CN107228943B - The method for measuring bacterial content in fluid sample - Google Patents
The method for measuring bacterial content in fluid sample Download PDFInfo
- Publication number
- CN107228943B CN107228943B CN201610178405.3A CN201610178405A CN107228943B CN 107228943 B CN107228943 B CN 107228943B CN 201610178405 A CN201610178405 A CN 201610178405A CN 107228943 B CN107228943 B CN 107228943B
- Authority
- CN
- China
- Prior art keywords
- reagent tube
- filter membrane
- reagent
- fluid sample
- bacterial content
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Urology & Nephrology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Hematology (AREA)
- Medicinal Chemistry (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Food Science & Technology (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The present invention relates to a kind of methods of bacterial content in measurement fluid sample, mainly solve the problems, such as that measuring speed is slower, inconvenient, result is inaccurate in the prior art.The present invention in fluid sample in bacterial content measuring device, carries out the measurement of bacterial content in fluid sample by using a kind of method of bacterial content in measurement fluid sample;Described device includes that cabinet, Reagent Tube placement location, Reagent Tube, peep hole and Enrichment of bacteria and the protein molecule technical solution that captures equipment preferably solve the above problem, be can be used for measuring in fluid sample in bacterial content.
Description
Technical field
The present invention relates to a kind of methods of bacterial content in measurement fluid sample.
Background technique
The corrosion failure of the material as caused by the vital movement of the microorganisms such as bacterium is referred to as microbiologic(al) corrosion (MIC).MIC
It is a kind of generally existing phenomenon.All there is a large amount of bacteriums in oil gas field, oil tank, gas station, oil circuit even automotive oil tank.
Every year due to corrosion caused by economic loss at 10,000,000,000 dollars or more.Wherein most important corrosion-causing bacteria is sulfate reduction
Bacterium (SRB) causes the corrosion loss of 77% or more oil well produced in USA.SRB can be under oxygen-free environment, with having in environment
Machine object is carbon source, using the hydrogen generated in bacterial biof iotalm, is hydrogen sulfide by sulfate reduction, obtains from redox reaction
Energy, and metal is also oxidized corrosion in the process.At present there are mainly three types of the detection methods of SRB, it is respectively as follows: culture detection
Technology, microscopic techniques and biochemical Direct Inspection Technology.Culture detection technique is a kind of current widely used side
Method, principle are that sample is done gradient dilution, are cultivated on culture medium appropriate, finally according to extension rate and bacterium
Positive reaction is quantified.The main foundation of the method is the dilution-to-extinction method that American Petroleum Institute API recommends, China's oil industry
Also there is specified in more detail in standard SY/T 0532-2012 " oilfield injection water bacteria analyzing method dilution-to-extinction method ".Related patents are
Have been reported that (such as CN102329851A, CN103983636A, CN1769472A, CN200510010459), currently on the market
There are SRB special test bottle is available for purchase.The advantages of cultivating detection technique is easy to operate, low in cost, as a result accuracy
It is higher.Its major defect is that time-consuming (2-14 days), and culture strain not exclusively causes quantitative result relatively low.Once negative control
There is fungus grown in group, needs to reform experiment, it is difficult to ensure that resampling result is consistent with original sample.
Microscopic techniques combine specific stain method and fluorescence method of counting, by fluorophor selectivity terrestrial reference
Remember the surface of SRB, then counts under fluorescence microscope with quantitative.But there is still a need for Short-term Cultures to improve bacterium for such method
Class concentration, cultivation cycle was at two days or so.Although its operation is relatively easy, quantitative at low cost, the Gao Te of dye molecule is needed
The opposite sex, and the relevant information of bacterial activity cannot be provided.
Biochemical Direct Inspection Technology contains a variety of concrete modes and [closes sulfate in the sparkling oilfield sewage of refined Xia Yehaichunfan
The progress chemistry and bioengineering 2,012 29 17 of reducing bacteria detection technique], it can detecte certain of inside SRB or surface
Kind albumen is (as measured 5'-AMP sulfate reduction enzyme APS[such as with enzyme-linked immunosorbent assay ELISA method
GB2354072B, CN103983636A, EP272916A1]), it can detecte sequence of nucleic acid molecules (such as polymerase of SRB specificity
Chain reaction PCR method [CN105112543A, CN1769472A, CN200510010459], fluorescence in situ hybridization FISH method
[Wang Ming justice beam dogface Zheng Ya duckweed Zhao by Wei in green the sulfate reduction dientification of bacteria and detection method the micro- life of progress
Object magazine 2,005 25 81]), it also can detecte metabolite (such as potentiometry or indicator method measurement H of SRB2S) [Li Wan
Research-sulfide-selective electrode forensic chemistry sensor 1,991 11 of adopted Zhao from light sulfate reducing bacteria bacterium amount detection method
64].These detection method accuracys are high, specificity is high, detection cycle is short (or so a few houres) but complicated for operation, generally can only
It is tested in special biochemical test room by trained professional.Although having had already appeared utilization in the market
The kit of ELISA method rapid survey bacterial concentration, but there is still a need for high speed centrifugations etc. to operate for this method, is not suitable for live inspection
It surveys.
In conclusion lacking the on-site measurement method and equipment of a kind of quick, convenient, accurate SRB at present.This patent is retouched
What is stated is specificity, simplification, portable quantitative testing device and the side of bacterium in a kind of sample based on ELISA method
Method is not necessarily to external power supply.
Summary of the invention
That the technical problem to be solved by the present invention is to measuring speeds in the prior art is slower, inconvenient, result is inaccurate
Problem provides a kind of new method for measuring bacterial content in fluid sample.This method has the advantages that quick, convenient, accurate.
To solve the above problems, The technical solution adopted by the invention is as follows: a kind of measure bacterial content in fluid sample
Method in fluid sample in bacterial content measuring device, carries out the measurement of bacterial content in fluid sample;Described device includes
Cabinet, Reagent Tube placement location, Reagent Tube, peep hole and Enrichment of bacteria and protein molecule capture equipment, and the bacterium is rich
It includes Reagent Tube placement location, filter membrane, adsorption layer, at least two waste liquid pools, at least two that collection and protein molecule, which capture equipment,
Valve, the Reagent Tube placement location are located at top of the box, and bottom is connected to filter membrane upper space, and filter membrane lower space is divided into two
Road is connected with valve 5a arrival end all the way, and another way is connected with valve 5b arrival end, the outlet end valve 5a and adsorption layer entrance side
It is connected by pipeline, the pipeline of adsorption layer outlet side protrudes into waste liquid pool 4a, and the pipeline of the outlet end valve 5b protrudes into waste liquid pool 4b
In, the top of filter membrane upper space is equipped with metal spine, and examination can be punctured when Reagent Tube is placed on Reagent Tube placement location
The aluminium foil of agent bottom of the tube;Setting at least eight Reagent Tubes of a-h, in which: Reagent Tube a is blank pipe, to sample to be installed, Reagent Tube b
With splendid attire filter membrane flushing liquor in Reagent Tube c, Reagent Tube d is interior to contain bacterial lysate, and Reagent Tube e and interior contain of Reagent Tube f are adsorbed
Layer flushing liquor, Reagent Tube g is interior to contain the specific antibody solution for having dye molecule, contains adsorption layer flushing liquor in Reagent Tube h;
The peep hole is located at right above adsorption layer, and aperture is in top of the box;The measuring process of bacterial content includes: in fluid sample
(1) it is packed into sample in Reagent Tube a, is placed in Reagent Tube placement location, reagent bottom of the tube In Aluminium Foil Packing is by lower part
Metal tip punctures, and liquid flows into above filter membrane, and the liquid less than filter sizes enters waste liquid pool 4b after filter membrane;
(2) after the sample in Reagent Tube a all flows through filter membrane, Reagent Tube successively is changed on Reagent Tube placement location
B, Reagent Tube c repeats the operation of (1), is rinsed to filter membrane;
(3) after the completion of to the flushing of filter membrane, it is changed to Reagent Tube d on Reagent Tube placement location, repeats the operation of (1),
Enter waste liquid pool 4a after bacterial lysate in Reagent Tube d is flowed through adsorption layer;
(4) Reagent Tube e is changed on Reagent Tube placement location, Reagent Tube f is rinsed, remove bacterial lysate in
Impurity;
(5) after rinsing, it is changed to Reagent Tube g on Reagent Tube placement location, repeats the operation of (3);
(6) Reagent Tube h is changed on Reagent Tube placement location, it is extra to be rinsed by the solution in Reagent Tube h
Antibody;
(7) color that adsorption layer is observed by peep hole reads corresponding concentration compared with colorimetric card.
In above-mentioned technical proposal, it is preferable that when Reagent Tube is placed on Reagent Tube placement location, put on Reagent Tube top
Piston is set, the reagent in Reagent Tube is pressed downward in the space above filter membrane.
In above-mentioned technical proposal, it is preferable that encapsulated at the top of the Reagent Tube by filter membrane, aluminium foil is torn before use.
In above-mentioned technical proposal, it is preferable that it is described by selecting different Reagent Tubes, realize that the sample of the equipment is rich
Collection captures, flushing operation.
In above-mentioned technical proposal, it is preferable that Enrichment of bacteria and protein molecule capture integration of equipments in the cabinet.
In above-mentioned technical proposal, it is preferable that when sample is aqueous phase substance, using hydrophilic filter membrane, filter membrane size is 0.5
Micron is hereinafter, to ensure that objective microbe can be retained on filter membrane;When sample is oil phase substance, using lipophilic filter membrane.
In above-mentioned technical proposal, it is preferable that the material of adsorption layer is the material of specific adsorption protein, different retentions point
The ultrafiltration membrane or the surface-functionalized adsorbent material of APS antibody of son amount.
In above-mentioned technical proposal, it is preferable that dye molecule is specifically bound by antibody and APS, the quantity of dye molecule
It is directly proportional to APS;Dye molecule is gold nano grain, other particles or the coloured molecule of other bands.
In the present invention, Enrichment of bacteria is to flow through filter membrane by sample to realize.Aluminium foil above Reagent Tube is torn, empty examination is taken
Agent pipe a, is put into 1mL sample to be tested, and after a is put into Reagent Tube placement location 1, sample bottom of the tube In Aluminium Foil Packing is by lower metal point
It punctures, liquid flows into 2 top of filter membrane, applies pressure above at this time, pushes liquid through filter membrane, greater than the solid of filter sizes
Substance is left on filter membrane, liquid outflow.Valve 5b is opened at this time, and valve 5a is closed, and waste liquid flows out to right side waste liquid pool 4b.When
When sample is the aqueous phase substances such as oil field waste, using hydrophilic filter membrane, filter membrane size should be at 0.5 micron or less to ensure that target is micro-
Biology can be retained on filter membrane.When sample is the oil phase substances such as product oil, using lipophilic filter membrane.When sample all flows through
After filter membrane, sample cell is pulled out, takes out upper piston, the Reagent Tube b for being preset in front-seat is put into position 1, sample bottom of the tube aluminium foil
Packaging is broken by lower metal spine, is put into piston, repeats above-mentioned pressurization process to rinse filter membrane.Continue substitute be Reagent Tube c into
Row rinses.After flushing, Reagent Tube d is added, includes bacterial lysate, waiting starts to pressurize after a certain period of time, at this time valve
Door 5b is closed, and valve 5a is opened, and bacterial lysate enters left side waste liquid pool 4a after flowing through adsorption layer 3.The material of adsorption layer 3 can be with
It is selected as the material of specific adsorption protein, the ultrafiltration membrane of different molecular weight cut off, the surface-functionalized absorption of APS antibody
Material etc..Continue to replace Reagent Tube e, Reagent Tube f and be rinsed, this step purpose is to remove to remove (target) egg in bacterial lysate
Impurity outside white matter.G is the specific antibody solution with dye molecule, and dye molecule is specifically bound by antibody and APS,
The quantity of dye molecule is directly proportional to APS.Dye molecule can be gold nano grain, other particles, coloured point of other bands
Son etc..The extra antibody not in conjunction with APS is finally washed out by the solution in Reagent Tube h.The position and top of adsorption layer 3 are observed
Hole i is corresponding, in the corresponding colorimetric card of the 3 preset various concentration in position side of adsorption layer as quantitative reference.
The present apparatus contains Enrichment of bacteria, protein molecule captures and reads three units, and being that one kind is integrated carries
Formula equipment, can be realized in-site measurement.Enrichment of bacteria unit realized in such a way that sample flow is through filter membrane bacterium separation and
Enrichment, required time greatly shorten compared with cultivation.The capture of protein molecule is to break cell wall release by lysate
Out after APS, APS is enriched on adsorption layer, dye granule is further then enriched with by antibody-antigene specific effect, is contaminated
The concentration of material and the concentration of APS are directly proportional, finally by the concentration of the color instruction SRB of adsorption layer, achieve preferable technology effect
Fruit.
Detailed description of the invention
Fig. 1 is the appearance diagram of device of the present invention;
Fig. 2 is each modular structure schematic diagram inside device of the present invention;
In Fig. 1, Fig. 2,1 is Reagent Tube placement location;2 be filter membrane;3 be adsorption layer;4a, 4b are waste liquid pool;5a, 5b are valve
Door;A is Reagent Tube, blank pipe, sample to be installed;B, c is Reagent Tube, contains filter membrane flushing liquor;D is Reagent Tube, contains bacteria lysis
Liquid;E, f is Reagent Tube, contains adsorption layer flushing liquor;G is Reagent Tube, contains the specific antibody solution for having dye molecule;h
For Reagent Tube, adsorption layer flushing liquor is contained;I is peep hole.
The present invention will be further described below by way of examples, but is not limited only to the present embodiment.
Specific embodiment
[embodiment 1]
A kind of method of bacterial content in measurement fluid sample, as shown in Figure 1 and Figure 2.Bacterial content is surveyed in fluid sample
It measures on device, carries out the measurement of bacterial content in fluid sample;Described device include cabinet, Reagent Tube placement location, Reagent Tube,
Peep hole and Enrichment of bacteria and protein molecule capture equipment, and it includes examination that the Enrichment of bacteria and protein molecule, which capture equipment,
Agent pipe placement location, filter membrane, adsorption layer, at least two waste liquid pools, at least two valves, the Reagent Tube placement location are located at case
At the top of body, bottom is connected to filter membrane upper space, and filter membrane lower space is divided into two-way, is connected all the way with valve 5a arrival end, separately
It is connected all the way with valve 5b arrival end, the outlet end valve 5a is connected with adsorption layer entrance side by pipeline, adsorption layer outlet side
Pipeline protrudes into waste liquid pool 4a, and the pipeline of the outlet end valve 5b protrudes into waste liquid pool 4b, and the top of filter membrane upper space is equipped with gold
Belong to spine, the aluminium foil of reagent bottom of the tube can be punctured when Reagent Tube is placed on Reagent Tube placement location;A-h eight examinations of setting
Agent pipe, in which: Reagent Tube a is blank pipe, to sample to be installed, contains filter membrane flushing liquor, Reagent Tube d in Reagent Tube b and Reagent Tube c
Adsorption layer flushing liquor is contained in interior splendid attire bacterial lysate, Reagent Tube e and Reagent Tube f, is contained in Reagent Tube g and is had dye molecule
Specific antibody solution, adsorption layer flushing liquor is contained in Reagent Tube h;The peep hole is located at right above adsorption layer, and aperture exists
Top of the box;The peep hole is located at right above adsorption layer, and aperture is in top of the box.
Device of the present invention is integrated in the ganoine packing box of 0.3m*0.2m*0.2m, and user only needs according to explanation
Book carries out injection operation.
Front-seat Reagent Tube (a-h) is preset cracking, washing, label solution, packaged with aluminium foil when being not used, when use
Tear aluminium foil, be placed in Reagent Tube placement location 1 one by one to specifications so that different solutions sequentially flow through filter membrane and
Adsorption layer.The position i is peep hole, for observing the color of adsorption layer, and compared with preset reference colorimetric card, to read sample
The concentration of SRB in product.
In the present invention, Enrichment of bacteria is to flow through filter membrane by sample to realize.Aluminium foil above Reagent Tube is torn, empty examination is taken
Agent pipe a, is put into 1mL sample to be tested, and after a is put into designated position Reagent Tube placement location 1, sample bottom of the tube In Aluminium Foil Packing is by under
Portion's metal tip punctures, and liquid flows into 2 top of filter membrane, applies pressure above at this time, pushes liquid through filter membrane, is greater than filter hole
The solid matter of diameter is left on filter membrane, liquid outflow.Valve 5b is opened at this time, and valve 5a is closed, and it is useless that waste liquid flows out to right side
Liquid pool 4b.When sample is the aqueous phase substances such as oil field waste, using hydrophilic filter membrane, filter membrane size should 0.5 micron or less with
Ensure that objective microbe can be retained on filter membrane.When sample is the oil phase substances such as product oil, using lipophilic filter membrane.Work as sample
After product all flow through filter membrane, sample cell is pulled out, takes out upper piston, the Reagent Tube b for being preset in front-seat is put into position 1, sample
Bottom of the tube In Aluminium Foil Packing is broken by lower metal spine, is put into piston, repeats above-mentioned pressurization process to rinse filter membrane.Continue to substitute and is
Reagent Tube c is rinsed.After flushing, Reagent Tube d is added, includes bacterial lysate, waiting starts to add after a certain period of time
Pressure, valve 5b is closed at this time, and valve 5a is opened, and bacterial lysate enters left side waste liquid pool 4a after flowing through adsorption layer 3.Adsorption layer 3
Material can choose the material for specific adsorption protein, the ultrafiltration membrane of different molecular weight cut off, surface-functionalized APS
The adsorbent material etc. of antibody.Continue to replace Reagent Tube e, Reagent Tube f and be rinsed, this step purpose is to remove in bacterial lysate
Impurity in addition to (target) protein.G is the specific antibody solution with dye molecule, and dye molecule passes through antibody and APS
Specific binding, the quantity of dye molecule are directly proportional to APS.Dye molecule can be gold nano grain, other particles, other bands
Coloured molecule etc..The extra antibody not in conjunction with APS is finally washed out by the solution in Reagent Tube h.The position of adsorption layer 3
It is corresponding with top peep hole i, in the corresponding colorimetric card of the 3 preset various concentration in position side of adsorption layer as quantitative reference.
In order to keep advantages of the present invention, technical solution clearer, clear, combined with specific embodiments below to of the invention
Step elaborates.
The quantitative detection of SRB in waste water
1) Open valve 5b closes valve 5a;
2) Reagent Tube is taken, top aluminium foil encapsulation is torn, removes sky Reagent Tube a, be packed into 1mL sample, put it into Reagent Tube
Placement location 1, is put into piston, pushes down on piston and flows through filter membrane;
3) after whole liquid flow through filter membrane, Reagent Tube a is removed, piston is removed, Reagent Tube b is put into Reagent Tube and places position
1 is set, pushes piston that liquid in Reagent Tube b is made all to flow through filter membrane;
4) 2) operation in is repeated to Reagent Tube c;
5) Open valve 5a closes valve 5b;
6) Reagent Tube d is taken, is slightly rocked after putting it into Reagent Tube placement location 1, the position for contacting bottom aluminium foil with needle
There is gap, make liquid flow above filter membrane, stands 10 minutes;Push piston, coutroi velocity make liquid slowly flow through filter membrane and
Adsorption layer;
7) after whole liquid flow through filter membrane, Reagent Tube d is removed, piston is removed, Reagent Tube e is put into Reagent Tube and places position
1 is set, pushes piston that liquid in Reagent Tube e is made all to flow through filter membrane;
8) 7) operation in is repeated to Reagent Tube f;
9) Reagent Tube g is taken, 1 pusher piston of Reagent Tube placement location is put it into, coutroi velocity makes liquid rest on suction
Before attached layer 3,10 minutes are stood;Piston is pushed, coutroi velocity makes liquid slowly flow through adsorption layer;
10) after whole liquid flow through filter membrane, Reagent Tube g is removed, piston is removed, Reagent Tube h is put into Reagent Tube and is placed
Position 1 pushes piston that liquid in Reagent Tube h is made all to flow through filter membrane;
11) color for observing adsorption layer 3 reads corresponding concentration compared with colorimetric card.
In the present embodiment, being put into sample is that 1mL contains 107The aqueous solution of CFU/mL SRB, operation total duration are 1 hour
30 minutes or so, compared with colorimetric card, finally being measured concentration was 5 × 106CFU/mL-107Between CFU/mL, with sample value phase
Together.
It should be noted that those skilled in the art can also make such or such appearance under the introduction of this specification
Easy variation pattern, such as equivalent way or obvious mode of texturing, should all be within protection scope of the present invention.
Claims (7)
1. the method for bacterial content, in fluid sample in bacterial content measuring device, carries out liquid in a kind of measurement fluid sample
The measurement of bacterial content in body sample;Described device includes cabinet, Reagent Tube placement location, Reagent Tube, peep hole and bacterium
Enrichment and protein molecule capture equipment, and it includes filter membrane, adsorption layer, at least that the Enrichment of bacteria and protein molecule, which capture equipment,
Two waste liquid pools, at least two valves, the Reagent Tube placement location are located at top of the box, and bottom and filter membrane upper space connect
Logical, filter membrane lower space is divided into two-way, is connected all the way with valve (5a) arrival end, and another way is connected with valve (5b) arrival end,
The valve outlet end (5a) is connected with adsorption layer entrance side by pipeline, and the pipeline of adsorption layer outlet side protrudes into waste liquid pool (4a),
The pipeline of the outlet end valve (5b) protrudes into waste liquid pool (4b), and the top of filter membrane upper space is equipped with metal spine, works as Reagent Tube
The aluminium foil of reagent bottom of the tube can be punctured when being placed on Reagent Tube placement location;At least eight Reagent Tubes are set, in which: reagent
Managing (a) is blank pipe, to sample to be installed, filter membrane flushing liquor is contained in Reagent Tube (b) and Reagent Tube (c), is contained in Reagent Tube (d)
Adsorption layer flushing liquor is contained in bacterial lysate, Reagent Tube (e) and Reagent Tube (f), is contained in Reagent Tube (g) and is had dye molecule
Specific antibody solution, adsorption layer flushing liquor is contained in Reagent Tube (h);The peep hole is located at right above adsorption layer, aperture
In top of the box;When Reagent Tube is placed on Reagent Tube placement location, piston is placed on Reagent Tube top, it will be in Reagent Tube
Reagent presses downward in the space above filter membrane;The measuring process of bacterial content includes: in fluid sample
(1) it is packed into sample in Reagent Tube (a), is placed in Reagent Tube placement location, reagent bottom of the tube In Aluminium Foil Packing is by lower part gold
It is broken to belong to spine, liquid flows into above filter membrane, and the liquid less than filter sizes enters waste liquid pool (4b) after filter membrane;
(2) after the sample in Reagent Tube (a) all flows through filter membrane, Reagent Tube successively is changed on Reagent Tube placement location
(b), Reagent Tube (c) repeats the operation of (1), is rinsed to filter membrane;
(3) after the completion of to the flushing of filter membrane, it is changed to Reagent Tube (d) on Reagent Tube placement location, repeats the operation of (1), it will
Bacterial lysate in Reagent Tube (d) enters waste liquid pool (4a) after flowing through adsorption layer;
(4) Reagent Tube (e) is changed on Reagent Tube placement location, Reagent Tube (f) is rinsed, remove bacterial lysate in
Impurity;
(5) after rinsing, it is changed to Reagent Tube (g) on Reagent Tube placement location, repeats the operation of (3);
(6) it is changed to Reagent Tube (h) on Reagent Tube placement location, it is extra to be rinsed by the solution in Reagent Tube (h)
Antibody;
(7) color that adsorption layer is observed by peep hole reads corresponding concentration compared with colorimetric card.
2. measuring the method for bacterial content in fluid sample according to claim 1, it is characterised in that at the top of the Reagent Tube
It is encapsulated by aluminium foil, the aluminium foil above Reagent Tube is torn before use.
3. measuring the method for bacterial content in fluid sample according to claim 1, it is characterised in that described by selecting not
Same Reagent Tube, realizes example enrichment, capture, the flushing operation of the equipment.
4. measuring the method for bacterial content in fluid sample according to claim 1, it is characterised in that Enrichment of bacteria and albumen
Matter molecule captures integration of equipments in the cabinet.
5. measuring the method for bacterial content in fluid sample according to claim 1, it is characterised in that when sample is aqueous-phase material
When matter, using hydrophilic filter membrane, filter membrane size is at 0.5 micron hereinafter, to ensure that objective microbe can be retained on filter membrane;When
When sample is oil phase substance, using lipophilic filter membrane.
6. measuring the method for bacterial content in fluid sample according to claim 1, it is characterised in that the material of adsorption layer is
The ultrafiltration membrane of the material of specific adsorption protein, different molecular weight cut off.
7. measuring the method for bacterial content in fluid sample according to claim 1, it is characterised in that dye molecule passes through anti-
Body and 5'-AMP sulfate reduction enzyme APS are specifically bound, and the quantity of dye molecule is directly proportional to APS.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610178405.3A CN107228943B (en) | 2016-03-25 | 2016-03-25 | The method for measuring bacterial content in fluid sample |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610178405.3A CN107228943B (en) | 2016-03-25 | 2016-03-25 | The method for measuring bacterial content in fluid sample |
Publications (2)
Publication Number | Publication Date |
---|---|
CN107228943A CN107228943A (en) | 2017-10-03 |
CN107228943B true CN107228943B (en) | 2018-12-04 |
Family
ID=59932652
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201610178405.3A Active CN107228943B (en) | 2016-03-25 | 2016-03-25 | The method for measuring bacterial content in fluid sample |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN107228943B (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110174382B (en) * | 2019-05-23 | 2020-11-20 | 安徽维嵩生产力促进有限公司 | Device for detecting sewage bacteria concentration |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0272916A1 (en) * | 1986-12-23 | 1988-06-29 | Conoco Phillips Company | Sulfate-reducing bacteria determination and control |
WO2014042933A1 (en) * | 2012-09-14 | 2014-03-20 | Halliburton Energy Services, Inc. | Systems and methods for analyzing microbiological substances |
CN103983636A (en) * | 2014-05-20 | 2014-08-13 | 中国石油化工股份有限公司 | Rapid detecting method of sulphate reducing bacteria and kit thereof |
CN104312962A (en) * | 2014-10-27 | 2015-01-28 | 中国石油化工股份有限公司 | Separation and purification method for sulfate reducing bacteria in sewage of oil field |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20040091882A1 (en) * | 1999-03-24 | 2004-05-13 | Michel Magot | Method of detecting sulphate-reducing bacteria |
-
2016
- 2016-03-25 CN CN201610178405.3A patent/CN107228943B/en active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0272916A1 (en) * | 1986-12-23 | 1988-06-29 | Conoco Phillips Company | Sulfate-reducing bacteria determination and control |
WO2014042933A1 (en) * | 2012-09-14 | 2014-03-20 | Halliburton Energy Services, Inc. | Systems and methods for analyzing microbiological substances |
CN103983636A (en) * | 2014-05-20 | 2014-08-13 | 中国石油化工股份有限公司 | Rapid detecting method of sulphate reducing bacteria and kit thereof |
CN104312962A (en) * | 2014-10-27 | 2015-01-28 | 中国石油化工股份有限公司 | Separation and purification method for sulfate reducing bacteria in sewage of oil field |
Also Published As
Publication number | Publication date |
---|---|
CN107228943A (en) | 2017-10-03 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN102243234B (en) | Automatic bacterial sorting and labeling method based on immune method, and apparatus thereof | |
KR20180048558A (en) | Components, devices and methods of devices for purifying and testing biomolecules from biological samples | |
CN109444070A (en) | Oil content all automatic measurement instrument in water | |
JP2012513773A5 (en) | ||
EP2831577B1 (en) | Methods and systems useful for foodborne pathogen detection | |
JP2004226403A (en) | Protein detecting flow system and protein detecting method | |
DE602005014835D1 (en) | Method, apparatus, reagent kit and reagent for distinguishing erythrocytes in biological samples | |
CN106238112A (en) | A kind of micro-fluidic chip and the application in the qualification and drug sensitive experiment of pathogen thereof | |
CN205115443U (en) | Collection draws, amplifys and detects integrated gene detection device | |
JP2018522202A (en) | Cartridge for dispensing particles and reagent fluids | |
CN202049159U (en) | Automatic bacterium sorting and labeling device based on immune method | |
CN102634448B (en) | Biological sampling swab | |
CN202558865U (en) | Biology sampling swab | |
CN205730480U (en) | A kind of two-part plankton settler | |
CN107228943B (en) | The method for measuring bacterial content in fluid sample | |
CN111289762A (en) | Micro-fluidic chip sample adding device and testing method | |
CN105018343B (en) | Ortho states microorganism oil gas and the automation sample processing device and Automation workstation of hydrate exploration technology | |
JP5750796B2 (en) | Diatom detection method | |
WO2009123772A3 (en) | Device and method for automating microbiology processes | |
CN110023758A (en) | For separating equipment, system, method and the external member of analyte from body fluid sample | |
ES2656117T3 (en) | Method and apparatus for the detection of microorganisms | |
CN202530081U (en) | Bacterium sampling swab | |
CN107219361B (en) | Measure the device of bacterial content in fluid sample | |
CN211826105U (en) | Micro-fluidic chip application of sample device | |
CN107216998B (en) | Automatic device for rapidly measuring bacterial content in oil field sewage and finished oil |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |