial waterflood system, cause loss economically.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, the PCR method of setting up according to CER (Rusticyanin is hereinafter to be referred as Rus) encoding gene rus that contains two pairs of primers also is applied to 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.

Rus is A.fA kind of important protein in the ferrous oxidation process, the albumen of each molecule comprise a copper atom, and molecular weight is 16.5 kDa.In the iron growing environment A.fThalline contains the Rus of high concentration, can reach 5% of total soluble protein.From difference A.fThe Rus that separates in the bacterial strain, its amino acid sequence homology is very high, and in other gramnegative bacterium, does not find to have the existence of Rus as yet, so Rus can be used as A.fOne well detect sign.Need a kind of Rus of utilization albumen of invention to detect A.fThe device that whether exists.

Summary of the invention

The goal of the invention of the utility model is the colloid gold test paper that provides a kind of detection to have a liking for the ferrous Thiobacillus of acid oxidase to the problems referred to above, solves at present and detects A.fThe method that whether exists is too complicated, the cost problem of higher.

The utility model is realized through following technical scheme: the colloid gold test paper of the ferrous Thiobacillus of acid oxidase is had a liking in a kind of detection; This test paper comprises plastic bottom board, miillpore filter, the plain film of spun glass, nitrocellulose filter, filter paper, posts miillpore filter, the plain film of spun glass, nitrocellulose filter and filter paper on the plastic bottom board successively; The Rus monoclonal antibody I of colloid gold particle mark is arranged on the plain film of described spun glass, Rus monoclonal antibody II band is arranged on the nitrocellulose filter.

Also have the sheep anti-mouse igg band on the above-mentioned nitrocellulose filter, Rus monoclonal antibody II band and sheep anti-mouse igg band laterally arrange, and Rus monoclonal antibody II band is between the Rus of colloid gold particle mark monoclonal antibody I and sheep anti-mouse igg band.

Adopt the good effect of technique scheme: the utility model utilizes the principle of antigen-antibody reaction and immunochromatography technique; With the Rus Monoclonal Antibody to test paper; Know from experience the characteristic that combines with Rus through the Rus monoclonal anti; Through the colloid gold particle colour developing, detect the existence of Rus, and then judge A.fWhether exist; Simultaneously, increased the sheep anti-mouse igg band, can detect the utility model delicately and whether lose efficacy, the convenient use.

Description of drawings

Fig. 1 is the vertical view of the utility model;

Fig. 2 is the assembling synoptic diagram of the utility model.

Among the figure: 1 plastic bottom board, 2 miillpore filters, the plain film of 3 spun glass, 4 nitrocellulose filters, 5 filter paper, 6 Rus monoclonal antibody II bands, 7 sheep anti-mouse igg bands.

Embodiment: the utility model is described further below in conjunction with accompanying drawing:

Fig. 1 is the vertical view of the utility model; Fig. 2 is the assembling synoptic diagram of the utility model; In conjunction with Fig. 1, shown in Figure 2; The colloid gold test paper of the ferrous Thiobacillus of acid oxidase is had a liking in a kind of detection, and this test paper comprises plastic bottom board 1, miillpore filter 2, the plain film 3 of spun glass, nitrocellulose filter 4, filter paper 5, posts miillpore filter 2, the plain film 3 of spun glass, nitrocellulose filter 4 and filter paper 5 on the plastic bottom board 1 successively.Miillpore filter 2 is used for application of sample, and filter paper 5 is mainly used in draws unnecessary liquid.The Rus monoclonal antibody I of colloid gold particle mark is arranged on the plain film 3 of spun glass, utilize antigen-antibody reaction, if contain Rus in the solution to be measured, the Rus monoclonal antibody I of Rus meeting and colloid gold particle mark combines.Rus monoclonal antibody II band 6 is arranged on the nitrocellulose filter 4; Because Rus monoclonal antibody II is sprayed into ribbon; Rus can continue to combine with Rus monoclonal antibody II with the compound that the Rus monoclonal antibody I of colloid gold particle mark combines the back to form like this; Thereby 6 places deposit at Rus monoclonal antibody II band, produce colour developing, thereby judge whether contain Rus in the solution to be measured.

Whether lost efficacy in order detection at any time to go out test paper; Judging out in the situation that Rus monoclonal antibody II band 6 does not develop the color is because do not contain Rus in the solution to be measured; Still the Rus monoclonal antibody I that causes the colloid gold particle mark disappears because test paper lost efficacy; On nitrocellulose filter 4, also be provided with sheep anti-mouse igg band 7; Rus monoclonal antibody II band 6 laterally arranges with sheep anti-mouse igg band 7, and Rus monoclonal antibody II band 6 is between the Rus of colloid gold particle mark monoclonal antibody I and sheep anti-mouse igg band 7.Because Rus monoclonal antibody I and Rus monoclonal antibody II all derive from mouse; Therefore sheep anti-mouse igg can mutually combine with Rus monoclonal antibody I and Rus monoclonal antibody II; Because Rus monoclonal antibody I also is marked with colloid gold particle; When the Rus monoclonal antibody I that is marked with colloid gold particle and sheep anti-mouse igg combine, can deposit at sheep anti-mouse igg band 7 places, thereby develop the color.On normal condition; A certain amount of Rus monoclonal antibody I that is marked with colloid gold particle is arranged on the test paper; No matter whether contain Rus in the testing liquid, the Rus monoclonal antibody I that is marked with colloid gold particle all can combine with sheep anti-mouse igg, thereby makes the colour developing of sheep anti-mouse igg band 7 places; If test paper had lost efficacy, when promptly not being marked with the Rus monoclonal antibody I of colloid gold particle, chromogenic reaction just can not take place in sheep anti-mouse igg band 7 yet, at this moment, reminds the experimenter to change test paper again and detects.

When this test strips is used for test sample; Miillpore filter 2 places of pretreated sample drop in this test strips; Sample is because capillary action is carried out lateral chromatography, through the plain film 3 of spun glass, if in the sample Rus is arranged; It can combine with the Rus monoclonal antibody I of colloid gold label, forms Ag-Ab I-colloidal gold composite.Continue then to ooze to move, combine, form Ab II-Ag-Ab I-colloidal gold composite, thereby deposit, be macroscopic band at Rus monoclonal antibody II band 6 places with Rus monoclonal antibody II fixing on the nitrocellulose filter 4.Continue then to ooze to move; Because Rus monoclonal antibody I and Rus monoclonal antibody II all derive from mouse; When running into sheep anti-mouse igg band 7, meeting and sheep anti-mouse igg combine, and the Rus monoclonal antibody I of colloid gold label presents macroscopic band at sheep anti-mouse igg band 7 places.

If do not have Rus in the sample, then can not form Ag-Ab I-colloidal gold composite, thereby can not combine with Rus monoclonal antibody II, Rus monoclonal antibody II band 6 does not develop the color.But, no matter in the sample whether Rus being arranged, the Rus monoclonal antibody I of colloid gold label all can be oozed under capillary action and moved on to sheep anti-mouse igg band 7 places, combines with sheep anti-mouse igg, makes 7 colour developings of sheep anti-mouse igg band.

In the detection,, prove in the solution to be measured and contain Rus, and then proved if Rus monoclonal antibody II band 6 all develops the color with sheep anti-mouse igg band 7 A.fExistence; If Rus monoclonal antibody II band 6 does not develop the color, 7 colour developings of sheep anti-mouse igg band prove and do not contain Rus in the solution to be measured promptly do not have A.fExistence; If Rus monoclonal antibody II band 6 does not all develop the color with sheep anti-mouse igg band 7, prove that test strips lost efficacy, need change test paper again and detect.

Claims

1. the colloid gold test paper of the ferrous Thiobacillus of acid oxidase is had a liking in a detection; It is characterized in that: this test paper comprises plastic bottom board (1), miillpore filter (2), the plain film (3) of spun glass, nitrocellulose filter (4), filter paper (5), posts miillpore filter (2), the plain film (3) of spun glass, nitrocellulose filter (4) and filter paper (5) on the plastic bottom board (1) successively; The Rus monoclonal antibody I of colloid gold particle mark is arranged on the plain film of described spun glass (3), Rus monoclonal antibody II band (6) is arranged on the nitrocellulose filter (4).

2. the colloid gold test paper of the ferrous Thiobacillus of acid oxidase is had a liking in a kind of detection according to claim 1; It is characterized in that: described nitrocellulose filter (4) is gone up also has sheep anti-mouse igg band (7); Rus monoclonal antibody II band (6) and sheep anti-mouse igg band (7) laterally arrange, and Rus monoclonal antibody II band (6) is positioned between the Rus monoclonal antibody I and sheep anti-mouse igg band (7) of colloid gold particle mark.