CN107402299A - One kind centrifuges detection means - Google Patents
One kind centrifuges detection means Download PDFInfo
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- CN107402299A CN107402299A CN201610330378.7A CN201610330378A CN107402299A CN 107402299 A CN107402299 A CN 107402299A CN 201610330378 A CN201610330378 A CN 201610330378A CN 107402299 A CN107402299 A CN 107402299A
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- Prior art keywords
- immobilon
- detection means
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- centrifugal
- centrifugation
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- 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
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
Abstract
The invention discloses one kind to centrifuge detection means.It includes sample introduction part, immobilon-p, centrifugal device and detector；The centrifugal device is included by the centrifugal rotor and support base of motor driving, and the centrifugal rotor is using the support base as support；The sample introduction part is located at the middle part of the centrifugal rotor, and is connected with centrifugal rotor；The sample introduction part includes liquid phase storage device and sample feeding pipe；The liquid phase storage device is connected with the sample feeding pipe；The immobilon-p is connected on the centrifugal rotor with the sample feeding pipe；The detector is located at the one or both sides of the immobilon-p.The present invention centrifuge detection means have detection speed fast, high sensitivity, it is easy to use the characteristics of.
Description
Technical field
The present invention relates to one kind to centrifuge detection means, belongs to Instrumental Analysis field.
Background technology
Immunology detection technology is measure antigen, antibody, immunocyte and the chemical composition of applied immunology principle design
Deng laboratory facilities, be widely used in from human body and animal body can carry out medical diagnosis on disease and health detection sample and
For environment, Pharmaceutical Analysis, food and the sample of Industrial Analysis.Conventional has immune turbidity technology, immobilized enzyme to be immunized
Determination techniques, chemiluminescence detection technology, immunofluorescence label technology, flow cytometry, colloidal gold technique etc..
Immune turbidity technology, also referred to as immune turbidimetry is soluble antigen, antibody specific bond in the liquid phase, produces one
Determine the compound of size, form the refraction or absorption of light, determine the transmitted light after this refraction or absorption or scattering light is made
For unit of account, for quantitatively detecting, but detection sensitivity is low, is not suitable for trace detection.Solid-phase enzyme immunoassay
Immobilization and antigen or antibody of the technology based on antigen or antibody enzyme mark, be incorporated in surface of solid phase carriers antigen or
Antibody keeps its immunologic competence, and the enzyme conjugates of antigen or antibody had both retained its immunologic competence, retains the work of enzyme again
Property, in measure, by examining sample (determining antibody or antigen therein) and enzyme-labelled antigen or antibody by different steps
Suddenly the antigen or antibody with surface of solid phase carriers react, and have high sensitivity, linear response range is wide and is easily achieved
The remarkable advantages such as automation, but detect reaction time length and limit its use.Immunochemiluminescence detection technique is a kind of
Highly sensitive micro and Analytical Methods of Trace, have easy to operate, high sensitivity, linear response range wide and be easy to real
The now remarkable advantage such as automation, is widely used in environment, clinic, Pharmaceutical Analysis, food and Industrial Analysis,
It is solid phase separation means and antigen based on antigen or antibody or the luminescence reagent labelling technique of antibody, but when detecting reaction
Between long and on detection device requirement height also influence its use.Immunofluorescence label technology, flow cytometry, collaurum
Technology is also that conventional detection technique is widely used, but has its corresponding deficiency, and the detection reaction time is long or sensitive
Degree, accuracy shortcoming are the deficiencies of generally existing.
High sensitivity, quick, miniaturization, quantitative, automation is the development of current clinical immunization detection technique product entirely
Trend, but existing above-mentioned function can not be all realized simultaneously.Therefore, exploitation one kind can realize both have high sensitivity,
Complete quantitative, automation feature, while have that detection is quick, the simple new detection device of equipment again, not only user
Just, reduce and waste, while be also remarkably improved operating efficiency, there is weight in the numerous areas for detecting and analyzing, separate
The Practical significance wanted.
The content of the invention
It is an object of the invention to provide one kind to centrifuge detection means, and the present invention, which centrifuges detection means, has detection
Speed is fast, high sensitivity, it is easy to use the characteristics of.
Centrifugation detection means provided by the invention, it includes sample introduction part, immobilon-p, centrifugal device and detector；
The centrifugal device is included by the centrifugal rotor and support base of motor driving, and the centrifugal rotor is with described
Support base is support；
The sample introduction part is located at the middle part of the centrifugal rotor, and is connected with centrifugal rotor；The sample introduction part bag
Include liquid phase storage device and sample feeding pipe；The liquid phase storage device is connected with the sample feeding pipe；
The immobilon-p is connected on the centrifugal rotor with the sample feeding pipe；
The detector is located at the one or both sides of the immobilon-p.
In above-mentioned centrifugation detection means, the immobilon-p is placed in immobilon-p apparatus for placing, and the immobilon-p
Apparatus for placing is on the centrifugal rotor；
The immobilon-p and the centrifugal rotor are dismountable structure.
In above-mentioned centrifugation detection means, the immobilon-p apparatus for placing is rotary moving fixing device and/or groove
Shape squash type fixing device
In above-mentioned centrifugation detection means, the immobilon-p is fixed by immobilon-p fixator, and the immobilon-p is consolidated
Determine device and be selected from immobilon-p support egative film sample part, lateral flow test strips buckle sample part and hyalomitome embedding formula part up and down
At least one of.
In above-mentioned centrifugation detection means, the hyalomitome embeds formula part as up and down the two of the immobilon-p up and down
Side is covered by hard transparent material, and the area of the hard transparent material of the immobilon-p respective side is more than or equal to institute
State the area of immobilon-p.
In above-mentioned centrifugation detection means, the immobilon-p is additionally provided with therewith in the one end being connected with the sample feeding pipe
The liquid adsorption discrete part being connected, the immobilon-p are logical provided with being attached thereto in the distal end of the centrifugal rotor
Liquid collecting member, the liquid adsorption discrete part are connected with the sample feeding pipe.
In above-mentioned centrifugation detection means, the sample introduction part includes sampling pump, described in the sampling pump driving
Liquid in liquid phase storage device enters the sample feeding pipe.
In above-mentioned centrifugation detection means, the liquid phase storage device includes measuring samples storage device and detection phase
Storage device；
The measuring samples storage device and the detection phase storage device are connected with the sample feeding pipe, and by institute
State sampling pump driving.
In above-mentioned centrifugation detection means, the liquid phase storage device also includes cleaning fluid storage device, described clear
Washing lotion storage device is connected with the sample feeding pipe, and is driven by the sampling pump.
In above-mentioned centrifugation detection means, the centrifugal rotor is using plane or by the outward-dipping type in center；
The centrifugal device is provided with a shell.
In above-mentioned centrifugation detection means, the rotary moving fixing device is to be set on the centrifugal rotor
Columnar projections attachment means, the immobilon-p fixator is provided with to match with the attachment means of the columnar projections
Poroid part.
In above-mentioned centrifugation detection means, the centrifugal device, the sample introduction part and the detector are equipped with
Presetting apparatus.
In above-mentioned centrifugation detection means, the sample feeding pipe is connected with the immobilon-p using dismountable；
The liquid adsorption discrete part is connected with the sample feeding pipe using dismountable.
In above-mentioned centrifugation detection means, the material of the immobilon-p is using nitrocellulose filter, Kynoar
Any of film, nylon membrane and DEAE cellulose membranes, and the one or both sides of the immobilon-p are provided with backing；
The liquid adsorption discrete part includes colloid gold label adsorbed film, fluorescent labeled antibody adsorbed film, chemiluminescence
Mark at least one of adsorbed film, poly ester fiber dispersion membrane and glass fibre dispersion membrane；
The detector includes any of detector of absorbance, fluorescence, chemiluminescence and image processing.
In the present invention, the fluid collection device can be made of water-absorbing material, and specially blotting paper and/or water suction is solidifying
Glue, the fluid collection device can be liquid collecting container.
In above-mentioned centrifugation detection means, the plane interlude of the centrifugal rotor is provided with hole and/or provided with hyalomere
Part, and the hole and/or the transparent component make the immobilon-p be directly exposed to the detector.
The application of the present invention that detection means is centrifuged in immunoassay product detection, specifically, the immune production
Product include at least one of antigen, antibody, immunocyte and chemical composition.
The present invention has advantages below：
1st, the present invention drives detection liquid phase to be flowed on immobilon-p and cleaning step with centrifugal device, improves thing to be checked
Capture binding ability, reduce immobilon-p ambient noise interference, improve detection sensitivity, realize it is existing detection examination
The high-sensitivity detection of agent.2nd, it is of the invention to drive detection liquid phase to be flowed on immobilon-p with centrifugal device, change existing
The present situation that some film detection techniques flow by nature and liquid reduces with the extension of flow on film, can keep
Liquid at the uniform velocity flows on film, ensure that the homogeneity that thing to be checked combines on film, can improve detection accuracy.3rd, originally
Invention drives detection liquid phase to be flowed on immobilon-p with centrifugal device, maintains liquid and is at the uniform velocity flowed on film, is shortened
Detection time is quick.Overcome existing film detection technique by nature flowing, flowing velocity of the liquid on film with
Time and the problem of of slowing down, complete the deficiency that a detection generally requires the time of more than 15 minutes.4th, it is of the invention
With centrifugal device driving liquid flowing and sampling pump sample introduction, operating procedure is simple, is easy to exploitation more easily to minimize
Detection device.Overcome in existing high-sensitivity detection technology using multi-step, too many levels drive control, be related to
Detect the deficiency of transposition and the movement of sample, detection phase and reaction carriers.5th, it is of the invention that liquid is driven with centrifugal device
Body flows and sampling pump sample introduction, and the integrated design of detector and centrifugal device, and operating procedure is simple, equipment knot
Structure is simple, it is easy to accomplish automation mechanized operation.Therefore, the present invention has high sensitivity, full the characteristics of quantifying, automating,
Simultaneously again with detection it is quick, use the simple detection technique of equipment；Waste that is not only easy to use, reducing raw material,
Operating efficiency is also significantly improved simultaneously, applied to detection and analysis, the numerous areas of separation.
Brief description of the drawings
Fig. 1 is the schematic diagram that detection means is centrifuged in the embodiment of the present invention 1.
Fig. 2 is the schematic diagram that Fig. 1 increases part.
Fig. 3 is the schematic diagram that hole and/or transparent component centrifugation structure of the detecting device are provided with the embodiment of the present invention 2.
Fig. 4 is that immobilon-p supports egative film sample modular construction schematic diagram.
Fig. 5 is the fixer structure schematic diagram of immobilon-p in Fig. 4.
Fig. 6 is immobilon-p fixator rotary moving attachment means structural representation in Fig. 5.
Fig. 7 is the schematic diagram that centrifugal rotor uses outward-dipping type structure in Fig. 1.
Fig. 8 is immobilon-p and sample feeding pipe attachment structure schematic diagram.
Fig. 9 is centrifugal rotor immobilon-p rotary moving attachment means structural representation.
Figure 10 is the structural representation that immobilon-p fixator hyalomitome embeds formula part up and down.
Marked in figure as follows：
1 sample introduction part；2 immobilon-ps；3 centrifugal devices；4 detectors；5 shells；6 liquid phase storage devices；7
Sampling pump；8 sample feeding pipes；9 centrifugal rotors；The outward-dipping type centrifugal rotor in 9A centers；10 motors；11 supports
Base；12 holes and/or transparent component；Detector on 13；14 times detectors；15 liquid adsorption discrete parts；16
Liquid collecting member；17 immobilon-ps support egative film；18 poroid parts；19 point sample grooves；20 lateral flow test strips buckles
Sample part；21 observation windows；22 liquid collecting members export；23 columnar projections；24 sample introduction tube fixers；25 liquid
Phase；26 immobilon-p apparatus for placing；27 hard transparent lower cover slips；28 hard transparent upper cover plates；Naked empty interlayer before 29；
Naked empty interlayer after 30.
Embodiment
The present invention is further described below in conjunction with the accompanying drawings, but the invention is not limited in following examples.
The preparation of embodiment 1, horizontal type centrifugal separating device
Include as shown in figure 1, the present invention centrifuges detection means：Sample introduction part 1, immobilon-p 2, centrifugal device 3
With detector 4.
As shown in Fig. 2 centrifugal device 3 includes centrifugal rotor 9, motor 10 and the support for supporting centrifugal rotor 9
Base 11, motor 10 drive centrifugal rotor 9 to rotate.In order to protect centrifugal device, it can be located at a shell 5
In.
As shown in Fig. 2 sample introduction part 1 is included by liquid phase storage device 6, sampling pump 7 and sample feeding pipe 8.Liquid phase stores
Device 6 is connected with sample feeding pipe 8, and sample feeding pipe 8 is driven into located at the middle part of centrifugal rotor 9, and by sampling pump 7
In；Sample feeding pipe 8 is on centrifugal rotor 9.In order to control the sample introduction speed of the speed of centrifugal device and sampling device,
Centrifugal device and sample introduction part are connected with the part with programme-control speed.
As shown in Figure 2 and Figure 4, immobilon-p 2 and centrifugal rotor 9 are dismountable structure；Immobilon-p 2, which is positioned over, to be set
In the immobilon-p apparatus for placing 26 of the outer of centrifugal rotor 9, immobilon-p 2 in centrifugal rotor 9 proximal part be provided with
The liquid adsorption discrete part 15 being connected, immobilon-p 2 in centrifugal rotor distal end be provided be attached thereto logical liquid
Body collecting part 16, liquid adsorption discrete part 15 are connected with sample feeding pipe 8.Wherein, the material of immobilon-p 2 is adopted
With any of nitrocellulose filter, polyvinylidene fluoride film, nylon membrane and DEAE cellulose membranes, and immobilon-p 2
One or both sides be provided with backing；Liquid adsorption discrete part 15 includes colloid gold label adsorbed film, fluorescent labeled antibody
At least one of adsorbed film, chemiluminescent labeling adsorbed film and dispersion membrane；Fluid collection device 16 uses water imbibition material
Material, such as blotting paper and/or water absorbent gel, it is also possible to liquid collecting container.
As shown in Figure 4, Figure 5, a kind of immobilon-p fixing device set for fixed immobilon-p 2, as supported in Fig. 4
PVC board, transparent plastic sheet, poly (methyl methacrylate) plate etc. may be selected in egative film 17, support egative film 17.Immobilon-p is consolidated in Fig. 5
It is lateral flow test strips buckle sample part 20 to determine device, specifically includes poroid part 18, point sample groove 19, observation window 21
With liquid collecting member outlet 22.After solid phase membrane structure is put into lateral flow test strips buckle spline structure 20, point sample groove
19 corresponding positions are liquid adsorption discrete part, and the corresponding position of observation window 21 is immobilon-p 2, and liquid collecting member goes out
22 corresponding positions of mouth are liquid collecting member 16 (for water-absorbing material or liquid collecting container).
As shown in Fig. 6, Fig. 9, immobilon-p apparatus for placing 26 uses rotary moving fixing device, and it is convex that it includes column
23 are played, columnar projections 23 match with the poroid part 18 set on lateral flow test strips buckle sample part 20.Post
Shape projection 23 is the column structure being arranged on centrifugal rotor 9.By the columnar projections on centrifugal rotor 9 during use
In the poroid part 18 of 23 insertion lateral flow test strips buckle sample parts 20, one end of sample feeding pipe 8 is connected to point sample
Groove 19, the other end are connected with stored in liquid phase device 6, and liquid phase is driven into sample feeding pipe 8 through sampling pump, then adds to point sample
Groove 19.It is evenly distributed with centrifugal rotor 9 for fixing immobilon-p fixator (such as lateral flow by poroid part 18
Test strips buckle sample part 20) columnar projections 23.
As shown in Figure 1 and Figure 2, detector 4 is located at the outer of centrifugal rotor 9, and it is located at the side of immobilon-p 2.
As shown in figure 8, immobilon-p 2 and the attachment structure of sample feeding pipe 8, sample feeding pipe 8 is placed in the top of immobilon-p 2,
Connection is directly contacted with liquid adsorption discrete part 15 or contact connection is added dropwise by liquid phase 25.Liquid phase 25 passes through sample introduction
Pipe 8 is loaded directly into or is added drop-wise to liquid adsorption discrete part 15.
As shown in Figure 10, another immobilon-p fixing device set for fixed immobilon-p 2, specifically includes hard transparent
Lower cover slip 27, hard transparent upper cover plate 28, preceding naked empty interlayer 29 and rear naked empty interlayer 30.During detection liquid phase by from
The heart, premenstrual naked empty interlayer 29, into liquid adsorption discrete part 15, flow through immobilon-p 2, reacted liquid phase is from rear
Naked empty interlayer 30 is discharged.
The preparation of embodiment 2, the outward-dipping type centrifugal separating device in center
As shown in fig. 7, the outward-dipping type in center of the present invention centrifuges detection means and horizontal type centrifugal separating device institute
It is identical containing variety of components, including sample introduction part 1, immobilon-p 2, centrifugal device 3 and detector 4 and corresponding composition.
Difference is that centrifugal rotor 9 is led to using outward-dipping type centered on the outward-dipping type centrifugal rotor 9A in center, immobilon-p
Lateral flow test strips buckle sample part 20 is crossed to place in outwardly sloping type on the outward-dipping type centrifugal rotor 9A in center,
It can be centrally disposed in outward-dipping type centrifugal rotor 9A outer surface or the internal centrifugal hole or interlayer set.
Embodiment 3, the present invention are provided with the preparation of hole and/or transparent component centrifugal separating device
As shown in figure 3, it is identical with being set in embodiment 1, unlike：By the plane interlude of centrifugal rotor 9
Provided with hole and/or transparent component 12, and hole and/or transparent component 12 make upper detector of the immobilon-p 2 directly with detector
13rd, lower detector 14 is relative, i.e., hole and/or transparent component 12 make immobilon-p 2 exposed to detector upper detector 13,
Between lower detector 14, so that upper detector 13, lower detector 14 can read the detection data of immobilon-p 2 simultaneously.
The application experiment of the present invention：
The Detection results of experiment one, the preparation of the planar-type centrifugal separating device of the embodiment of the present invention 1 and device：
First, the preparation of device
1st, experiment material
Planetary reducing motor (i.e. the motor of centrifugal device, 500-5000 revs/min of output speed, power 60w, customization),
Potentiometer, thickness 1.5mm iron plates, miniature peristaltic pump (sharp pump industry, model BW100 are created in the sampling pump i.e. in sample introduction part, Baoding),
Myoglobins colloidal-gold detecting-card (the i.e. integrative-structure of the immobilon-p of immobilon-p and immobilon-p fixator, the production of Changzhou Bo Wendi companies
Product), human muscle hemoglobin (i.e. measuring samples, Sigma-Aldrich products, catalog number F3879-1G), collaurum are quantitative
Chromatographic analysis instrument (i.e. detector, Norway's Skannex products).
2nd, experimental method
1) preparation of human muscle hemoglobin solution：Take the human muscle hemoglobin solution of concentration known, with sample dilution buffer (1%BSA,
100mM glycine, 50mM PBS, 150mM NaCl, pH7.4) dilute configuration 25,50,100,300,500ng/ml
Serial human muscle hemoglobin solution.
2) experimental procedure：Planetary reducing motor is uprightly arranged on a precut iron plate, rotating shaft is upward.Made of iron plate straight
Footpath 30mm circular slabs, center punching.Iron plate is horizontal fixed on planetary reducing motor axle, peristaltic pump is installed to iron plate
Center.Planetary reducing motor is connected with potentiometer, then is connected with battery.Peristaltic pump is connected with battery.By sample to be tested
It is fixed on soda liquor container on iron plate., the colloid gold label adsorbed film outside with absorbing membrane pad by myoglobins colloidal-gold detecting-card
On (i.e. liquid adsorption discrete part) inward direction is pasted onto on the outside of iron plate.Peristaltic pump pipette one end is positioned over sample to be tested
In soda liquor container, the other end is fixed on detection stuck point sample groove (colloid gold label adsorbed film).Peristaltic pump pipette feed liquor
Side, which is provided with, can change the three-way cock for inhaling liquid flow path direction.Collaurum is quantified into chromatographic analysis instrument to be fixed on precut iron plate.
Myoglobins colloidal-gold detecting-card is taken, centrifugal rotor plane is pasted and fixed on the side proximal part of colloid gold label adsorbed film
On, peristaltic pump pipette one end is positioned in sample to be tested and soda liquor container, the other end is fixed to detection stuck point sample groove (glue
Body gold mark adsorbed film) on.Peristaltic pump pipette threeway is opened to sample to be checked.Unlatching peristaltic pump, speed governing to 50ul/min,
See that sample liquid to be checked flows out from peristaltic pump pipette point sample groove end, flow on colloid gold label adsorbed film.Open planetary reducing motor,
With potentiometer speed governing to 1000 revs/min, after centrifuging 2 minutes, threeway is turned into cleaning fluid, adjusts planetary reducing motor rotating speed to 2000
Rev/min, wriggling pump speed is adjusted to 150ul/min, and peristaltic pump is closed in centrifugation after 1 minute, adjusts planetary reducing motor rotating speed to 5000
Rev/min, continue centrifugation 30 seconds, close planetary reducing motor.Detection card is removed, is positioned on the quantitative chromatographic analysis instrument of collaurum
Read result.Be 25 to concentration successively, 50,100,300,500ng/ml serial human muscle hemoglobin solution detects.It is real
Test in triplicate, results averaged.
Myoglobins colloidal-gold detecting-card separately is taken, the serial human muscle hemoglobin of 100ul concentration knowns is added dropwise in each card to specifications
Solution, 20 minutes are stood, detection card is positioned on the quantitative chromatographic analysis instrument of collaurum and reads result.Experiment in triplicate, is tied
Fruit is averaged.
3rd, experimental result
Product is detected using existing collaurum immobilon-p, with plane centrifugal separating device of the present invention and existing conventional method
Make comparisons, observe the technology of the present invention and prior art detection and the correlation of sample true value.As a result the technology of the present invention is shown
The correlation coefficient r of detection is 0.998, and the correlation coefficient r of prior art detection is 0.983, P ＜ 0.05, is significantly better than existing skill
The testing result of art, meanwhile, the detection time of prior art detection is 20 minutes, and the technology of the present invention is 3.5 minutes, illustrates this
Inventive technique not only increases the accuracy of prior art detection, while also shortens detection time.
Control experiment (the unit of the plane centrifugal separating device of the present invention of table 1：ng/ml)
Experiment two, the effect detection of the oblique type centrifugal separating device in the China and foreign countries of the embodiment of the present invention 2
1st, experiment material
Small desk centrifuge (i.e. centrifugal device, Eppendorf, Minispin, angle rotor, output speed 1000-10000
Rev/min), thickness 1.5mm iron plates, diameter 8mm iron staffs, (i.e. the sampling pump of sample introduction part, Baoding wound are sharp for miniature peristaltic pump
Pump industry, model BW100), myoglobins colloidal-gold detecting-card (the i.e. one knot of the immobilon-p of immobilon-p and immobilon-p fixator
Structure, Changzhou is rich to hear enlightening Products), (i.e. measuring samples, Sigma-Aldrich products, catalog number are human muscle hemoglobin
F3879-1G), the quantitative chromatographic analysis instrument (i.e. detector, Norway's Skannex products) of collaurum.
2nd, experimental method
The preparation of human muscle hemoglobin solution：Take the human muscle hemoglobin solution of concentration known, with sample dilution buffer (1%BSA,
100mM glycine, 50mM PBS, 150mM NaCl, pH7.4) dilute configuration 25,50,100,300,500ng/ml
Serial human muscle hemoglobin solution.
The long 50mm of iron staff is cut, cutting diameter 150mm round iron plates, iron staff one end is fixed on round iron plate center, iron staff
The other end fixed vertically with compact centrifuge centrifugal rotor axis.Fixed placement miniature peristaltic pump, battery above iron staff plane
And liquid phase container.By myoglobins colloidal-gold detecting-card with absorbing membrane pad outwards, colloid gold label adsorbed film inward direction is in outer
Oblique type fixed placement is in the rotor centrifugal hole of angle.Peristaltic pump pipette one end is positioned in sample to be tested and soda liquor container, another
End is fixed on detection stuck point sample groove (colloid gold label adsorbed film).Peristaltic pump pipette liquid feeding side, which is provided with, can change imbibition
Flow the three-way cock in direction.
Myoglobins colloidal-gold detecting-card is taken, using the side of colloid gold label adsorbed film (i.e. liquid adsorption discrete part) as closely
Heart end is placed in the rotor centrifugal hole of angle, peristaltic pump pipette one end is positioned in sample to be tested and soda liquor container, the other end
It is fixed on detection stuck point sample groove (colloid gold label adsorbed film).Peristaltic pump pipette threeway is opened to sample to be checked.Open compacted
Dynamic pump, speed governing to 50ul/min, see that sample liquid to be checked flows out from peristaltic pump pipette point sample groove end, flow to colloid gold label absorption
On film.Open compact centrifuge, speed governing to 1000 revs/min, centrifugation 2 minutes after, threeway is turned into cleaning fluid, turn down type from
For scheming rotating speed to 2000 revs/min, wriggling pump speed is adjusted to 150ul/min, and centrifugation closes peristaltic pump after 1 minute, turns type centrifugation down
Machine rotating speed continues centrifugation 30 seconds to 5000 revs/min, closes compact centrifuge.Detection card is taken out, is positioned over the quantitative layer of collaurum
Result is read on analysis analyzer.Be 25 to concentration successively, 50,100,300,500ng/ml serial human muscle hemoglobin solution enters
Row detection.Test in triplicate, results averaged.
Myoglobins colloidal-gold detecting-card separately is taken, the serial human muscle hemoglobin of 100ul concentration knowns is added dropwise in each card to specifications
Solution, 20 minutes are stood, detection card is positioned on the quantitative chromatographic analysis instrument of collaurum and reads result.Experiment in triplicate, is tied
Fruit is averaged.
3rd, experimental result
Product is detected using existing collaurum immobilon-p, with oblique type centrifugal separating device and existing conventional method outside the present invention
Make comparisons, observe the technology of the present invention and prior art detection and the correlation of sample true value.As a result the technology of the present invention is shown
The correlation coefficient r of detection is 0.996, and the correlation coefficient r of prior art detection is 0.987, P ＜ 0.05, is significantly better than existing skill
The testing result of art, meanwhile, the detection time of prior art detection is 20 minutes, and the technology of the present invention is 3.5 minutes, illustrates this
Inventive technique not only increases the accuracy of prior art detection, while also shortens detection time.
Control experiment (the unit of the plane centrifugal separating device of the present invention of table 3：ng/ml)
The plane centrifugal separating device Detection results of rotary moving fixing device are carried in experiment three, the embodiment of the present invention 1
1st, experiment material
Planetary reducing motor (i.e. the motor of centrifugal device, 500-5000 revs/min of output speed, power 60w, customization),
Potentiometer, thickness 1.5mm iron plates, miniature peristaltic pump (i.e. sharp pump industry, model BW100 are created in the sampling pump of sample introduction part, Baoding),
Myoglobins colloidal-gold detecting-card (the i.e. integrative-structure of the immobilon-p of immobilon-p and immobilon-p fixator, the production of Changzhou Bo Wendi companies
Product), diameter 3mm iron staffs (i.e. rotary moving fixing device columnar projections), human muscle hemoglobin (i.e. measuring samples,
Sigma-Aldrich products, catalog number F3879-1G), collaurum quantitative chromatographic analysis instrument (i.e. detector, Norway
Skannex products).
2nd, experimental method
The preparation of human muscle hemoglobin solution：Take the human muscle hemoglobin solution of concentration known, with sample dilution buffer (1%BSA,
100mM glycine, 50mM PBS, 150mM NaCl, pH7.4) dilute configuration 25,50,100,300,500ng/ml
Serial human muscle hemoglobin solution.
Long 10mm diameter 3mm iron staffs 5 are cut, are welded to the centrifugation of the plane centrifugal separating device of embodiment 1 vertically
In the outside telecentricity plane of rotor.Myoglobins colloidal-gold detecting-card is taken, punches, will weld in the proximal part close to loading slot
Vertical iron staff insertion myoglobins colloidal-gold detecting-card proximal part hole in, with colloid gold label adsorbed film, (i.e. liquid adsorption is disperseed
Part) side proximal part be fixed in centrifugal rotor plane, peristaltic pump pipette one end is positioned over sample to be tested and cleaning fluid
In container, the other end is fixed on detection stuck point sample groove (colloid gold label adsorbed film).Peristaltic pump pipette threeway is opened to treat
Sample sheet.Peristaltic pump is opened, speed governing to 50ul/min, sees that sample liquid to be checked flows out from peristaltic pump pipette point sample groove end, flow to
On colloid gold label adsorbed film.Planetary reducing motor is opened,, will after centrifuging 2 minutes with potentiometer speed governing to 1000 revs/min
Threeway turns to cleaning fluid, and adjusting planetary reducing motor rotating speed, wriggling pump speed is adjusted to 150ul/min, centrifuges 1 point to 2000 revs/min
Peristaltic pump is closed after clock, adjusts planetary reducing motor rotating speed to continue centrifugation 30 seconds to 5000 revs/min, closes planetary reducing motor.
Detection card is removed, is positioned on the quantitative chromatographic analysis instrument of collaurum and reads result.Successively to concentration be 25,50,100,300,
500ng/ml serial human muscle hemoglobin solution is detected.Test in triplicate, results averaged.
Myoglobins colloidal-gold detecting-card separately is taken, the serial human muscle hemoglobin of 100ul concentration knowns is added dropwise in each card to specifications
Solution, 20 minutes are stood, detection card is positioned on the quantitative chromatographic analysis instrument of collaurum and reads result.Experiment in triplicate, is tied
Fruit is averaged.
3rd, experimental result
Product is detected using existing collaurum immobilon-p, centrifuged with plane of the present invention with rotary moving fixing device
Separator and existing conventional method are made comparisons, and observe the technology of the present invention and prior art detection and the phase of sample true value
Guan Xing.As a result the correlation coefficient r for showing the technology of the present invention detection is 0.998, and the correlation coefficient r of prior art detection is 0.988,
P ＜ 0.05, the testing result of prior art is significantly better than, meanwhile, the detection time of prior art detection is 20 minutes, this hair
Bright technology is 3.5 minutes, illustrates that the technology of the present invention not only increases the accuracy of prior art detection, while also shorten detection
Time.
2 rotary moving of the present invention of table fixes the control experiment (unit of centrifugal separating device：ng/ml)
Claims (16)
1. one kind centrifuges detection means, it is characterised in that：It include sample introduction part, immobilon-p, centrifugal device and
Detector；
The centrifugal device is included by the centrifugal rotor and support base of motor driving, and the centrifugal rotor is with described
Support base is support；
The sample introduction part is located at the middle part of the centrifugal rotor, and is connected with centrifugal rotor；The sample introduction part bag
Include liquid phase storage device and sample feeding pipe；The liquid phase storage device is connected with the sample feeding pipe；
The immobilon-p is connected on the centrifugal rotor with the sample feeding pipe；
The detector is located at the one or both sides of the immobilon-p.
2. centrifugation detection means according to claim 1, it is characterised in that：The immobilon-p is placed in solid phase
In film apparatus for placing, and the immobilon-p apparatus for placing is on the centrifugal rotor；
The immobilon-p and the centrifugal rotor are dismountable structure.
3. centrifugation detection means according to claim 2, it is characterised in that：The immobilon-p apparatus for placing
Including rotary moving fixing device and/or flute profile squash type fixing device.
4. the centrifugation detection means according to any one of claim 1-3, it is characterised in that：The solid phase
Film is fixed by immobilon-p fixator, and the immobilon-p fixator is selected from immobilon-p support egative film sample part, lateral flow examination
Paper slip buckle sample part and hyalomitome embed at least one of formula part up and down.
5. centrifugation detection means according to claim 4, it is characterised in that：The hyalomitome embeds up and down
Formula part is covered for the both sides up and down of the immobilon-p by hard transparent material, the hard of the immobilon-p respective side
The area of transparent material is more than or equal to the area of the immobilon-p.
6. the centrifugation detection means according to any one of claim 1-5, it is characterised in that：The solid phase
Film is additionally provided with the one end being connected with the sample feeding pipe is attached thereto logical liquid adsorption discrete part, the immobilon-p in
The distal end of the centrifugal rotor is provided with and is attached thereto logical liquid collecting member, the liquid adsorption discrete part and institute
Sample feeding pipe is stated to be connected.
7. the centrifugation detection means according to any one of claim 1-6, it is characterised in that：The sample introduction
Part includes sampling pump, and the sampling pump drives the liquid in the liquid phase storage device to enter the sample feeding pipe.
8. the centrifugation detection means according to any one of claim 7, it is characterised in that：The liquid phase storage
Cryopreservation device includes measuring samples storage device and detection phase storage device；
The measuring samples storage device and the detection phase storage device are connected with the sample feeding pipe, and by institute
State sampling pump driving.
9. the centrifugation detection means according to claim 7 or 8, it is characterised in that：The liquid phase storage dress
Putting also includes cleaning fluid storage device, and the cleaning fluid storage device is connected with the sample feeding pipe, and by the sample introduction
Pump drives.
10. the centrifugation detection means according to any one of claim 1-9, it is characterised in that：It is described from
Heart rotor is using plane or by the outward-dipping type in center；
The centrifugal device is provided with a shell.
11. the centrifugation detection means according to any one of claim 4-10, it is characterised in that：The rotation
Transfer dynamic formula fixing device is the attachment means of the columnar projections set on the centrifugal rotor, and the immobilon-p is fixed
Device is provided with the poroid part to match with the attachment means of the columnar projections.
12. the centrifugation detection means according to any one of claim 1-11, it is characterised in that：It is described from
Center device, the sample introduction part and the detector are equipped with presetting apparatus.
13. the centrifugation detection means according to any one of claim 1-12, it is characterised in that：It is described enter
Sample pipe is connected with the immobilon-p using dismountable；
The liquid adsorption discrete part is connected with the sample feeding pipe using dismountable.
14. the centrifugation detection means according to any one of claim 1-13, it is characterised in that：It is described solid
The material of phase film uses any of nitrocellulose filter, polyvinylidene fluoride film, nylon membrane and DEAE cellulose membranes,
And the one or both sides of the immobilon-p are provided with backing；
The liquid adsorption discrete part includes colloid gold label adsorbed film, fluorescent labeled antibody adsorbed film, chemiluminescence
Mark at least one of adsorbed film, poly ester fiber dispersion membrane and glass fibre dispersion membrane；
The detector includes any of detector of absorbance, fluorescence, chemiluminescence and image processing.
15. the centrifugation detection means stated according to any one of claim 1-14, it is characterised in that：The centrifugation
The plane interlude of rotor is provided with hole and/or provided with transparent component, and the hole and/or the transparent component make it is described solid
Phase film is directly exposed to the detector.
16. application of the centrifugation detection means in immunoassay product detection any one of claim 1-15,
Specifically, the immunoassay product includes at least one of antigen, antibody, immunocyte and chemical composition.
Priority Applications (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610330378.7A CN107402299A (en) | 2016-05-18 | 2016-05-18 | One kind centrifuges detection means |
| JP2018547942A JP2019507887A (en) | 2016-03-14 | 2016-06-23 | Centrifugation detection method and apparatus |
| EP16894072.4A EP3431990A4 (en) | 2016-03-14 | 2016-06-23 | Centrifugation detection method and device |
| PCT/CN2016/086842 WO2017156910A1 (en) | 2016-03-14 | 2016-06-23 | Centrifugation detection method and device |
| US16/124,858 US20180364227A1 (en) | 2016-03-14 | 2018-09-07 | Detection method with centrifuge isolation and detection device thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610330378.7A CN107402299A (en) | 2016-05-18 | 2016-05-18 | One kind centrifuges detection means |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107402299A true CN107402299A (en) | 2017-11-28 |
Family
ID=60394523
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610330378.7A Withdrawn CN107402299A (en) | 2016-03-14 | 2016-05-18 | One kind centrifuges detection means |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107402299A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107688089A (en) * | 2016-08-03 | 2018-02-13 | 北京康华源科技发展有限公司 | One kind centrifuges detection means and application thereof |
| CN111289763A (en) * | 2020-03-03 | 2020-06-16 | 江苏省肿瘤防治研究所（江苏省肿瘤医院） | Antibody sampling analyzer and use method thereof |
-
2016
- 2016-05-18 CN CN201610330378.7A patent/CN107402299A/en not_active Withdrawn
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107688089A (en) * | 2016-08-03 | 2018-02-13 | 北京康华源科技发展有限公司 | One kind centrifuges detection means and application thereof |
| CN107688089B (en) * | 2016-08-03 | 2020-05-15 | 北京康华源科技发展有限公司 | Centrifugal separation detection device and application thereof |
| CN111289763A (en) * | 2020-03-03 | 2020-06-16 | 江苏省肿瘤防治研究所（江苏省肿瘤医院） | Antibody sampling analyzer and use method thereof |
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Application publication date: 20171128 |