TWI868835B - Microfluidic cassette and detection method - Google Patents

Abstract

Disclosed herein is a microfluidic cassette which has a test cassette and inserting member, wherein the inserting member is configured to insert into the test cassette. The base has a receiving well, a plurality of reaction wells, a plurality of channels and a plurality of indication wells disposed on the base. The upper cover is disposed on the base and has an opening for receiving the fluid sample, a plurality of testing holes respectively corresponding to the reaction wells for analysis, and a fluid guiding member. The inserting member has a plate member and a plurality tubes on the plate member. When the inserting member inserted into the test cassette, the plate member of the inserting member is on the upper cover, and the tubes of the inserting member are respectively inserted into the reaction wells, so as to retain the fluid sample within the tube members of the insert member.

Description

Microfluidic cartridge and detection method

The present invention relates to a microfluidic cartridge, and in particular to a microfluidic cartridge that can complete a detection reaction without the need for external power.

Micro-test reagents can complete various clinical tests with low sample volumes and have many advantages. In development, the micro-test reagents can be used in conjunction with a variety of immunological binding methods (e.g., direct detection, sandwich method, or competitive method) to complete various tests. For medical tests, micro-test reagents are convenient, fast, and safe, and have many advantages.

However, in the detection of the microfluidic channel of the mesenchymal test, it is also necessary to use external power, use a micro pump to drive the sample fluid to flow through the pressure difference, and use a micro valve to control the flow direction of the sample fluid on the test microfluidic channel. It can be seen that the microfluidic cartridge of the previous technology has its limitations in use.

In view of this, in order to improve the defects of the previous technology, this field urgently needs an improved test reagent to improve the shortcomings of the previous technology.

The content of the invention is intended to provide a simplified summary of the disclosure so that readers can have a basic understanding of the disclosure. This content of the invention is not a complete overview of the disclosure, and it is not intended to point out the important/key elements of the embodiments of the invention or to define the scope of the invention.

To solve the defects of the prior art, one aspect of the present invention is a novel microfluidic cartridge that can complete the inspection step without external power. Specifically, the microfluidic cartridge includes a test cartridge and a plug-in. The test box includes a base and an upper cover, wherein the base is provided with a receiving tank, a plurality of reaction tanks, a plurality of drainage channels, a plurality of indicator tanks and a collection tank, wherein the receiving tank includes a plurality of buffer flow channels, a plurality of reaction tanks are located downstream of the receiving tank, and one end of each drainage channel is connected to the receiving tank fluid, and the other end is respectively connected to the reaction tank fluids to guide the sample fluid from the receiving tank to the reaction tank, the indicator tanks are respectively connected to the reaction tank fluids to receive the sample fluids flowing out of the reaction tanks, and the collection tank is connected to the indicator tank fluid to receive the sample fluid flowing out of the indicator tank. The upper cover is arranged on the base, and an injection hole, a plurality of inspection holes and a flow guide are arranged on the structure. The injection hole is arranged corresponding to the receiving groove on the base, and the inspection hole is arranged corresponding to the reaction grooves; the flow guide is located below the upper cover, and is corresponding to the buffer flow channels or the drainage channels of the receiving groove, and is connected with the fluid of the buffer flow channels or the drainage channels to guide the sample fluid from the receiving groove to the reaction groove. The plug-in is used to be inserted into the multiple reaction tanks of the microfluidic cartridge, and the structure includes a plate-shaped member and multiple tubes, wherein the tubes are arranged on the plate-shaped member and are arranged corresponding to the reaction tanks, wherein the tubes are configured to penetrate the inspection holes of the upper cover and are inserted into the reaction tanks of the base, so that the sample fluids in the reaction tanks are retained in the tubes.

In addition, in order to prevent the sample fluid from flowing back, a first tilt angle is configured at the connection between the reaction tanks and the indicator tank fluids in the microfluidic cartridge, and a second tilt angle is configured at the connection between the indicator tank sample fluids and the collection tank to prevent the sample fluid from flowing back into the indicator tank.

According to a specific implementation of the present invention, the reaction tanks and indicator tanks in the microfluidic cartridge of the present invention are three in number and arranged in a row, and the fluid connection between the reaction tank located in the middle of the reaction tanks and the corresponding indicator tank is fan-shaped.

In one embodiment of the present invention, the microfluidic cartridge further includes a sample cover fitted on the injection hole. In other embodiments, it further includes a reaction tank cover for detachably covering the tubular objects of the plug-in.

According to an embodiment of the present invention, the microfluidic cartridge further includes a viewing window, which is arranged corresponding to the indicator slot.

According to an implementation method of the present invention, the microfluidic cartridge of the present invention further includes a plurality of anti-overflow rings respectively disposed on the upper edges of a plurality of inspection holes.

According to a specific implementation of the present invention, the bottom of the base of the microfluidic cartridge corresponding to the plurality of reaction tanks protrudes more than the bottom around the base.

Another embodiment of the present invention is a method for testing using the microfluidic cartridge described in any of the above embodiments, the method comprising the following steps: (a) injecting a sample fluid into the base through the injection hole of the upper cover, wherein the sample fluid flows through the receiving tank through the drainage channels to the reaction tank, the indicator tank and the collection tank respectively; (b) inserting the plug-in through the upper cover into the base, wherein the tubes of the plug-in are respectively inserted into the reaction tanks of the base, so that the sample fluid is retained in the tubes; and (c) detecting the content of a test substance in the sample fluid in the tubes.

In a specific implementation, the step (c) is to detect the fluorescence value of the sample fluid in the tubes through a fluorescence detection device.

After reading the implementation method below, a person with ordinary knowledge in the technical field to which the present invention belongs can easily understand the basic spirit and other invention purposes of the present invention, as well as the technical means and implementation methods adopted by the present invention.

The main component symbols of this invention are listed as follows:

1000, 7000: Microfluidic cartridge

100: Inspection box

104: View window

120: Base

122: receiving slot

123: Buffer channel

124, 124M: reaction tank

126: Drainage channel

128: indicator slot

130: Collection slot

160: Upper cover

162: Injection hole

163: Hiatus

164: Inspection hole

166: Flow guide

167A, 167B: deflector

168, 168a, 168b, 168c: flow channel

170: Anti-overflow ring

180:Plugins

182: Plate-like parts

184: Tube

190: Reactor cover

192: Sample cover

193: Stickers

S: Sample fluid

In order to make the above and other objects, features, advantages and embodiments of the present invention more clearly understood, the attached drawings are described as follows: FIG. 1A and FIG. 1B are schematic diagrams of a microfluidic cartridge 1000 according to an embodiment of the present invention; FIG. 2A is a top view of a base 120 of the microfluidic cartridge shown in FIG. 1A; FIG. 2B is a cross-sectional view of the base 120 drawn along line 2A-2A of FIG. 2A; FIG. 2C is a bottom view of the base 120 of the microfluidic cartridge shown in FIG. 2A; FIG. 3A is a top view of a cover 160 of the microfluidic cartridge shown in FIG. 1A; FIG. 3B is a cross-sectional view of the base 120 drawn along line 2A-2A of FIG. 2A; FIG. 3C is a bottom view of the base 120 of the microfluidic cartridge shown in FIG. 2A; FIG. 3A is a top view of a cover 160 of the microfluidic cartridge shown in FIG. Figure B is a bottom view of the upper cover 160 of the microfluidic cartridge; Figure 3C is a cross-sectional view of the upper cover 160 according to the 3C-3C line of Figure 3B; Figures 4A and 4B are schematic diagrams of the structure of the plug-in 180 shown in Figure 1A; Figure 5 is a cross-sectional view of the microfluidic cartridge 1000 shown in Figure 1A; Figures 6A to 6C are schematic diagrams of the structure of the microfluidic cartridge 7000 shown in another embodiment of the present invention; and Figures 7A to 7E are schematic diagrams of performing an inspection method using the microfluidic cartridge 1000 shown in Figures 1A and 1B.

According to conventional working methods, the various features and components in the figure are not drawn to scale. The drawing method is to present the specific features and components related to the present invention in the best way. In addition, the same or similar component symbols are used to refer to similar components/parts between different figures.

In order to make the description of the disclosure more detailed and complete, the following provides an illustrative description of the implementation and specific embodiments of the present invention; however, this is not the only form of implementing or using the specific embodiments of the present invention. The implementation covers the features of multiple specific embodiments and the method steps and their sequence for constructing and operating these specific embodiments. However, other specific embodiments can also be used to achieve the same or equal functions and step sequences.

Although the numerical ranges and parameters used to define the broader scope of the present invention are approximate, the relevant numerical values in the specific embodiments have been presented as accurately as possible. However, any numerical value inherently inevitably contains standard deviations caused by individual testing methods. Here, "about" generally means that the actual value is within plus or minus 10%, 5%, 1% or 0.5% of a particular value or range. Alternatively, the word "about" means that the actual value falls within the acceptable standard error of the mean, depending on the consideration of a person of ordinary skill in the art to which the present invention belongs. Except for the experimental examples, or unless otherwise expressly stated, it is understood that all ranges, quantities, values and percentages used herein (for example, to describe the amount of material used, the length of time, temperature, operating conditions, quantity ratios and the like) are modified by "about". Therefore, unless otherwise stated to the contrary, the numerical parameters disclosed in this specification and the attached patent application are approximate values and can be changed as needed. At least these numerical parameters should be understood as the indicated significant digits and the values obtained by applying the general rounding method.

The term "sample fluid" herein refers to a sample suitable for use with the microfluidic cartridge 100 of the present invention, preferably a biological sample. In an optional embodiment, the biological sample may be blood, body fluid, saliva or other secretions. In another optional embodiment, the biological sample may be a processed biological sample, for example, a biological sample pre-treated with a buffer solution.

Unless otherwise defined in this specification, the scientific and technical terms used herein have the same meanings as those understood and used by persons of ordinary knowledge in the technical field to which the present invention belongs. In addition, singular terms used in this specification include the plural form of the term, and plural terms also include the singular form of the term, unless otherwise conflicting with the context.

The present invention proposes a novel microfluidic cartridge, which can complete the detection reaction without the assistance of other devices to provide external force (for example, gas pressurization) through the design of the flow channel and various components. Furthermore, the microfluidic cartridge of the present invention is equipped with multiple reaction tanks, which can detect different test items for a single sample at the same time, or detect multiple samples at the same time.

Figures 1A and 1B are schematic diagrams of a microfluidic cartridge 1000 according to an embodiment of the present invention, wherein Figure 1A is an exploded schematic diagram of the microfluidic cartridge 1000. The microfluidic cartridge 1000 of the present invention mainly includes a test cartridge 100 and a plug-in 180.

As shown in FIG. 1A, the test box 100 includes a base 120 and an upper cover 160, which are coupled to each other up and down to form the test box 100. A person with ordinary knowledge in the field of the present invention should understand that the coupling structure of the upper cover 160 and the base 120 can be completed by using the existing snap-fit structure or combination method in the field of the present invention. Next, please refer to FIG. 1B at the same time. The plug-in 180 is detachably inserted into the base 120 and the upper cover 160 of the test box 100.

To clearly understand the structure of the base 120 of the test cassette 100, please refer to Figures 2A to 2C simultaneously, wherein Figure 2A is a top view of the base 120 of the test cassette shown in Figure 1; Figure 2B is a cross-sectional view of the base 120 drawn along the line 2B-2B of Figure 2A; and Figure 2C is a bottom view of the base 120 of the microfluidic cassette.

As shown in FIG. 2A , the base 120 includes a receiving tank 122, a plurality of reaction tanks 124, a plurality of drainage channels 126, a plurality of indicator tanks 128, and a collection tank 130 disposed on the base 120 and fluidically connected to each other. In a specific embodiment, the configuration of the tanks of the base 120 from upstream to downstream is sequentially the receiving tank 122, the reaction tank 124, the indicator tank 128, and the collection tank 130, that is, the sample fluid (e.g., the test sample) enters the reaction tank 124 from the receiving tank 122, and the excess sample fluid will overflow to the indicator tank 128 and the collection tank 130. In a preferred embodiment, in the base 120 of the microfluidic cartridge of the present invention, the flow direction of the sample fluid in the receiving tank 122, the reaction tank 124 and the indicator tank 128 is set to be a single flow direction, that is, the sample fluid does not flow back.

In the present embodiment, the receiving tank 122 is provided with four buffer channels 123 which are arranged radially, wherein one end of the four buffer channels 123 is close to each other and connected to the three drainage channels 126. It should be noted that the buffer channel 123 is in the shape of a water drop, and the diameter of the end not connected to the drainage channel 126 is larger than the diameter of the end connected to the drainage channel 126. The buffer channel 123 is then connected to the three reaction tanks 124 respectively through the drainage channels 126 to guide the sample fluid from the receiving tank 122 to each of the reaction tanks 124 to facilitate the reaction. In this embodiment, three indicator slots 128 are provided, which are respectively connected to the fluid of each reaction slot 124. The sample fluid overflowing from the reaction slot 124 will enter the indicator slot 128 and then flow into the collection slot 130. In addition, the buffer channel 123 on the receiving slot 122 enters the drainage channel 126. The inclination of the buffer channel 123 is at least about 1 degree, preferably 1.5 degrees, to assist the sample fluid to flow into the drainage channel 126 and prevent it from flowing back to the buffer channel 123.

In order to prevent the sample fluid from flowing back, a tilt angle design may be provided between each slot of the microfluidic cartridge to prevent the sample fluid from flowing back. Please refer to Figures 2B and 2C. A first tilt angle A1 is configured at the fluid connection between the reaction slot 124 and the indicator slots 128, and a second tilt angle A2 is configured at the connection between the sample fluid of the indicator slot 128 and the collecting slot 130 to prevent the sample fluid from flowing back into the indicator slot. The first tilt angle is specifically tilted from the reaction slot 124 to the indicator slot 128, that is, the plane corresponding to the fluid connection of the reaction slot 124 is higher than the plane height of the indicator slot 128; on the other hand, the second tilt angle is tilted from the indicator slot 128 to the collecting slot 130, and the plane close to the indicator slot 128 is higher than the plane close to the collecting slot 130. The angles of the first tilt angle and the second tilt angle are about at least 10 degrees, for example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 and 20, preferably at least 15 degrees.

Furthermore, please refer to Figure 2A again. This embodiment has three reaction tanks 124, and the fluid connection between the middle reaction tank 124M and the corresponding indicator tank 128 is fan-shaped. This design is to allow the sample fluid to flow to the indicator tank 128 and the recovery tank 130 at a constant speed.

In addition, please refer to Figure 2C, which is a bottom view of the base 120. According to an embodiment of the present invention, the bottom of the base 120 corresponding to the reaction tank 124 is more protruding than the surrounding area. As shown in the figure, the part corresponding to the reaction tank 124 protrudes out of the bottom of the base 120 to facilitate heating the reaction tank 124. Furthermore, in order to increase the volume of the collection tank 130, the bottom corresponding to the collection tank 130 can also protrude from the bottom of the base 120.

FIG. 3A is a top view of the upper cover 160 shown in FIG. 1, and FIG. 3B is a bottom view of the upper cover 160. Specifically, the upper cover 160 is coupled to the base 120 to form the test box 100, and is preferably connected by a snap-fit structure. The upper cover 160 is provided with an injection hole 162, a plurality of inspection holes 164, and a guide 166 on the structure. The injection hole 162 is provided corresponding to the receiving slot 122 on the base 120, and the plurality of inspection holes 164 are provided corresponding to the reaction slots 124 respectively. The guide 166 is provided corresponding to the buffer channels 123 and/or the drainage channels 126 of the receiving slot 122 and is fluidly connected to guide the sample fluid from the receiving slot 122 to the reaction slot 124.

FIG. 3C is a cross-sectional view of the upper cover 160 to clearly show the structure of the guide member 166. Please refer to FIG. 3C and FIG. 2A at the same time. The guide member 166 is structurally composed of at least two guide plates, namely guide plates 167A and 167B. The guide plates 167A and 167B are provided with an inclined angle on the outer side, and each of the guide plates 167A and 167B defines a flow channel 168 at a distance interval, so that the guide member 166 and the buffer flow channels 123 of the receiving tank 122, the drainage channels 126 and the reaction tanks 124 are fluidly connected.

In addition, three viewing windows 104 may be provided on the upper cover 160, which are respectively provided corresponding to the three indicator slots 128. Through the viewing windows 104, it is possible to check whether the sample fluid flows into the indicator slots 128, which can be used to confirm whether the test sample volume is sufficient. In other embodiments, the plurality of viewing windows 104 may also be a single viewing window, and the opening range of the single viewing window can be provided corresponding to the indicator slots 128, so as to be used to observe whether the sample fluid flows into the indicator slots.

Please refer to Figure 3A again. The injection hole 162 is specially designed. The injection hole 162 is provided with a crack 163, and below it is a portion of the guide 166, which corresponds to the configuration of the buffer flow channel 123. The crack structure design of the crack 163 makes the sample fluid produce a retention effect in the injection hole 162 after the sample is injected into the injection hole 162, and then guided to the reaction tank 124 through the guide 166.

In this embodiment, the base 120 of the microfluidic cartridge 100 of the present invention is provided with three reaction tanks 124. In order to control the flow rate and flow velocity of the sample fluid entering each of the reaction tanks 124, the widths of the flow channels 168a, 168b and 168c corresponding to the drainage channels 126 can be adjusted. For example, the widths of the two outer channels 168a and 168c are greater than the width of 168b, so that the speed of the sample fluid flowing from the receiving tank 122 to each of the reaction tanks 124 is close to the same.

According to other embodiments of the present invention, the microfluidic cartridge 100 of the present invention further includes a plurality of anti-overflow rings 170 respectively disposed on the upper edges of a plurality of inspection holes 164.

It can be seen from this that the microfluidic cartridge 100 proposed by the present invention allows the sample fluid to enter the buffer channel 123 through the injection hole 162 through the guide member 166, and then enter the reaction tank 124 through the drainage channel 126. Excessive samples overflow into the indicator tank 128 and then enter the collection tank 130.

Please refer to FIG. 4A and FIG. 4B for schematic diagrams of the structure of the plug-in 180 shown in FIG. 1. The plug-in 180 structurally includes a plate-like member 182 and a plurality of tubes 184 arranged on the plate-like member 182. Please also refer to FIG. 5, the plurality of tubes 184 on the plug-in 180 are arranged corresponding to the reaction tanks 124 of the base 120, wherein the tubes 184 are configured to penetrate the inspection holes 164 of the upper cover 160, and are inserted into the reaction tanks 124 of the base 120, so that the sample fluids in the reaction tanks 124 are retained in the tubes 184. Furthermore, it should be noted that the present invention can retain the sample fluid in the tubular object 184 through the setting of the plug-in 180, so that the samples in each reaction tank 124 can be kept at the same height, thereby avoiding the loss of fluid samples, reducing contamination, and improving the accuracy of biological sample detection.

Figures 6A to 6C show the microfluidic cartridge 7000 according to various embodiments of the present invention. In these figures, the microfluidic cartridge 7000 includes a reaction tank cover and/or a sample cover for sealing the cartridge. For example, the microfluidic cartridge 7000 before the reaction can be stored in the microfluidic cartridge 7000 through this setting to avoid overflow or contamination.

Please refer to Figure 6A first. The microfluidic cartridge 7000 further includes a reaction tank cover 190 and/or a sample cover 192 that is detachably covered on the inspection hole 164 and the injection hole 162 of the cover 160 (also refer to Figure 3A).

Furthermore, in the implementation of FIG. 6B, the sample cover is replaced by a sticker, and the sticker 193 is used to seal the test hole 164. In actual use, when the microfluidic cartridge 7000 is to be used for testing, the sticker 193 can be torn off first to expose the reaction hole 164, and the microfluidic cartridge 7000 of the present invention can be used for testing.

In addition, in the embodiment of FIG. 6C, the reaction tank cover 190 is designed as a sliding cover. The reaction tank cover 190 can be movably engaged with the test box 100 and moved along the length direction of the test box 100. Those with common knowledge in the technical field to which the present invention belongs should understand that the setting method of the sliding cover can be changed according to actual use requirements and common knowledge in this field.

Another aspect of the present invention is a method for testing using a microfluidic cartridge shown in any embodiment of the present invention. Please refer to Figures 7A to 7E. In this embodiment, the method for testing using the microfluidic cartridge 1000 shown in Figure 1 is as follows. First, as shown in Figure 7A, the sample fluid S to be tested (for example, a diluted or pre-treated liquid test sample) is injected into the test cartridge 100 through the injection hole 162, wherein the sample fluid S passes through the receiving tank 122 through the drainage channel 126 to the reaction tank 124, and then enters the collection tank 130 through the indicator tank 128 (please also refer to Figures 2A and 2B).

In addition, whether the volume of the test sample in the microfluidic cartridge 1000 is sufficient for measurement can also be confirmed through the viewing window 104 to see whether the sample volume in each reaction tank 124 is sufficient. The sample volume can be confirmed by the naked eye or by related instruments.

Next, please refer to Figures 7B to 7D, insert the plug-in 180 into the test box 100, wherein the tube 184 passes through the test holes 164 of the upper cover 160 and is inserted into the reaction tanks 124 of the base 120, so that the sample fluid S in the reaction tanks 124 is retained in the tubes 184 (please also refer to Figure 3). Please refer to Figure 7D, when the plug-in 180 is inserted into the test box 100, it can ensure the loss of the sample fluid S in the test box 100.

For example, during the testing process, instruments are usually required to interpret the test results. In order to prevent the microfluidic cartridge 100 from shaking and causing the sample fluid S to flow out of the reaction tank 124, so that the volume of the test sample in each reaction well is different, thereby affecting the accuracy of the test, the present invention can make the sample fluid S stay in the reaction tank 124 through the setting of the plug-in 180, and the sample fluid S in each reaction tank 124 can maintain a certain volume, and each sample fluid S is maintained at the same height, so as to maintain the stability of biological sample testing.

In this step, the microfluidic cartridge 1000 of the present invention can be tested in combination with existing testing methods in the technical field of the present invention. For example, after inserting the plug-in 180 into the test cartridge 100, the instrument is used to perform a nucleic acid amplification reaction first, and then the enzyme immunoassay, fluorescent immunoassay, nucleic acid fluorescence or colorimetric assay is used for measurement. Enzyme immunoassay or nucleic acid fluorescence or colorimetric assay is used for measurement

In addition, the sample cover 192 can be placed on the injection hole 162 as needed to prevent the test sample from splashing during operation, which may cause cross contamination between samples.

Finally, please refer to Figure 7E. The result after the reaction (e.g., color reaction or fluorescence reaction) can be read through the through hole of the tubular object 184 of the plug 180 (i.e., the inspection hole 164 corresponding to the upper cover 160) with the relevant inspection equipment D, for example, by measuring the absorbance value or fluorescence value.

In one embodiment of the present invention, the reaction result can be detected by a fluorescence detection device to detect the fluorescence value of the sample fluid S in the tubular objects 184.

The microfluidic cartridge proposed by the present invention is suitable for the field of environmental testing or medical testing, especially for the field of medical testing. A person with ordinary knowledge in the relevant technical field can design and coat or externally place the reaction substances (such as nucleic acid probes, antigens, antibodies or other substances) in the reaction tank of the microfluidic cartridge according to actual use requirements to facilitate the detection of the test substances in the sample fluid.

Although the above embodiments disclose specific embodiments of the present invention, they are not intended to limit the present invention. Those with ordinary knowledge in the technical field to which the present invention belongs can make various changes and modifications without departing from the principles and spirit of the present invention. Therefore, the scope of protection of the present invention shall be based on the scope defined in the accompanying patent application.

1000: Microfluidic cartridge

100: Inspection box

120: Base

160: Upper cover

162: Injection hole

164: Inspection hole

180:Plugins

Claims (10)

A microfluidic cartridge for processing a sample fluid comprises: a test cartridge comprising a base and a cover engaging with each other, wherein: the base comprises: a receiving tank comprising a plurality of buffer channels, arranged on the base; a plurality of reaction tanks, arranged on the base and located downstream of the receiving tank; a plurality of drainage channels, one end of which is connected to the receiving tank fluid, and the other end of which is respectively connected to the reaction tank fluids, for guiding the sample fluid from the receiving tank to the reaction tanks; a plurality of indicator tanks, respectively connected to the reaction tank fluids, for receiving the sample fluid flowing out of the reaction tanks; and a collection tank, connected to the indicator tank fluids, for receiving the sample fluid flowing out of the indicator tanks; the cover, arranged on the base, comprises: an injection hole, located downstream of the receiving tank; a plurality of drainage channels, one end of which is connected to the receiving tank fluid, and the other end of which is respectively connected to the reaction tank fluids, for guiding the sample fluid from the receiving tank to the reaction tanks; a plurality of indicator tanks, respectively connected to the reaction tank fluids, for receiving the sample fluid flowing out of the indicator tanks; and a collection tank, connected to the indicator tank fluids, for receiving the sample fluid flowing out of the indicator tanks; the cover, arranged on the base, comprises: an injection hole, located downstream of the receiving tank; a plurality of drainage channels, one end of which is connected to the receiving tank fluid, and the other end of which is respectively connected to the reaction tank fluids, for guiding the sample fluid from the receiving tank to the reaction tanks; a plurality of indicator tanks, respectively connected to the reaction tank fluids, for receiving the sample fluid flowing out of the indicator tanks; and a collection tank, The cover is provided corresponding to the receiving tank; a plurality of inspection holes are located in the cover and respectively provided corresponding to the reaction tanks; and a flow guide is located below the cover and provided corresponding to the buffer flow channels or the drainage channels of the receiving tank and is in fluid communication with the buffer flow channels or the drainage channels to guide the sample fluid from the receiving tank to the reaction tanks; and an insert Components, used to be inserted into the plurality of reaction tanks of the inspection box, include: a plate-shaped component; and A plurality of tubes, arranged on the plate-shaped component, and arranged corresponding to the reaction tanks, wherein the tubes are configured to penetrate the inspection holes of the upper cover, and are inserted into the reaction tanks of the base, so that the sample fluids in the reaction tanks are retained in the tubes. The microfluidic cartridge as described in claim 1 further comprises: a first tilt angle, which is provided at the fluid connection between each reaction tank and each indicator tank, and tilts from each reaction tank to each indicator tank, so as to prevent the sample fluid from flowing back into each reaction tank; and a second tilt angle, which is provided at the fluid connection between each indicator tank and the collection tank, and tilts from each indicator tank to the collection tank, so as to prevent the sample fluid from flowing back into each indicator tank. The microfluidic cartridge as described in claim 1, wherein the reaction tanks and the indicator tanks are three in number and arranged in a row, and wherein: the fluid connection between the reaction tank located in the middle of the reaction tanks and the corresponding indicator tank is fan-shaped. The microfluidic cartridge as described in claim 1 further includes a sample cover that can be covered on the injection hole. The microfluidic cartridge as described in claim 1 further comprises a reaction tank cover for detachably covering the tubular objects of the insert. The microfluidic cartridge as described in claim 1 further includes at least one viewing window arranged corresponding to the indicator slots. The microfluidic cartridge described in claim 1 further includes a plurality of anti-overflow rings, which are respectively disposed on the upper edges of the inspection holes. A microfluidic cartridge as described in claim 1, wherein the bottom of the base corresponding to the plurality of reaction tanks protrudes more than the bottom around the base. A method for performing a test using a microfluidic cartridge as claimed in claim 1, comprising: (a) injecting a sample fluid into the base through the injection hole of the upper cover, wherein the sample fluid flows through the receiving tank through the drainage channels to the reaction tank, the indicator tank and the collection tank respectively; (b) inserting the plug-in through the upper cover into the base, wherein the tubes of the plug-in are respectively inserted into the reaction tanks of the base, so that the sample fluid is retained in the tubes; and (c) detecting the content of a test substance in the sample fluid in the tubes. The method as described in claim 9, wherein in step (c), a fluorescence value in the sample fluid in the tubes is detected by a fluorescence detection device.

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