CN215179669U

CN215179669U - Micro-fluidic chip and whole blood separation and blood coagulation medicine real-time detection device

Info

Publication number: CN215179669U
Application number: CN202120473977.0U
Authority: CN
Inventors: 丁涛; 周星贝
Original assignee: Individual
Current assignee: Individual
Priority date: 2021-03-04
Filing date: 2021-03-04
Publication date: 2021-12-14
Anticipated expiration: 2031-03-04

Abstract

The utility model provides a micro-fluidic chip, including dividing the blood runner and detecting the runner, divide the blood runner to be helical structure, divide the feed liquor end of blood runner to be located helical structure's inner circle, divide the play liquid end of blood runner to be located helical structure's outermost circle, divide the play liquid end branch of blood runner to be centrifugal outside runner and centripetal inboard runner, outside runner connection detection runner. The device comprises a blood collector, a power pump, an FXa injection pump, a fluorescent reagent injection pump, a fluorescence measuring instrument, a data processing unit and the microfluidic chip. The utility model can effectively separate whole blood for real-time detection, thereby improving the timeliness; the detection method adopts an anti-FXa method to specifically measure the FXa inhibitor in blood, directly reflects the blood concentration and improves the accuracy; the quantitative method and the fluorescence quantitative method are used, so that the detection sensitivity is improved.

Description

Micro-fluidic chip and whole blood separation and blood coagulation medicine real-time detection device

Technical Field

The utility model relates to a blood detection device technical field, more specifically relates to a micro-fluidic chip and a whole blood separation and blood coagulation medicine real-time detection device.

Background

Blood is one of the most common samples in clinical examination, and various physiological and pathological information of a testee can be obtained from the blood by various detection means such as biochemistry, immunity and the like, so that a basis is provided for clinical diagnosis and treatment. At present, most of tests aiming at blood need to separate blood cells from liquid components in a sample pretreatment stage and then carry out subsequent tests on enriched blood cells or serum and plasma. The traditional blood cell separation methods include centrifugation, filtration, salting out and the like, and the methods generally take a long time and are complicated to operate.

FXa inhibitors are a common class of clinical anticoagulants. In the course of treatment, excessive administration can lead to bleeding, and insufficient anticoagulation often causes thrombosis, thereby causing serious consequences. Therefore, close monitoring of the coagulation status of patients is required in the application of FXa inhibitors, especially for a part of people with undefined drug metabolism, such as renal insufficiency patients, obese or wasting people, children, pregnant women, the elderly, etc. The concentration of FXa inhibitors in the blood is a very important therapeutic efficacy indicating tool. The currently clinically common anticoagulation monitoring items are Activated Partial Thromboplastin Time (APTT) and Activated Clotting Time (ACT). APTT and ACT do not directly measure drug concentration in blood, and the results are prone to error. And due to limitations in the measurement principle, these methods are not applicable to blood samples using large doses of FXa inhibitors. The anti-Xa method is a gold standard for measuring FXa inhibitors and is currently being popularized and applied gradually as an alternative method to ACT and APTT. However, the cellular components in the blood interfere with the results of the anti-Xa assay, so the whole blood sample needs to be pretreated by cell separation before the assay, the treatment process is prolonged, and the timeliness is insufficient. The current technical means needs manual blood drawing, inspection and other steps for single measurement. Although there are also devices or methods in the current state of the art that enable immediate detection or bedside detection of the anticoagulation function, for example chinese patent CN111094990A discloses a method and a device for detecting anticoagulants in plasma and whole blood by adding coagulation factors to portions of said blood sample, each portion receiving a different concentration of said coagulation factor; measuring clot formation for each portion of the sample; and determining the response of blood clot formation to the concentration of the coagulation factor. It detects anticoagulants by the relationship of clot formation, measured by imaging, to coagulation factors; the measurement process is complex, the detection sensitivity is not high, the time consumption is long, the result cannot be quickly obtained, and the timeliness is insufficient; and blood samples using a large dose of FXa inhibitor cannot be accurately measured, and the application range is narrow.

SUMMERY OF THE UTILITY MODEL

The utility model discloses an overcome among the above-mentioned background art technical means measurement process comparatively complicated, unable accurate quantitative analysis to length consuming time, unable quick acquisition result, the not enough problem of ageing provide a micro-fluidic chip and a whole blood separation and blood coagulation medicine real-time detection device. The utility model discloses can effectively separate whole blood, detectivity is high, can carry out real-time detection fast effectively.

In order to solve the technical problem, the utility model discloses a technical scheme is: the utility model provides a micro-fluidic chip for whole blood separation and detection, is including dividing the blood flow way and detecting flow way, divide the blood flow way to be helical structure, the feed liquor end that divides the blood flow way is located helical structure's inner circle, the play liquid end that divides the blood flow way is located helical structure's outermost circle, and the play liquid end fork that divides the blood flow way is centrifugal outside runner and centripetal inboard runner, the outside runner is connected detecting flow way.

Furthermore, the detection flow channel comprises a main flow channel and a first branch flow channel and a second branch flow channel which are used for injecting different detection reagents respectively, and the first branch flow channel and the second branch flow channel are communicated with one end of the main flow channel, which is close to the liquid outlets of the blood distribution flow channels, respectively.

Furthermore, the main runner comprises a buffer runner and a direct-current runner for detecting the fluorescence measuring instrument, the outer side runner, the first branch runner and the second branch runner converge to the inlet of the buffer runner, and the outlet of the buffer runner is connected with one end of the direct-current runner.

Preferably, the buffer flow channel has a serpentine structure. The buffer channel is used for fully mixing the FXa and the fluorescent reagent with the plasma.

Still provide a whole blood separation and coagulation medicine real-time detection device, including blood collector, power pump, FXa syringe pump, fluorescence reagent syringe pump, fluorescence measurement appearance, data processing unit and foretell micro-fluidic chip, power pump one end is connected blood collector, the other end are connected divide the inlet port of blood flow way, first runner is connected FXa syringe pump, second runner are connected fluorescence reagent syringe pump, fluorescence measurement appearance is located the end of detection flow way, data processing unit with fluorescence measurement appearance connects.

Further, the observation area of the fluorescence measuring instrument is aligned with the straight flow channel.

Further, the blood collector is a venous cannula.

Preferably, the fluorescence detection excitation wavelength of the fluorescence measuring instrument is 350nm, and the emission wavelength is 450 nm.

Preferably, the power pump is a peristaltic pump.

Preferably, the fluorescence measuring instrument is an Axio Imager 2 fluorescence measuring instrument.

Compared with the prior art, the beneficial effects are:

1. the utility model can quickly and effectively separate whole blood, perform real-time anticoagulation detection, shorten the time for measuring the concentration of FXa inhibitor to within 30 minutes, and improve the timeliness; the detection method adopts an anti-FXa method to specifically measure the FXa inhibitor in blood, directly reflects the blood concentration and improves the accuracy; the quantitative method and the fluorescence quantitative method are used, so that the detection sensitivity is improved.

2. The utility model can reliably measure the FXa inhibitor concentration within the range of 0-1.5IU/ml, and broadens the measurable range of the concentration; and the test needs less blood, only about 2ml of blood is needed for continuous monitoring for 24 hours, and the damage to the patient is small.

3. The utility model discloses still optimize to the sample link, adopt the automatic sample of mechanical pump, reduce manual operation, concentration measurement data can be once obtained every 15s, has realized online real-time supervision.

Drawings

FIG. 1 is a schematic structural view of example 1.

FIG. 2 is a schematic structural view of example 2.

Detailed Description

The drawings are for illustrative purposes only and are not to be construed as limiting the patent; for the purpose of better illustrating the embodiments, certain features of the drawings may be omitted, enlarged or reduced, and do not represent the size of an actual product; it will be understood by those skilled in the art that certain well-known structures in the drawings and descriptions thereof may be omitted. The positional relationships depicted in the drawings are for illustrative purposes only and are not to be construed as limiting the present patent.

The same or similar reference numerals in the drawings of the embodiments of the present invention correspond to the same or similar parts; in the description of the present invention, it should be understood that if there are the terms "upper", "lower", "left", "right", "long", "short", etc. indicating the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, it is only for convenience of description and simplification of description, but it is not intended to indicate or imply that the device or element referred to must have a specific orientation, be constructed in a specific orientation, and be operated, and therefore the terms describing the positional relationship in the drawings are only for illustrative purposes and are not to be construed as limiting the present patent, and those skilled in the art will understand the specific meaning of the terms according to their specific circumstances.

The technical solution of the present invention is further described in detail by the following specific embodiments in combination with the accompanying drawings:

example 1

As shown in fig. 1, the microfluidic chip is mainly used for whole blood separation and detection, and comprises a blood separation channel 1 and a detection channel 2, wherein the blood separation channel 1 is of a spiral structure, a liquid inlet end 3 of the blood separation channel 1 is positioned at the innermost circle of the spiral structure, a liquid outlet end 4 of the blood separation channel 1 is positioned at the outermost circle of the spiral structure, a liquid outlet end 4 of the blood separation channel 1 is branched into a centrifugal outer channel and a centripetal inner channel, and the outer channel is connected with the detection channel 2; the detection flow channel 2 comprises a main flow channel, a first branch flow channel 7 and a second branch flow channel 8 which are used for injecting different detection reagents respectively, and the first branch flow channel 7 and the second branch flow channel 8 are communicated with one end of the main flow channel, which is close to the liquid outlet of the blood diversion flow channel 1 respectively; the main flow channel comprises a buffer flow channel 5 and a direct flow channel 6 for detection of the fluorescence measuring instrument, the outer side flow channel, the first branch flow channel 7 and the second branch flow channel 8 converge to an inlet of the buffer flow channel 5, and an outlet of the buffer flow channel 5 is connected with one end of the direct flow channel 6; the buffer flow passage 5 is of a snake-shaped structure; the first branch flow channel 7 is generally filled with FXa reagent, the second branch flow channel 8 is generally filled with fluorescent reagent, and the buffer flow channel 5 is used for enabling the flow to be longer in a certain space so as to enable the FXa and the fluorescent reagent to be fully mixed with blood plasma.

In this embodiment, the micro flow channel of the blood separating channel 1 is designed to be a spiral structure, the liquid inlet end 3 is an opening of the innermost circle of the spiral, and a blood sample enters from the liquid inlet end 3 and gradually flows to the outer circle along the flow channel. The liquid outlet end 4 of the blood separating channel 1 is positioned at the outermost circle and is divided into two channels at the tail. The outer runner which centrifugally deflects towards the outside of the spiral structure is connected with the detection runner 2, and the inner runner which centrifugally deflects towards the inside of the spiral structure is connected with the waste liquid port 9. When blood flows in the spiral flow channel, cells in the blood can generate relative displacement under the action of centrifugal force and gradually enrich towards the inner side of the flow channel, when the blood reaches the bifurcation of the liquid outlet end 4, the blood cells and the plasma respectively enter different branches, the plasma flows to the detection flow channel 2 through the outer side flow channel to complete subsequent measurement, and the blood cells flow to the waste liquid port 9 through the inner side flow channel to be discharged, so that the separation of the whole blood is realized.

The detection flow channel 2 receives the blood plasma conveyed by the blood diversion flow channel 1, and is also provided with two branch flow channels, and the FXa and the fluorescent reagent respectively enter the two branch flow channels and are fully mixed with the blood plasma in the buffer flow channel 5. The outlet of the detection flow channel 2 leads to a waste liquid hole, and waste liquid after detection flows out from the hole.

The micro-fluidic chip in the embodiment adopts the design of the blood separating channel 1 with the spiral structure, and when the micro-fluidic chip is used for separating and detecting whole blood, the anti-interference performance of a blood sample to be detected can be improved, and the detection efficiency is improved; and the test needs less blood, only about 2ml of blood is needed for continuous monitoring for 24 hours, and the damage to the patient is small.

Example 2

The embodiment provides a device for real-time detection of whole blood separation and blood coagulation drugs, as shown in fig. 2, comprising a blood collector (not shown in the figure), a power pump 20, an FXa injection pump 50, a fluorescent reagent injection pump 60, a fluorescence measuring instrument 40, a data processing unit (not shown in the figure) and the microfluidic chip 30, wherein one end of the power pump 20 is connected with the blood collector, the other end of the power pump is connected with a liquid inlet end 3 of a blood splitting flow channel 1, a first branch flow channel 7 is connected with the FXa injection pump 50, a second branch flow channel 8 is connected with the fluorescent reagent injection pump 60, an observation area of the fluorescence measuring instrument 40 faces to a collimation flow channel 6, and the data processing unit is connected with the fluorescence measuring instrument 40; the blood collection device is typically a venous cannula. The fluorescence detection excitation wavelength of the fluorescence measuring instrument 40 is 350nm, and the emission wavelength is 450 nm; the power pump 20 is a peristaltic pump and can be other mechanical pumps; the fluorescence measuring instrument 40 is an Axio Imager 2 fluorescence measuring instrument.

In practical use, the blood or the standard blood sample of the test individual 10 is collected through the venous cannula and enters the peristaltic pump, the peristaltic pump continuously pumps the blood into the liquid inlet end 3 of the microfluidic chip 30, the blood to be tested is subjected to plasma and blood cell separation in the blood separation flow channel 1 of the microfluidic chip 30, the blood cells flow out through the inner side flow channel, and the plasma enters the detection flow channel 2 through the outer side flow channel for continuous measurement; the FXa is continuously injected into a first branch flow channel 7 at the head end of the detection flow channel 2 by an FXa injection pump 50, meanwhile, the fluorescent reagent is continuously injected into a second branch flow channel 8 at the head end by a fluorescent reagent injection pump 60, the injected FXa and the fluorescent reagent are mixed with the separated blood plasma to release fluorescence, when the mixture flows through a tail end straight flow channel 6 (namely an observation position), fluorescence measurement is carried out by a fluorescence measuring instrument 40, the fluorescence detection excitation wavelength is 350nm, the emission wavelength is 450nm, the fluorescence detection excitation wavelength is arranged at the tail end of the microfluidic chip 30 and is used for observing the fluorescence intensity value at the position of the straight flow channel 6, and the fluorescence signal value is recorded in sequence at intervals; and the data processing unit is connected with the fluorescence measuring instrument 40 and obtains the concentration of the FXa inhibitor through a fluorescence measuring value, and specifically comprises the following steps: and (3) calculating the concentration of the FXa inhibitor in the blood sample according to a standard curve method, and drawing a blood concentration-time change curve of the experimental animal after the FXa inhibitor is injected, so as to determine the concentration change condition of the FXa inhibitor in the animal body.

In the embodiment, an anti-FXa method is adopted to specifically measure the FXa inhibitor in blood, so that the blood concentration is directly reflected, and the accuracy is improved; the time for measuring the concentration of the FXa inhibitor is shortened to be within 30 minutes, so that the timeliness is improved; in addition, the fluorescence quantitative method is used in the embodiment, so that the detection sensitivity is improved; the concentration of the FXa inhibitor within the range of 0-1.5IU/ml can be reliably measured, and the measurable range of the concentration is widened; the blood quantity required by the test is small, only about 2ml of blood is needed for continuously monitoring 24 hours, and the injury to the patient is small; the embodiment also optimizes the sampling link, adopts a mechanical pump to automatically sample, reduces manual operation, can obtain concentration measurement data once every 15s, and realizes online real-time monitoring.

It is obvious that the above embodiments of the present invention are only examples for clearly illustrating the present invention, and are not limitations to the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. The utility model provides a micro-fluidic chip for whole blood separation and detection, its characterized in that, including dividing blood runner (1) and detection runner (2), divide blood runner (1) to be helical structure, the feed liquor end (3) that divide blood runner (1) are located helical structure's inner circle the most, divide the play liquid end (4) of blood runner (1) to be located helical structure's outermost circle, divide play liquid end (4) fork of blood runner (1) to be centrifugal outside runner (11) and be used for connecting entad inboard runner (12) of useless liquid mouth, outside runner (11) are connected detection runner (2).

2. The microfluidic chip according to claim 1, wherein the detection flow channel (2) comprises a main flow channel and a first branch flow channel (7) and a second branch flow channel (8) for injecting different detection reagents, respectively, and the first branch flow channel (7) and the second branch flow channel (8) are respectively communicated with one end of the main flow channel close to the outer side flow channel (11).

3. The microfluidic chip according to claim 2, wherein the main channel comprises a buffer channel (5) and a direct current channel (6) for detection of a fluorescence measuring instrument, the outer side channel (11), the first branch channel (7) and the second branch channel (8) converge to an inlet of the buffer channel (5), and an outlet of the buffer channel (5) is connected to one end of the direct current channel (6).

4. The microfluidic chip according to claim 3, wherein the buffer flow channel (5) has a serpentine structure.

5. The utility model provides a whole blood separation and coagulation medicine real-time detection device, its characterized in that, including blood collector, power pump (20), FXa syringe pump (50), fluorescence reagent syringe pump (60), fluorescence measuring apparatu (40), data processing unit and claim 3 micro-fluidic chip (30), power pump (20) one end is connected blood collector, the other end are connected divide the inlet end (3) of blood flow channel (1), first subchannel (7) are connected FXa syringe pump (50), second subchannel (8) are connected fluorescence reagent syringe pump (60), fluorescence measuring apparatu (40) are located the end of detecting flow channel (2), the data processing unit with fluorescence measuring apparatu (40) are connected.

6. The device for real-time detection of whole blood separation and coagulation drugs according to claim 5, wherein the observation area of the fluorescence measuring instrument (40) is aligned with the straight flow channel (6).

7. The device for separating whole blood and detecting coagulation drugs in real time according to claim 5, wherein the blood collector is a venous cannula.

8. The device for real-time detection of whole blood separation and coagulation drugs according to claim 5, wherein the fluorescence detection excitation wavelength of the fluorescence measuring instrument (40) is 350nm, and the emission wavelength is 450 nm.

9. The device for the real-time detection of whole blood separation and coagulation drugs according to claim 5, wherein the power pump (20) is a peristaltic pump.

10. The device for real-time detection of whole blood separation and coagulation drugs according to claim 5, wherein the fluorescence measuring instrument (40) is an Axio Imager 2 fluorescence measuring instrument.