A centrifugal microfluidic chip and sample processing method

Abstract

The present invention relates to a centrifugal microfluidic chip and a sample processing method. The centrifugal microfluidic chip comprises a liquid inlet, an aliquot hole, a first siphon pipe, a sample pretreatment area, a second siphon pipe, a reaction hole and a waste liquid pool connected in sequence, and the sample pretreatment area is also connected to an exhaust hole. The aliquot hole comprises two chambers of equal volume to realize equal amount equal division of the sample; the volume of each reaction hole is equal to realize secondary equal division of the sample; the sample pretreatment area and each reaction hole are loaded with different reaction reagents as needed to realize multiple detection applications, or loaded with the same reaction reagent to realize parallel detection of the same sample. The present invention can realize sample pretreatment and multiple aliquot treatments, is convenient to operate, and saves time.

Description

Technical Field

[0001] The invention belongs to the technical field of medical consumables, and in particular relates to a centrifugal microfluidic chip and a sample processing method using the centrifugal microfluidic chip, which can realize sample preprocessing and multiple equal portion processing. Background Art

[0002] Centrifugal microfluidic chips are medical consumables used in the field of instant diagnosis, and can output reaction results quickly and conveniently. Its characteristic is that the microfluidic structure is integrated into a disc-shaped chip, and the flow of microfluids is driven by centrifugal force, thereby realizing sample detection and analysis. The centrifugal system can complete operations such as sample pretreatment, mixing, separation and detection of precise volumes. In recent years, centrifugal microfluidic systems have achieved rapid development with their advantages of high throughput, high integration, multiple parallel analysis, portability, low cost, automation, and low sample and reagent consumption. They have been widely used in biochemical testing, immunoassay, nucleic acid amplification, environmental monitoring, cell sorting, and food safety.

[0003] Patent CN205379906U discloses a multi-purpose microfluidic chip. By limiting the ratio of the main channel cross-sectional area, the sample is distributed more evenly. However, the channel cross-sectional design of the chip with narrow peaks and wide troughs affects the flow rate of the liquid in the main channel, and cannot completely solve the problem of uniform sample distribution and the problem of bubble generation in the main channel.

[0004] Patent 108246373A discloses a centrifugal microfluidic chip, which is based on the U-tube liquid seal principle and is designed with equal cross-sections of the peaks and valleys of the main channel to make the liquid flow rate in the main channel uniform, thereby achieving more uniform distribution of samples. The chip converts unmeasured samples into multiple equal portions of accurate samples through capillary action and centrifugal force when the detection chip rotates, and can achieve multi-index detection, but the above structure is not suitable when the sample needs to be pre-processed. Summary of the invention

[0005] In view of the above problems, the present invention provides a centrifugal microfluidic chip and a sample processing method using the centrifugal microfluidic chip, which can realize sample pretreatment and multiple equal portion processing, is easy to operate and saves time.

[0006] The technical solution adopted by the present invention is as follows:

[0007] A centrifugal microfluidic chip comprises a liquid inlet, an equal portion hole, a first siphon pipe, a sample pretreatment area, a second siphon pipe, a reaction hole and a waste liquid pool which are connected in sequence, wherein the sample pretreatment area is also connected to an exhaust hole.

[0008] Furthermore, the equal-division hole includes two chambers of equal volume to achieve equal division of the sample.

[0009] Furthermore, there are at least two reaction holes.

[0010] Furthermore, the volume of each reaction well is equal to achieve secondary equal division of the sample.

[0011] Furthermore, the sample pretreatment area and each reaction well are loaded with different reaction reagents as required to realize multiple detection applications; or loaded with the same reaction reagents to realize parallel detection of the same sample.

[0012] Furthermore, the reagent is loaded in one of the following ways: freeze drying, vacuum drying, and high temperature drying.

[0013] Furthermore, it also includes a cover plate or a cover film, which is used to form a complete structure by sealing; the sealing method is one of the following: ultrasonic bonding, hot pressing bonding, and simple film pasting treatment.

[0014] A centrifugal microfluidic system comprises the centrifugal microfluidic chip described above and a centrifuge; the centrifugal microfluidic chip is mounted on the centrifuge, and the centrifuge drives the microfluid in the centrifugal microfluidic chip to flow by centrifugal force.

[0015] A sample processing method using the centrifugal microfluidic chip comprises the following steps:

[0016] 1) Add a certain amount of sample into the liquid inlet;

[0017] 2) starting the centrifuge, and after a certain period of time at the first speed, the sample is evenly divided into equal amounts at the equal-dividing holes;

[0018] 3) standing for a certain period of time to allow the sample to fill the first siphon channel through capillary action;

[0019] 4) The centrifuge is started for the second time, and after a certain period of time at the second speed, the sample enters the sample pretreatment area;

[0020] 5) Let it stand for a certain period of time to allow the sample to fill the second siphon channel through capillary action;

[0021] 6) The centrifuge is started for the third time. After a certain period of time at the third speed, the samples enter the reaction wells and the waste liquid pool in turn.

[0022] Furthermore, in step 2), the first rotation speed is 100-3000rpm / s, and the certain time is 20-40s; in step 4), the second rotation speed is 100-3000rpm / s, and the certain time is 30-100s; in step 6), the third rotation speed is 100-3000rpm / s, and the certain time is 30-100s.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] 1) The centrifugal microfluidic chip designed in the present invention can not only realize the bisection of samples, but also pre-treat and perform secondary aliquoting on the aliquoted samples, which is suitable for the situation where the samples need to be pre-treated and aliquoted multiple times.

[0025] 2) The centrifugal microfluidic chip designed in the present invention only requires one step of sample addition operation for operators, thus avoiding the tedious sample pretreatment process, requiring no professional training, being easy to operate, and saving time. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Schematic diagram of the structure of the centrifugal microfluidic chip in Example 1.

[0027] Figure 2 Schematic diagram of the structure of the centrifugal microfluidic chip in Example 2. DETAILED DESCRIPTION

[0028] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the present invention is further described in detail below through specific embodiments and drawings.

[0029] Embodiment 1:

[0030] Figure 1 1 is a schematic diagram of the structure of the centrifugal microfluidic chip 100 of this embodiment. The centrifugal microfluidic chip 100 includes a liquid inlet 101, an aliquot hole 102, a first siphon pipe 103, a sample pretreatment area 104, an exhaust hole 105, a second siphon pipe 106, reaction holes 107 and 108, and a waste liquid pool 109. Figure 1 There are two groups 101~109, and these two groups 101~109 are connected by 102.

[0031] The centrifugal microfluidic chip 100 is installed on a centrifuge to form a centrifugal microfluidic system to process samples, including the following steps:

[0032] 1) Add a certain amount of sample into the liquid inlet 101.

[0033] 2) Start the centrifuge at a speed within the range of 100-3000 rpm / s and a time within the range of 20-40 seconds. Since the aliquot hole 102 includes two aliquot structures, i.e., two chambers of equal volume, the liquid surface after centrifugation is at the same arc, so the sample can be equally divided at the aliquot hole 102.

[0034] 3) After standing for 10-60 seconds, the sample fills the first siphon channel 103 through capillary action.

[0035] 4) The centrifuge is started for the second time, with the speed in the range of 100-3000 rpm / s and the time in the range of 30-100 s, and the sample enters the sample pretreatment area 104 .

[0036] 5) After standing for 100-300 seconds, the solution fills the second siphon pipe 106 by capillary action.

[0037] 6) The centrifuge is started for the third time, with a rotation speed in the range of 100-3000 rpm / s and a time in the range of 30-100 s, and the solution sequentially enters the reaction wells 107, 108, and the waste liquid pool 109. The volumes of the reaction wells 107 and 108 can be set equal, so that the solution is divided twice at the reaction wells 107 and 108.

[0038] Among them, 100 is used as a chip substrate and can be sealed with a cover plate or a cover film to form a complete structure. The sealing method can be ultrasonic bonding, hot pressing bonding, or simple film bonding.

[0039] The sample pretreatment area 104 and the reaction wells 107 and 108 can be loaded with different reaction reagents as needed to realize multiple detection applications, or loaded with the same reaction reagents to realize parallel detection of the same sample. The reagent loading method can be freeze drying, vacuum drying, high temperature drying, etc.

[0040] After the centrifuge is started for the third time, the reaction holes 107 and 108 are completely filled, and the excess liquid enters the waste liquid pool 109.

[0041] Embodiment 2:

[0042] Figure 1 In the structure of the embodiment 1 shown, there are two groups 101-109 on the centrifugal microfluidic chip 100. In this embodiment, more groups 101-109 are provided and evenly distributed on the centrifugal microfluidic chip 100, such as Figure 2 The structure shown in the figure improves the sample processing efficiency.

[0043] The other structures of the centrifugal microfluidic chip of this embodiment are the same as those of Embodiment 1.

[0044] Embodiment 3:

[0045] Figure 1 In the structure of the embodiment 1 shown, there are two reaction holes, namely 107 and 108. In this embodiment, one or more than two reaction holes are provided.

[0046] The other structures of the centrifugal microfluidic chip of this embodiment are the same as those of Embodiment 1.

[0047] The above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the same. A person skilled in the art may modify or make equivalent substitutions for the technical solutions of the present invention without departing from the spirit and scope of the present invention. The protection scope of the present invention shall be subject to the claims.

Claims

1. A centrifugal microfluidic chip, characterized in that: It includes a liquid inlet hole, an equal portion hole, a first siphon pipe, a sample pretreatment area, a second siphon pipe, a reaction hole and a waste liquid pool which are connected in sequence, and the sample pretreatment area is also connected to an exhaust hole; the equal portion hole includes two chambers with equal volumes to achieve equal amounts of sample division; there are at least two reaction holes, each of which has an equal volume to achieve secondary equal division of the sample.

2. The centrifugal microfluidic chip according to claim 1, characterized in that: The sample pretreatment area and each reaction well are loaded with different reaction reagents as needed to realize multiple detection applications; or loaded with the same reaction reagents to realize parallel detection of the same sample.

3. The centrifugal microfluidic chip according to claim 2, characterized in that: The reagent is loaded in one of the following ways: freeze drying, vacuum drying, and high temperature drying.

4. The centrifugal microfluidic chip according to claim 1, characterized in that: It also includes a cover plate or a cover film, which is used to form a complete structure through sealing; the sealing method is one of the following: ultrasonic bonding, hot pressing bonding, and simple film pasting treatment.

5. A centrifugal microfluidic system, characterized in that: It comprises the centrifugal microfluidic chip as claimed in any one of claims 1 to 4, and a centrifuge; the centrifugal microfluidic chip is installed on the centrifuge, and the centrifuge drives the microfluid in the centrifugal microfluidic chip to flow through centrifugal force.

6. A sample processing method using the centrifugal microfluidic chip according to claim 1, characterized in that: The following steps are involved: 1) Add a certain amount of sample into the liquid inlet; 2) starting the centrifuge, and after a certain period of time at the first speed, the sample is evenly divided into equal amounts at the equal-dividing holes; 3) standing for a certain period of time to allow the sample to fill the first siphon channel through capillary action; 4) The centrifuge is started for the second time, and after a certain period of time at the second speed, the sample enters the sample pretreatment area; 5) Let it stand for a certain period of time to allow the sample to fill the second siphon channel through capillary action; 6) The centrifuge is started for the third time. After a certain period of time at the third speed, the sample enters the reaction wells and the waste liquid pool in turn. The volume of each reaction well is equal, and the sample is divided twice through the reaction wells.

7. The method according to claim 6, characterized in that Step 2) the first speed is 100-3000 rpm / s, and the certain time is 20-40s; Step 4) the second speed is 100-3000 rpm / s, and the certain time is 30-100s; Step 6) The third rotation speed is 100-3000 rpm / s, and the certain time is 30-100 s.

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