CN208894245U - The micro-fluidic chip of rapid quantitative detection PLGF and sFLT-1 - Google Patents
The micro-fluidic chip of rapid quantitative detection PLGF and sFLT-1 Download PDFInfo
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- CN208894245U CN208894245U CN201820992592.3U CN201820992592U CN208894245U CN 208894245 U CN208894245 U CN 208894245U CN 201820992592 U CN201820992592 U CN 201820992592U CN 208894245 U CN208894245 U CN 208894245U
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Abstract
The utility model discloses the micro-fluidic chips of rapid quantitative detection PLGF and sFLT-1, including chip substrate and chip cover board, chip substrate is with chip cover board by being bonded, 2 microfluidic channels being provided symmetrically on chip substrate, 2 microfluidic channels include sample application zone, reaction zone, detection zone, waste, reaction zone is coated with the PLGF antibody I of fluorescent microsphere label or the sFLT-1 antibody I of fluorescent microsphere label, the fixed PLGF antibody I I or sFLT-1 antibody I I for being coated with different epitopes in detection zone；The utility model has the advantages that high sensitivity, favorable repeatability, required sample size is few, detection time is short, it is easy to operate, detection is full-automatic, can detect simultaneously to PLGF and sFLT-1, doctor can quickly be helped to predict preeclampsia risk, EARLY RECOGNITION is carried out to people at highest risk and is intervened, to ensure gestational period Material and infant safe.
Description
Technical field
The utility model relates to field of medical examination, and in particular to rapid quantitative detection PLGF's and sFLT-1 is micro-fluidic
Chip.
Background technique
Soluble vascular endothelial growth factor receptor1 (sVEGFR-1), also referred to as sFlt-1
(sFLT-1), it is initially found in human umbilical vein endothelial cell, is a kind of glycoprotein with tyrosine kinase activity；Placenta
Growth factor (PLGF) is the angiogenesis factor by placenta secretion, can promote the proliferation of vascular endothelial cell, is a kind of master
The angiogenesis factor wanted, and increase the permeability of endothelial cell；As one of the gestational period distinctive severe complication, before eclampsia
Phase incidence is about the 3-5% of all gestation, and 15% premature labor and 42% pregnant woman's death are caused by preeclampsia, strong to mother and baby
Kang Zaocheng is seriously threatened.Timely, accurate diagnosis is carried out to the pregnant woman for having preeclampsia risk in morbidity early stage and is conducive to clinic
Patient is monitored closely, carries out effectively nursing in time to control the state of an illness, extend pregnant week, for reducing Attack of Preeclampsia rate
It is of great significance with the death rate, guarantee Material and infant safe.Currently, existing numerous studies prove the change of sFLT-1 and PLGF concentration
Obviously earlier than Attack of Preeclampsia, sFLT-1/PLGF ratio can preferably reflect the growing state of placenta blood vessel, assess
On the basis of albuminuria and blood pressure, joint-detection sFLT-1/PLGF ratio can help clinic that preeclampsia trouble is better anticipated
Sick risk helps doctor to carry out EARLY RECOGNITION to people at highest risk and intervenes, to ensure gestational period Material and infant safe.
Microfluidic chip technology is bases such as biology, chemistry, the sample preparation of medical analysis process, reaction, separation, detections
This operating unit is integrated on the chip of one piece of micro-meter scale, is automatically performed analysis overall process；Operation can be greatly simplified
Process, reduces sample and reagent consumes, and does not need to be equipped with expensive instrument, makes it possible that scene is detected immediately, tool
There are foreseeable huge economic value and social value；Therefore, a kind of micro-fluidic chip for detecting PLGF and sFLT-1 is established
Technology platform has a very important significance.
Utility model content
The purpose of this utility model is to provide the micro-fluidic chips of rapid quantitative detection PLGF and sFLT-1, can be quick
It helps doctor to predict preeclampsia risk, EARLY RECOGNITION is carried out to people at highest risk and intervenes, to ensure gestational period mother and baby
Safety.
To achieve the above object, the utility model provides the following technical solutions: rapid quantitative detection PLGF's and sFLT-1
Micro-fluidic chip, including chip substrate and chip cover board, chip substrate, by being bonded, are wrapped with chip cover board on chip substrate
Include symmetrically arranged 2 microfluidic channels, respectively the first microfluidic channel, the second microfluidic channel, the first microfluidic channel,
Second microfluidic channel includes the sample application zone being sequentially communicated, reaction zone, detection zone, waste, and the first microfluidic channel is used for
Quantitative detection PLGF, the reaction zone in the first microfluidic channel are coated with the PLGF antibody I of fluorescent microsphere label, and first is micro-fluidic
Detection zone in channel is coated with the PLGF antibody I I of fluorescent microsphere label, and the second microfluidic channel is used for quantitative detection sFLT-
1, the reaction zone in the second microfluidic channel is coated with the sFLT-1 antibody I of fluorescent microsphere label, in the second microfluidic channel
Detection zone is coated with the sFLT-1 antibody I I of fluorescent microsphere label；Chip cover board be equipped with well, the position of well with plus
Sample area is corresponding, and well is located above sample application zone；
The chip substrate, chip cover board section be circular configuration；
The microfluidic channel is arranged along the radial direction of chip substrate, the distance of 2 microfluidic channels to chip substrate
It is equal；
The sample application zone is close to the center of chip substrate；
The sample application zone is connected with reaction zone by first passage, filter pad is equipped in first passage, filter pad is complete
Blood seperation film, macropore can retain red blood cell, the leucocyte in whole blood by physics mode in film, and blood plasma is small from film downstream face
Hole outflow can read the blood plasma in separation whole blood without haemolysis fastly, avoid influence of the red blood cell to testing result；The reaction
Area is connected with detection zone by second channel, and current limliting column is equipped in second channel, and fluid can be effectively controlled second in current limliting column
Time in channel, to make fluid to be measured and antibody response, detection zone is connected with waste by third channel sufficiently, completely
It is logical, gag lever post is equipped in third channel, for gag lever post close to detection zone, gag lever post can stop the waste liquid in waste to return to detection
Area, gag lever post are equipped with detent projection towards the side of waste, convenient for drainage, the fluid of detection zone are enabled smoothly to flow into waste liquid
Area；
The current limliting column includes the first restricted section of integrated design, the second restricted section, third restricted section, the second current limliting section
Between the first restricted section, third restricted section, the first restricted section, third restricted section section be string configuration, the second current limliting
The section of section is s shape structure, and the quantity of current limliting column is 2, and 2 current limliting columns are arranged in parallel, shape between second channel and current limliting column
At diversion trench；
The section of the sample application zone be isosceles trapezoidal structure, reaction zone, detection zone section be circular configuration, waste
Section is isosceles triangular structure.
The micro-fluidic chip of above-mentioned a kind of rapid quantitative detection PLGF and sFLT-1, wherein the chip substrate, chip lid
Plate selects one of dimethyl silicone polymer (PDMS), polymethyl methacrylate, polytetrafluoroethylene (PTFE).
The micro-fluidic chip of above-mentioned a kind of rapid quantitative detection PLGF and sFLT-1, wherein the volume of the waste is big
In the sum of sample application zone, reaction zone, the total volume of detection zone.
The utility model also provides a kind of method for preparing above-mentioned micro-fluidic chip, includes the following steps:
(1) filter pad is fixed in first passage to the side for leaning on sample adding area；
(2) fluorescent microsphere is washed with 0.05mol/L MES buffer (pH=7.2), after washing, be added carbodiimide and
N-hydroxysuccinimide reacts at room temperature 2h, obtains fluorescent microsphere solution, antibody I is dissolved in the PBS buffer solution of 0.05mol/L
(pH=7.2), antibody I solution is obtained, fluorescent microsphere solution is added in antibody I solution, reacts at room temperature 2h, centrifugal filtration is used
The PBS buffer solution (pH=7.2) of 0.01mol/L is washed 3-5 times, obtains the antibody I of fluorescent microsphere label, fluorescent microsphere is marked
Antibody I drop coating in reaction zone；
(3) antibody I I fixation is coated in detection zone.
Testing principle: adding sample application zone for whole blood sample from well, and under the driving of centrifugal force, whole blood sample passes through
Filtered points in first passage remove red blood cell, leucocyte, subsequently enter reaction zone；Antigen and fluorescent microsphere mark in sample
The antibody response of note is formed, and Ag-Ab-fluorescent microsphere compound is formed, anti-in reaction zone under the driving of centrifugal force
Antigen-antibody I- fluorescent microsphere compound enters detection zone from the diversion trench of second channel, and Ag-Ab I- fluorescent microsphere is multiple
It closes the antibody I I that object and detection zone are fixed and specific immune response occurs, it is double to form fluorescent microsphere-antibody I-Ag-Ab II
Antibody sandwich compound, and it is fixed on detection zone；Under the driving of centrifugal force, the sample that do not fix enters waste, passes through
Cleaning solution is added in well, cleans microfluidic channel, finally detects micro-fluidic chip by fluorescence detector.
The utility model has the utility model has the advantages that the utility model provides a kind of rapid quantitative detection PLGF and sFLT-1
Micro-fluidic chip, high sensitivity, favorable repeatability, required sample size is few, and detection time is short, easy to operate, detection entirely from
It is dynamic, PLGF and sFLT-1 can be detected simultaneously, operating process can be greatly simplified, sample is reduced and reagent disappears
It consumes, and does not need to be equipped with expensive instrument, make it possible that scene is detected immediately, there is huge economic value and society's valence
Value；It can quickly help doctor to predict preeclampsia risk simultaneously, EARLY RECOGNITION is carried out to people at highest risk and intervene, thus
Ensure gestational period Material and infant safe.
Detailed description of the invention
Fig. 1 is the utility model chip substrate structure schematic diagram；
Fig. 2 is microfluidic channel structural schematic diagram；
Fig. 3 is enlarged drawing at A in Fig. 2；
In figure: 1- chip substrate, the first microfluidic channel of 2-, the second microfluidic channel of 3-, the sample application zone 4-, 5- reaction zone,
6- detection zone, 7- waste, 8- first passage, 9- second channel, 10- third channel, 11- filter pad, 12- current limliting column, 13-
First restricted section, the second restricted section of 14-, 15- third restricted section, 16- gag lever post.
Specific embodiment
The following will be combined with the drawings in the embodiments of the present invention, carries out the technical scheme in the embodiment of the utility model
Clearly and completely describe.
Embodiment 1
The present embodiment illustrates the structure of micro-fluidic chip；
As shown in Figs. 1-2, the micro-fluidic chip of rapid quantitative detection PLGF and sFLT-1, including chip substrate and chip lid
Plate, chip substrate, by being bonded, include symmetrically arranged 2 microfluidic channels on chip substrate, respectively with chip cover board
For the first microfluidic channel, the second microfluidic channel, the first microfluidic channel, the second microfluidic channel include being sequentially communicated
Sample application zone, reaction zone, detection zone, waste, the first microfluidic channel is used for quantitative detection PLGF, in the first microfluidic channel
Reaction zone is coated with the PLGF antibody I of fluorescent microsphere label, and the detection zone in the first microfluidic channel is coated with fluorescent microsphere mark
The PLGF antibody I I of note, the second microfluidic channel are used for quantitative detection sFLT-1, the reaction zone coating in the second microfluidic channel
There is the sFLT-1 antibody I of fluorescent microsphere label, the detection zone in the second microfluidic channel is coated with the sFLT- of fluorescent microsphere label
1 antibody I I；Chip cover board is equipped with well, and the position of well is corresponding with sample application zone, and well is located above sample application zone；
The chip substrate 1, chip cover board section be circular configuration；
The microfluidic channel is arranged along the radial direction of chip substrate, the distance of 2 microfluidic channels to chip substrate
It is equal；
The sample application zone is close to the center of chip substrate；
The sample application zone is connected with reaction zone by first passage 8, and filter pad 11 is equipped in first passage, and filter pad is
Whole blood seperation film, macropore can retain red blood cell, the leucocyte in whole blood by physics mode in film, and blood plasma is from film downstream face
Aperture outflow can read the blood plasma in separation whole blood without haemolysis fastly, avoid influence of the red blood cell to testing result；It is described anti-
It answers area to be connected with detection zone by second channel, current limliting column 12 is equipped in second channel 9, current limliting column can be effectively controlled fluid and exist
Time in second channel, to make fluid to be measured and antibody response, detection zone and waste pass through third channel sufficiently, completely
10 are connected, and gag lever post is equipped in third channel, and gag lever post can stop the waste liquid in waste to return close to detection zone, gag lever post
Detection zone is returned, gag lever post 16 is equipped with detent projection towards the side of waste, convenient for drainage, enables the fluid of detection zone smooth
Flow into waste；
First restricted section 13, second restricted section 14, third restricted section 15 of the current limliting column 12 including integrated design, second
Restricted section between the first restricted section, third restricted section, the first restricted section, third restricted section section be string configuration,
The section of second restricted section is s shape structure, and the quantity of current limliting column is 2, and 2 current limliting columns are arranged in parallel, second channel and current limliting
Diversion trench is formed between column；
The section of the sample application zone be isosceles trapezoidal structure, reaction zone, detection zone section be circular configuration, waste
Section is isosceles triangular structure.
The micro-fluidic chip of above-mentioned a kind of rapid quantitative detection PLGF and sFLT-1, wherein the chip substrate, chip lid
Plate selects one of dimethyl silicone polymer (PDMS), polymethyl methacrylate, polytetrafluoroethylene (PTFE).
The micro-fluidic chip of above-mentioned a kind of rapid quantitative detection PLGF and sFLT-1, wherein the volume of the waste is big
In the sum of sample application zone, reaction zone, the total volume of detection zone.
Embodiment 2
The preparation method of the micro-fluidic chip of quantitative detection PLGF is illustrated in the present embodiment；
Chip substrate is equipped with the first microfluidic channel, the second microfluidic channel, wherein the first microfluidic channel is for fixed
Amount detection PLGF, specifically comprises the following steps:
(1) filter pad is fixed in first passage to the side for leaning on sample adding area；
(2) fluorescent microsphere is washed with 0.05mol/L MES buffer (pH=7.2), after washing, be added carbodiimide and
N-hydroxysuccinimide reacts at room temperature 2h, obtains fluorescent microsphere solution, the PBS that PLGF antibody I is dissolved in 0.05mol/L is delayed
Fliud flushing (pH=7.2) obtains PLGF antibody I solution, and fluorescent microsphere solution is added in PLGF antibody I solution, reacts at room temperature 2h,
Centrifugal filtration is washed 3-5 times with the PBS buffer solution (pH=7.2) of 0.01mol/L, obtains the PLGF antibody of fluorescent microsphere label
I, by the PLGF antibody I drop coating of fluorescent microsphere label in reaction zone；
(3) PLGF antibody I I fixation is coated in detection zone.
All reagents of the utility model are fixed in chip in advance, eliminate the use of a large amount of test tubes of conventional reagents box,
And sampling operation repeatedly is save, save manpower and material resources；
When detection: whole blood sample being added sample application zone from well, under the driving of centrifugal force, whole blood sample is by the
Filtered points in one channel remove red blood cell, leucocyte, subsequently enter reaction zone；Antigen and fluorescent microsphere in sample mark
PLGF antibody response formed, formed antigen-PLGF antibody-fluorescent microsphere compound, under the driving of centrifugal force, in reaction zone
Antigen-PLGF antibody I-fluorescent microsphere compound from the diversion trench of second channel enter detection zone, antigen-PLGF antibody I-
Specific immune response occurs for the PLGF antibody I I that fluorescent microsphere compound and detection zone are fixed, and it is anti-to form fluorescent microsphere-PLGF
The double-antibody sandwich compound of body I- antigen-PLGF antibody I I, and it is fixed on detection zone；It is not solid under the driving of centrifugal force
Fixed sample enters waste, and cleaning solution is added by well, cleans microfluidic channel, finally passes through micro-fluidic chip glimmering
Optical detector is detected, and the luminous intensity of fluorescent microsphere is detected, and fluorescence intensity is in correlation with testing concentration.
Embodiment 3
The preparation method of the micro-fluidic chip of quantitative detection sFLT-1 is illustrated in the present embodiment；
Chip substrate is equipped with the first microfluidic channel, the second microfluidic channel, wherein the second microfluidic channel is for fixed
Amount detection sFLT-1, specifically comprises the following steps:
(1) filter pad is fixed in first passage to the side for leaning on sample adding area；
(2) fluorescent microsphere is washed with 0.05mol/L MES buffer (pH=7.2), after washing, be added carbodiimide and
N-hydroxysuccinimide reacts at room temperature 2h, obtains fluorescent microsphere solution, sFLT-1 antibody I is dissolved in the PBS of 0.05mol/L
Buffer (pH=7.2) obtains sFLT-1 antibody I solution, fluorescent microsphere solution is added in sFLT-1 antibody I solution, room temperature
2h is reacted, centrifugal filtration is washed 3-5 times with the PBS buffer solution (pH=7.2) of 0.01mol/L, obtains fluorescent microsphere label
SFLT-1 antibody I, by the sFLT-1 antibody I drop coating of fluorescent microsphere label in reaction zone；
(3) sFLT-1 antibody I I fixation is coated in detection zone.
When detection: whole blood sample being added sample application zone from well, under the driving of centrifugal force, whole blood sample is by the
Filtered points in one channel remove red blood cell, leucocyte, subsequently enter reaction zone；Antigen and fluorescent microsphere in sample mark
SFLT-1 antibody response formed, formed antigen-sFLT-1 antibody-fluorescent microsphere compound, under the driving of centrifugal force, reaction
Antigen-sFLT-1 antibody I-fluorescent microsphere compound in area enters detection zone, antigen-from the diversion trench of second channel
Specific immune response occurs for the sFLT-1 antibody I I that sFLT-1 antibody I-fluorescent microsphere compound and detection zone are fixed, and is formed
Fluorescent microsphere-sFLT-1 antibody I-antigen-sFLT-1 antibody I I double-antibody sandwich compound, and it is fixed on detection zone；From
Under the driving of mental and physical efforts, the sample that do not fix enters waste, and cleaning solution is added by well, cleans microfluidic channel, most
Micro-fluidic chip is detected by fluorescence detector afterwards, detects the luminous intensity of fluorescent microsphere, fluorescence intensity and determinand
Concentration is in correlation.
By the sFLT-1 concentration to be converted in embodiment 3 by fluorescence intensity and implement in 2 through fluorescence intensity conversion
PLGF concentration conversion can help clinic that preeclampsia is better anticipated at sFLT-1/PLGF ratio, sFLT-1/PLGF ratio
Risk helps doctor to carry out EARLY RECOGNITION to people at highest risk and intervenes, to ensure gestational period Material and infant safe.
While there has been shown and described that the embodiments of the present invention, for the ordinary skill in the art,
It is understood that these embodiments can be carried out with a variety of variations in the case where not departing from the principles of the present invention and spirit, repaired
Change, replacement and variant, the scope of the utility model is defined by the appended claims and the equivalents thereof.
Claims (3)
1. the micro-fluidic chip of rapid quantitative detection PLGF and sFLT-1, which is characterized in that including chip substrate and chip cover board,
Chip substrate and chip cover board include symmetrically arranged 2 microfluidic channels on chip substrate by being bonded, and respectively the
One microfluidic channel, the second microfluidic channel, the first microfluidic channel, the second microfluidic channel include the sample-adding being sequentially communicated
Area, reaction zone, detection zone, waste, the first microfluidic channel are used for quantitative detection PLGF, the reaction in the first microfluidic channel
Area is coated with the PLGF antibody I of fluorescent microsphere label, and the detection zone in the first microfluidic channel is coated with fluorescent microsphere label
PLGF antibody I I, the second microfluidic channel are used for quantitative detection sFLT-1, and the reaction zone in the second microfluidic channel is coated with glimmering
The sFLT-1 antibody I of light microballoon label, the sFLT-1 that the detection zone in the second microfluidic channel is coated with fluorescent microsphere label are anti-
Body II；Chip cover board is equipped with well, and the position of well is corresponding with sample application zone, and well is located above sample application zone；
The chip substrate, chip cover board section be circular configuration；
The microfluidic channel is arranged along the radial direction of chip substrate, 2 microfluidic channel being equidistant to chip substrate；
The sample application zone is close to the center of chip substrate；The sample application zone is connected with reaction zone by first passage, and first is logical
Filter pad is equipped in road, filter pad is whole blood seperation film, and the reaction zone is connected with detection zone by second channel, and second is logical
Current limliting column is equipped in road, detection zone is connected with waste by third channel, and gag lever post is equipped in third channel, and gag lever post leans on
Nearly detection zone, gag lever post are equipped with detent projection towards the side of waste；
The current limliting column includes the first restricted section of integrated design, the second restricted section, third restricted section, and the second restricted section is located at the
Between one restricted section, third restricted section, the first restricted section, third restricted section section be string configuration, the second restricted section
Section is s shape structure, and the quantity of current limliting column is 2, and 2 current limliting columns are arranged in parallel, and are formed and are led between second channel and current limliting column
Chute；
The section of the sample application zone be isosceles trapezoidal structure, reaction zone, detection zone section be circular configuration, the section of waste
For isosceles triangular structure.
2. the micro-fluidic chip of rapid quantitative detection PLGF and sFLT-1 as described in claim 1, which is characterized in that the core
Plate base, chip cover board select one of dimethyl silicone polymer (PDMS), polymethyl methacrylate, polytetrafluoroethylene (PTFE).
3. the micro-fluidic chip of rapid quantitative detection PLGF and sFLT-1 as described in claim 1, which is characterized in that described useless
The volume of liquid zone is greater than the sum of sample application zone, reaction zone, the total volume of detection zone.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201820992592.3U CN208894245U (en) | 2018-06-26 | 2018-06-26 | The micro-fluidic chip of rapid quantitative detection PLGF and sFLT-1 |
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| CN201820992592.3U CN208894245U (en) | 2018-06-26 | 2018-06-26 | The micro-fluidic chip of rapid quantitative detection PLGF and sFLT-1 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111812329A (en) * | 2020-07-28 | 2020-10-23 | 山东圣剑医学研究有限公司 | Antibody chip kit for quantitatively detecting multiple tumor markers |
| CN114405568A (en) * | 2022-03-04 | 2022-04-29 | 南开大学 | Self-driven micro-fluidic chip |
-
2018
- 2018-06-26 CN CN201820992592.3U patent/CN208894245U/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111812329A (en) * | 2020-07-28 | 2020-10-23 | 山东圣剑医学研究有限公司 | Antibody chip kit for quantitatively detecting multiple tumor markers |
| CN114405568A (en) * | 2022-03-04 | 2022-04-29 | 南开大学 | Self-driven micro-fluidic chip |
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