Biochip sampling quantitative correction method and biochip

Abstract

The invention discloses a biological chip sampling quantitative correction method and a biological chip. A blood filtering stop recognition line, a quantitative recognition line and a quantitative start recognition line are sequentially arranged on a quantitative tube of the biological chip, the blood filtering stop recognition line is arranged close to a quantitative inlet, the quantitative start recognition line is arranged close to a quantitative outlet, and the quantitative recognition line is located between the blood filtering stop recognition line and the quantitative start recognition line. When the lower liquid level of the sample reaches the blood filtering stop identification line, stopping the blood filtering action; when the lower liquid level of the sample reaches a quantitative starting identification line, pre-pressing the sample to enable liquid drops to grow at a quantitative outlet, then adding the sample for detection according to a unified quantitative volume or other quantitative volumes according to the relationship between the upper liquid level of the sample and the quantitative identification line, if adding the sample for detection according to the other quantitative volumes, multiplying a detection result by a corresponding coefficient, the corresponding coefficient is equal to the ratio of the unified quantitative volume to the remaining quantitative volumes. The quantitative detection success rate of samples with the same volume and different hematocrites reaches 100% by adopting the method.

Description

Technical Field

[0001] The present invention relates to a biochip, in particular to a biochip sample injection quantitative calibration method and a biochip. Background Art

[0002] In the IVD (in vitro diagnostic products) industry, most detections are performed using plasma / serum. The samples collected in hospitals are whole blood. Since the hematocrit of each sample is different, if the same volume of plasma sample is required, the amount of whole blood needed will vary greatly. The sample volumes of nearly 90% of people are usually normal. For the other 10% of people, in order to obtain the same volume of plasma, the amount of whole blood to be collected will increase significantly. However, when collecting blood, it is impossible to know the hematocrit of the patient's whole blood. Currently, the common practice is to calculate the whole blood collection volume according to the high hematocrit, and then use a biochip for detection.

[0003] As described in Chinese Patent CN109238777A, a filter blood vessel and a quantitative tube are arranged on the biochip. The filter blood vessel is equipped with a filter blood plunger, the quantitative tube is equipped with a quantitative plunger, and the filter blood outlet of the filter blood vessel is communicated with the quantitative inlet at the upper part of the quantitative tube. A reactor is arranged below the quantitative outlet at the lower part of the quantitative tube. When samples with the same volume but different hematocrits are filtered into the quantitative tube through the filter blood vessel, due to the inconsistent sample volumes obtained, it is easy to cause a system error due to insufficient samples when using a unified quantitative volume for detection.

[0004] The biochip is small in volume, and in order to realize the application of fingertip blood, the amount of sample added to the biochip is relatively small. This determines that during the design, the amount of sample filtered into the quantitative tube is not much redundant compared to the amount that needs to be quantitatively added. For example, if the volume to be quantitatively added is 10 μL, the maximum theoretical volume of the sample available for quantification in the quantitative tube is 12 μL. In addition, there are differences in the viscosities of different samples. Due to the viscosity differences, the liquid level positions formed in the quantitative tube will be different, that is, it will affect the volume of the sample filtered into the quantitative tube, resulting in a sample volume available for quantification less than 10 μL. Summary of the Invention

[0005] The technical problem to be solved by the present invention is that for samples with the same volume but different hematocrits, after being filtered into the quantitative tube, the sample volumes are different, resulting in an easy error due to insufficient samples when using a unified quantitative volume for detection. The present invention provides a biochip sample injection quantitative calibration method and a biochip that can quantify and detect samples with the same volume but different hematocrits.

[0006] To solve the above technical problems, the present invention adopts the following technical solutions:

[0007] A method for quantitative calibration of sample injection in a biochip. A blood filter tube and a quantitative tube are arranged on the biochip. The quantitative tube includes a quantitative inlet and a quantitative outlet. The blood filtering outlet of the blood filter tube is communicated with the quantitative inlet of the quantitative tube, where:

[0008] On the quantitative tube, a blood filtering stop identification line, a first quantitative identification line, and a quantitative start identification line are sequentially arranged along the axial direction. The blood filtering stop identification line is arranged close to the quantitative inlet. The quantitative start identification line is arranged close to the quantitative outlet. The first quantitative identification line is located between the blood filtering stop identification line and the quantitative start identification line. The sample volume indicated by the first quantitative identification line is consistent with the unified quantitative volume.

[0009] When the lower liquid level of the sample filtered into the quantitative tube reaches the position of the blood filtering stop identification line, the blood filtering action of the blood filter tube is stopped, and the quantitative action of the quantitative tube is started.

[0010] When the lower liquid level of the sample in the quantitative tube reaches the quantitative start identification line, the sample in the quantitative tube is pressed down by a fixed volume, so that a droplet with a pre-pressured volume grows at the quantitative outlet of the quantitative tube. The pre-pressured volume is less than one-half of the unified quantitative volume.

[0011] After a droplet with a pre-pressured volume is generated at the quantitative outlet of the quantitative tube, if the upper liquid level of the sample in the quantitative tube exceeds or is parallel to the first quantitative identification line, detection is performed by adding samples according to the unified quantitative volume; if the upper liquid level of the sample in the quantitative tube is below the first quantitative identification line, detection is performed by adding samples according to the first quantitative volume, and then the detection result is multiplied by a first coefficient.

[0012] The first quantitative volume is less than the unified quantitative volume, and the first coefficient is equal to the ratio of the unified quantitative volume to the first quantitative volume.

[0013] To further improve the success rate of sample addition, a second quantitative identification line is also arranged on the quantitative tube. The second quantitative identification line is located between the first quantitative identification line and the quantitative start identification line. The sample volume indicated by the second quantitative identification line is less than the unified quantitative volume and greater than one-half of the unified quantitative volume;

[0014] After a droplet with a pre-pressured volume is generated at the quantitative outlet of the quantitative tube, if the upper liquid level of the sample in the quantitative tube is between the first quantitative identification line and the second quantitative identification line, detection is performed by adding samples according to the second quantitative volume, and then the detection result is multiplied by a second coefficient; if the upper liquid level of the sample in the quantitative tube is below the second quantitative identification line, detection is performed by adding samples according to the third quantitative volume, and then the detection result is multiplied by a third coefficient;

[0015] The second quantitative volume is smaller than the first quantitative volume, the second coefficient is equal to the ratio of the uniform quantitative volume to the second quantitative volume, and the third coefficient is equal to the ratio of the uniform quantitative volume to the third quantitative volume.

[0016] Based on the same inventive concept, the present invention also provides a biochip, which includes a blood filtration tube and a quantitative tube, the quantitative tube includes a quantitative inlet and a quantitative outlet, the blood filtration outlet of the blood filtration tube is connected to the quantitative inlet of the quantitative tube, and the blood filtration stop identification line, at least one quantitative identification line (such as the aforementioned first quantitative identification line, second quantitative identification line) and a quantitative start identification line are marked in sequence along the axial direction on the quantitative tube, the blood filtration stop identification line is marked close to the quantitative inlet, the quantitative start identification line is marked close to the quantitative outlet, and the quantitative identification line is located between the blood filtration stop identification line and the quantitative start identification line; the biochip adopts the biochip sampling quantitative correction method and biochip.

[0017] It should be noted that the blood filtration stop identification line, the first quantitative identification line, the second quantitative identification line and the quantitative start identification line in the present invention can be directly marked on the quantitative tube, or can be generated in real time by a visual algorithm system (existing technology) during use.

[0018] In short, the visual algorithm system is a camera connected to a control system with an image processing function set at the position of a biochip in a specific detection instrument, which is used to read a quantitative tube or an image containing a quantitative tube, and obtain the height of the liquid level in the quantitative tube through an image processing program, and compare it with the position of the blood filtration stop identification line, the Nth quantitative identification line and the quantitative start identification line set by the control system, and judge the relationship between the current liquid level and the blood filtration stop identification line, the first quantitative identification line, the second quantitative identification line or the quantitative start identification line, so as to automatically complete the recording of the injection volume and the reading of the corresponding coefficient.

[0019] When the samples after quantification in the present invention are injected in three grades, first, the whole blood volume is taken according to the normal hematocrit. After the sample filtration is completed, since the whole quantification tube cannot be seen, the sample volume in the quantification tube is unknown. At this time, the sample in the quantification tube will be pre-pressed by a set stroke (converted according to the set pre-pressed volume), and droplets with the pre-pressed volume will be formed. After the pre-pressing is completed, the liquid level of the visible part of the quantification tube is identified. When the liquid level exceeds or is parallel to the first quantification identification line (such as the 10 μL identification line), the sample is injected and detected according to the unified quantification volume (such as 10 μL); if the liquid level is between the second quantification identification line (such as the 7 μL identification line) and the first quantification identification line or parallel to the second quantification identification line, the sample is injected and detected according to the second quantification volume (such as 7 μL), and then the detection result is multiplied by the second coefficient = unified quantification volume / second quantification volume; if the liquid level is below the second quantification identification line, the sample is injected and detected according to the third quantification volume (such as 5 μL), and then the detection result is multiplied by the third coefficient = unified quantification volume / third quantification volume. In this way, the detection of samples with a volume less than the unified quantification volume can be realized, avoiding the instrument from reporting an error due to insufficient samples and unable to perform detection.

[0020] The principle of the present invention is as follows: for high- and medium-value samples, there is a linear relationship between the concentration of the analyte in the sample and the luminescence value. Multiplying by a coefficient will not affect the detection result, and even can expand the linear range; for low-value samples, since the detection limit is 10 times or more of the sensitivity, false negatives / false positives will not occur.

[0021] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0022] 1. After adopting the sample injection quantification correction method and the biochip of the present invention, the success rate of quantitative detection of samples with the same volume but different hematocrits reaches 100%. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the following drawings are some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0024] Figure 1 It is a front view of the biochip used in the present invention;

[0025] Figure 2 It is a top view of the biochip used in the present invention;

[0026] Figure 3 It is a schematic diagram of the quantification start identification line of the present invention;

[0027] Figure 4 It is a schematic diagram of the first quantitative identification line, the second quantitative identification line and the pre-pressurized volume;

[0028] Figure 5 It is the quantitative flow chart of the biochip sample injection quantitative calibration method and the biochip of the present invention. Specific embodiments

[0029] The present invention will be further described below in conjunction with specific preferred embodiments, but the protection scope of the present invention is not limited thereby.

[0030] In the description of the present invention, it should be noted that the orientation or positional relationship indicated by the terms "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. is based on the orientation or positional relationship shown in the drawings. It is only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus cannot be construed as a limitation of the present invention. In addition, the terms "first", "second", and "third" are only used for descriptive purposes and cannot be construed as indicating or implying relative importance.

[0031] In the description of the present invention, it should be noted that unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection; it can be directly connected, or indirectly connected through an intermediate medium, and it can be the communication inside two elements. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood in specific situations.

[0032] Please refer to Figure 1 - Figure 4 In an embodiment of the biochip sample injection quantitative calibration method and the biochip of the present invention, it includes a biochip 9, on which a blood filter tube 1 and a quantitative tube 2 are arranged. The quantitative tube 2 includes a quantitative inlet and a quantitative outlet. The blood filtering outlet of the blood filter tube 1 is communicated with the quantitative inlet of the quantitative tube 2, and a reactor 10 is arranged below the quantitative outlet 3 of the quantitative tube 2.

[0033] On the quantitative tube 2, a blood filtration stop identification line 4, a first quantitative identification line 8, a second quantitative identification line 7, and a quantitative start identification line 5 are sequentially arranged along the axial direction. The blood filtration stop identification line 4 is arranged close to the quantitative inlet, the quantitative start identification line 5 is arranged close to the quantitative outlet, the first quantitative identification line 8 and the second quantitative identification line 7 are located between the blood filtration stop identification line 4 and the quantitative start identification line 5, and the sample volume indicated by the first quantitative identification line 8 is consistent with the unified quantitative volume. The sample volume indicated by the second quantitative identification line 7 is less than the unified quantitative volume and greater than one-half of the unified quantitative volume.

[0034] As Figure 5 shown, the specific implementation steps of the biological chip sample injection quantitative calibration method and the first embodiment of the biological chip of the present invention are as follows: Before the start of blood filtration quantification, a blood filtration stop identification line 4 is generated on the quantitative tube 2 by a visual algorithm system (prior art). Then, the blood filtration tube 1 starts the blood filtration action. After the lower liquid level of the sample filtered into the quantitative tube 2 reaches the position of the blood filtration identification line 4, the visual algorithm system issues an order to stop the blood filtration action of the blood filtration tube 1. Then, the quantification action of the quantitative tube 2 starts:

[0035] Before the start of the quantification action, the visual algorithm system 9 generates a quantitative start identification line 5 near the quantitative outlet 3;

[0036] The visual algorithm system controls the quantitative tube 2 to start moving. When the lower liquid level of the sample in the quantitative tube 2 reaches the quantitative start identification line 5, the sample in the quantitative tube 5 is pressed down by a fixed volume using a plunger, so that a droplet with a pre-pressurized volume 6 is generated at the quantitative outlet of the quantitative tube 5. The pre-pressurized volume 6 is less than one-half of the unified quantitative volume;

[0037] At the same time, the visual algorithm system generates a first quantitative identification line 8 and a second quantitative identification line 7 on the quantitative tube 2. After a droplet with a pre-pressurized volume 6 is generated at the quantitative outlet of the quantitative tube 2, if the upper liquid level of the sample in the quantitative tube 2 exceeds or is parallel to the first quantitative identification line 8, the sample is added for detection according to the unified quantitative volume; if the upper liquid level of the sample in the quantitative tube 2 is between the first quantitative identification line 8 and the second quantitative identification line 7 or parallel to the second quantitative identification line 7, the sample is added for detection according to the second quantitative volume, and then the detection result is multiplied by a second coefficient; if the upper liquid level of the sample in the quantitative tube 2 is below the second quantitative identification line, the sample is added for detection according to the third quantitative volume, and then the detection result is multiplied by a third coefficient;

[0038] The sample volume indicated by the second quantitative identification line 7 is less than the unified quantitative volume and greater than one-half of the unified quantitative volume; the second coefficient is equal to the ratio of the unified quantitative volume to the second quantitative volume, and the third coefficient is equal to the ratio of the unified quantitative volume to the third quantitative volume.

[0039] In the method of the present invention, when the analytical instrument calculates the result, the calibration curve is still calculated according to the target value of the unified quantitative volume (such as 10 μL). When quantifying other volumes, it is only necessary to multiply by the second coefficient or the third coefficient.

[0040] In this embodiment, the unified quantitative volume is 10 μL, the second quantitative volume is 7 μL, the third quantitative volume is 5 μL, and the pre-pressurized volume 6 is 4-5 μL. After generating the droplet of the pre-pressurized volume 6, if the upper liquid level of the sample in the metering tube 2 is above the first quantitative recognition line 8 or parallel to the first quantitative recognition line, the quantitative sampling of the unified quantitative volume of 10 μL is controlled to be realized; if the upper liquid level of the sample in the metering tube 2 is between the second quantitative recognition line 7 and the first quantitative recognition line 8 or parallel to the second quantitative recognition line, the quantitative sampling of the second quantitative volume of 7 μL is realized by control, and then the detection result is multiplied by the second coefficient = 10 / 7; if the upper liquid level of the sample in the metering tube 2 is below the second quantitative recognition line 7, but the droplet of the pre-pressurized volume 6 has been generated and there is no bubble, the quantitative sampling of the third quantitative volume of 5 μL is controlled to be realized, and then the detection result is multiplied by the third coefficient = 10 / 5.

[0041] It should be noted that the blood filtration stop recognition line, the first quantitative recognition line, the second quantitative recognition line and the quantitative start recognition line in the present invention can also be directly marked on the metering tube.

[0042] Obviously, in the sample introduction and quantitative calibration method of the biochip and the biochip of the present invention, only the first quantitative recognition line 8 can also be provided on the metering tube 2, that is, the sample is divided into two grades for detection. After the droplet with the pre-pressurized volume 6 is generated at the quantitative outlet of the metering tube 2, if the upper liquid level of the sample in the metering tube 2 exceeds or is parallel to the first quantitative recognition line 8, the detection is carried out according to the unified quantitative volume for sampling; if the upper liquid level of the sample in the metering tube 2 is below the first quantitative recognition line, the detection is carried out according to the first quantitative volume for sampling, and then the detection result is multiplied by the first coefficient; the first quantitative volume is less than the unified quantitative volume, and the first coefficient is equal to the ratio of the unified quantitative volume to the first quantitative volume.

[0043] As described above, it is only the specific implementation scheme of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art can make many possible changes and modifications to the technical solution of the present invention by using the above-disclosed technical content without departing from the scope of the technical solution of the present invention, or modify it into an equivalent embodiment with equivalent changes. Therefore, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention shall fall within the protection scope of the technical solution of the present invention.

Claims

1. A method for quantitative calibration of sample injection in a biochip, wherein a blood filtration tube and a metering tube are arranged on the biochip, the metering tube includes a metering inlet and a metering outlet, and the blood filtration outlet of the blood filtration tube is communicated with the metering inlet of the metering tube, and is characterized in that: A blood filtration stop identification line, a first metering identification line and a metering start identification line are sequentially arranged on the metering tube along the axial direction, the blood filtration stop identification line is arranged close to the metering inlet, the metering start identification line is arranged close to the metering outlet, the first metering identification line is located between the blood filtration stop identification line and the metering start identification line, and the sample volume indicated by the first metering identification line is consistent with the unified metering volume; After the lower liquid level of the sample filtered into the metering tube reaches the position of the blood filtration stop identification line, the blood filtration action of the blood filtration tube is stopped, and the metering action of the metering tube is started; When the lower liquid level of the sample in the metering tube reaches the metering start identification line, the sample in the metering tube is pressed down by a fixed volume, so that a droplet with a pre-pressured volume grows at the metering outlet of the metering tube, and the pre-pressured volume is less than one-half of the unified metering volume; After a droplet with a pre-pressured volume is generated at the metering outlet of the metering tube, if the upper liquid level of the sample in the metering tube exceeds or is parallel to the first metering identification line, detection is performed by adding samples according to the unified metering volume. If the upper liquid level of the sample in the metering tube is below the first metering identification line, detection is performed by adding samples according to the first metering volume, and then the detection result is multiplied by the first coefficient; The first metering volume is less than the unified metering volume, and the first coefficient is equal to the ratio of the unified metering volume to the first metering volume.

2. The biochip sample introduction quantitative calibration method according to claim 1, characterized in that A second metering identification line is further arranged on the metering tube, the second metering identification line is located between the first metering identification line and the metering start identification line, and the sample volume indicated by the second metering identification line is less than the unified metering volume and greater than one-half of the unified metering volume; After a droplet with a pre-pressured volume is generated at the metering outlet of the metering tube, if the upper liquid level of the sample in the metering tube is between the first metering identification line and the second metering identification line, detection is performed by adding samples according to the second metering volume, and then the detection result is multiplied by the second coefficient; if the upper liquid level of the sample in the metering tube is below the second metering identification line, detection is performed by adding samples according to the third metering volume, and then the detection result is multiplied by the third coefficient; The second coefficient is equal to the ratio of the unified metering volume to the second metering volume, and the third coefficient is equal to the ratio of the unified metering volume to the third metering volume.

3. A biochip, comprising a blood filtration tube and a metering tube, the metering tube includes a metering inlet and a metering outlet, and the blood filtration outlet of the blood filtration tube is communicated with the metering inlet of the metering tube, and is characterized in that: A blood filtration stop identification line, at least one metering identification line and a metering start identification line are sequentially marked on the metering tube along the axial direction, the blood filtration stop identification line is marked close to the metering inlet, the metering start identification line is marked close to the metering outlet, and the metering identification line is located between the blood filtration stop identification line and the metering start identification line; The biochip adopts the method for quantitative calibration of sample injection in the biochip according to any one of claims 1-2.

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