Multi-bin paper-based micro-fluidic chip for visual detection of bacterial drug resistance

Abstract

The utility model discloses a multi-bin paper-based micro-fluidic chip for visual detection of bacterial drug resistance. The multi-bin paper-based micro-fluidic chip comprises a hydrophilic paper base, the detection mechanism is a hydrophilic channel established on the hydrophilic paper base; the detection mechanism comprises a sample adding bin and an anti-overflow bin, and the appearance of the anti-overflow bin is circular; one end of the anti-overflow channel is communicated with the sample adding bin, and the other end of the anti-overflow channel is communicated with the anti-overflow bin; the appearance of each reaction bin is an isosceles trapezoid; the plurality of reaction bins are uniformly distributed in a fan shape by taking the circle center of the sample adding bin as a central original point; one end of each diffusion channel is communicated with the reaction bin, and the other end of each diffusion channel is communicated with the sample adding bin; wherein the diffusion channels and the reaction bins are arranged in a one-to-one correspondence manner; the hydrophobic barrier structure is a hydrophobic area formed on the hydrophilic paper base through wax spraying printing along the outer edge of the detection mechanism. According to the multi-bin paper-based micro-fluidic chip for visual detection of bacterial drug resistance, provided by the utility model, rapid, high-throughput and low-cost detection of multiple bacterial drug resistance can be realized.

Description

Technical Field

[0001] This invention belongs to the field of microbial detection technology, and specifically relates to a multi-compartment paper-based microfluidic chip for visual detection of bacterial drug resistance. Background Technology

[0002] Traditional methods for detecting bacterial resistance, such as the broth dilution method, take 24-48 hours to complete. Automated antimicrobial susceptibility testing instruments cost over 500,000 yuan per unit and suffer from problems such as complex operation, high cost, and strong equipment dependence, making it difficult to meet the urgent needs of primary healthcare, emergency care, and animal husbandry for rapid screening of bacterial resistance. Therefore, against this backdrop, there is an urgent need to develop an innovative detection solution that is both rapid and economical, and highly adaptable, to overcome existing technological bottlenecks and improve the accessibility and accuracy of antimicrobial resistance testing. Utility Model Content

[0003] The purpose of this invention is to provide a multi-compartment paper-based microfluidic chip for visual detection of bacterial drug resistance, which can achieve rapid, high-throughput, and low-cost detection of bacterial multidrug resistance.

[0004] The technical solution provided by this utility model is as follows:

[0005] A multi-compartment paper-based microfluidic chip for visual detection of bacterial resistance includes:

[0006] Hydrophilic paper base;

[0007] The testing agency is responsible for establishing hydrophilic channels on the hydrophilic paper base;

[0008] The testing institutions include:

[0009] The sample loading chamber and spill prevention chamber are circular in shape.

[0010] An overflow prevention channel, one end of which is connected to the sample addition chamber, and the other end of which is connected to the overflow prevention chamber;

[0011] Multiple reaction chambers, each in the shape of an isosceles trapezoid, are evenly distributed in a fan shape with the center of the sample loading chamber as the origin.

[0012] Multiple diffusion channels, one end of which is connected to the reaction chamber and the other end of which is connected to the sample loading chamber;

[0013] The diffusion channels are configured in a one-to-one correspondence with the reaction chambers;

[0014] A hydrophobic barrier structure is a hydrophobic region formed by wax spraying on the hydrophilic paper substrate along the outer edge of the detection mechanism.

[0015] Preferably, the number of reaction chambers is seven; the number of diffusion channels is seven.

[0016] Preferably, one side of the trapezoidal upper bottom of the reaction chamber is connected to one end of the diffusion channel.

[0017] Preferably, the hydrophilic paper base has a size of 30 mm × 30 mm.

[0018] Preferably, the diameter of the sample loading chamber is 7 mm; the diameter of the anti-overflow chamber is 5 mm; the upper bottom length of the reaction chamber is 4 mm, the lower bottom length is 5 mm, and the height is 4 mm.

[0019] Preferably, the width of both the overflow prevention channel and the diffusion channel is 2 mm; the length of the diffusion channel is 6 mm, and the length of the overflow prevention channel is 4 mm.

[0020] Preferably, it further includes a transparent film covering the detection mechanism for sealing.

[0021] Preferably, the transparent film is made of polypropylene.

[0022] Preferably, the transparent film has a thickness of 50-100 μm, a light transmittance of ≥88%, a water vapor transmittance of 10-15 g / m² / day, and is coated with a quaternary ammonium salt antibacterial layer.

[0023] The beneficial effects of this invention are: the multi-compartment paper-based microfluidic chip for visual detection of bacterial drug resistance provided by this invention has a simple structure, requires fewer samples, and is easy to operate. It can improve the detection efficiency of bacterial multidrug resistance and achieve high-throughput, low-cost detection. Attached Figure Description

[0024] Figure 1 This is a schematic diagram of the multi-compartment paper-based microfluidic chip structure for visual detection of bacterial drug resistance as described in this utility model.

[0025] Figure 2 This invention relates the dilution factor of bacteria to fluorescence intensity when the multi-compartment paper-based microfluidic chip is used in experiments on bacterial multidrug resistance. Detailed Implementation

[0026] The present invention will now be described in further detail with reference to the accompanying drawings, so that those skilled in the art can implement it based on the description.

[0027] like Figure 1 As shown, this utility model provides a multi-compartment paper-based microfluidic chip for visual detection of bacterial drug resistance, comprising:

[0028] The hydrophilic paper base has hydrophilic properties; in this embodiment, the size of the hydrophilic paper base is 30 mm × 30 mm.

[0029] The testing mechanism comprises a hydrophilic channel established on the hydrophilic paper base; the interior of the testing mechanism is a hydrophilic region; the testing mechanism includes:

[0030] The sample dispensing chamber 110 and the overflow prevention chamber 120 are circular in shape; in this embodiment, the diameter of the sample dispensing chamber 110 is 7 mm; and the diameter of the overflow prevention chamber 120 is 5 mm.

[0031] An overflow prevention channel 121 is connected at one end to the sample loading chamber 110 and at the other end to the overflow prevention chamber 120; in this embodiment, the width of the overflow prevention channel 121 is 2 mm and the length is 4 mm.

[0032] The sample loading chamber 110 is used to add bacterial sample solution; the anti-overflow chamber 120 can be used as an anti-overflow device for bacterial sample solution, or it can be used to add bacterial sample solution, which is then diffused to the sample loading chamber 110 through the anti-overflow channel 121 by the capillary action of the hydrophilic paper base.

[0033] Multiple reaction chambers 130 are isosceles trapezoids in shape; the multiple reaction chambers 130 are evenly distributed in a fan shape with the center of the sample loading chamber 110 as the origin; different antibiotic diluents and rezakura dye are added to the multiple reaction chambers 130 respectively; in this embodiment, the number of reaction chambers 130 is seven; in this embodiment, the upper base of the reaction chamber 130 is 4 mm long, the lower base is 5 mm long, and the height is 4 mm;

[0034] Multiple diffusion channels 131 are provided, one end of which is connected to the trapezoidal upper bottom side of the reaction chamber 130, and the other end is connected to the sample loading chamber 110; in this embodiment, there are seven diffusion channels 131; the width of each diffusion channel 131 is 2 mm and the length is 6 mm.

[0035] The diffusion channel 131 is configured in a one-to-one correspondence with the reaction chamber 130.

[0036] The hydrophobic barrier structure 140 is a hydrophobic region formed by wax spraying on the hydrophilic paper substrate along the outer edge of the detection mechanism.

[0037] A transparent membrane is used to cover the testing mechanism for sealing; the transparent membrane is made of polypropylene (PP), with a thickness of 50-100μm, a light transmittance of ≥88%, a water vapor transmittance of 10-15g / m² / day, and a quaternary ammonium salt antibacterial layer coated on its surface.

[0038] like Figure 2As shown, the multi-compartment paper-based microfluidic chip for visual detection of bacterial drug resistance provided by this utility model is applied to the visual detection of bacterial multidrug resistance. The process of conducting a rapid screening experiment for bacterial multidrug resistance is as follows:

[0039] Step 1: Fabricate a porous chip using wax printing paper microfluidic technology: Print the hydrophobic barrier structure 140 onto the hydrophilic paper substrate according to the outer edge of the detection mechanism, forming a hydrophobic barrier on the hydrophilic paper substrate, thereby dividing the multi-compartment paper-based microfluidic chip for visual detection of bacterial resistance into a central sample area and an outer water storage area.

[0040] Step 2: Seven different common antibiotic diluents and resazurin dye are added to the seven reaction chambers 130 respectively. The antibiotics include ciprofloxacin, gentamicin, meropenem, cefazolin, ampicillin, tobramycin, and doxypenem. The concentration of the antibiotic diluents is optimized to more than 75 times the CLSI standard using a four-fold dilution method. The resazurin dye, as a redox indicator, has a colorimetric mechanism and fluorescence properties, and its fluorescence intensity is positively correlated with bacterial concentration. After gradient dilution of the bacteria, they are mixed with the resazurin dye at a 10:1 ratio. The metabolic activity of the bacteria simultaneously triggers changes in both the color and fluorescence intensity of the resazurin dye.

[0041] Step 3: Add E. coli solution to the sample loading chamber 110; the E. coli solution diffuses into the reaction chamber 130 through the capillary action of the hydrophilic paper base along the diffusion channel 131; incubate at 35°C, seal with the transparent membrane, and add water to maintain humidity, thereby promoting bacterial growth.

[0042] Step 4: Interpret the results based on the color change and fluorescence intensity of the reaction chamber 130: Based on the reduction reaction and fluorescence characteristics of the redox indicator resazurin, the color change of the resazurin dye is observed by the human eye, and the fluorescence change of the resazurin dye is recorded by a BV200 blue light transilluminator. The sensitivity of bacteria to antibiotics is determined by semi-quantitative analysis of the color change and fluorescence intensity within the reaction chamber 130: When bacteria are not inhibited by antibiotics, their metabolic activities will reduce the initially blue resazurin to resorufin, causing the color of the reaction chamber 130 to gradually change from blue to pink, indicating that the bacteria are resistant to the drug. The fluorescence intensity of the resazurin dye varies with the bacterial concentration; the higher the concentration, the stronger the fluorescence. If the antibiotic effectively inhibits bacterial growth, no bacterial metabolic reaction occurs, and the color of the reaction chamber 130 remains unchanged at blue with no significant fluorescence signal, indicating that the bacteria are sensitive. The less the color change, the more sensitive the bacteria.

[0043] Step 5: Combine the intelligent image analysis algorithm optimization of the mobile APP, and generate a quantitative drug resistance report that meets CLSI standards by analyzing the bacterial proliferation curve and color change threshold through time series analysis.

[0044] This invention provides a multi-compartment paper-based microfluidic chip for visual detection of bacterial drug resistance. It features a simple structure, low sample requirements, and easy operation, improving the detection efficiency of multidrug-resistant bacteria and achieving high-throughput, low-cost detection. The wax-printed microchannel design automates the detection process, enabling multi-antibiotic gradient testing with a single sample addition. Sample processing, reaction, and result reading are integrated into a single chip, reducing manual intervention. A single detection can cover multiple antibiotics, such as carbapenem-resistant Enterobacteriaceae classification, supporting multidrug resistance analysis and achieving high-throughput detection. The detection cycle is shortened to less than 10 hours, significantly improving detection speed and meeting the needs of rapid clinical decision-making. Only a small amount of bacterial sample is required, far less than traditional culture methods, enhancing detection convenience.

[0045] The multi-compartment paper-based microfluidic chip for visual detection of bacterial drug resistance provided by this utility model can be applied to rapid screening of bacterial multidrug resistance, which can reduce the detection cycle to 8-10 hours and the cost of a single test to 90 yuan / test, solving technical pain points such as short window period for accurate medication in emergency departments and lack of constant temperature equipment in primary medical institutions.

[0046] Although the embodiments of this utility model have been disclosed above, they are not limited to the applications listed in the specification and embodiments. They can be applied to various fields suitable for this utility model. For those skilled in the art, other modifications can be easily made. Therefore, without departing from the general concept defined by the claims and their equivalents, this utility model is not limited to the specific details and the illustrations shown and described herein.

Claims

1. A multi-compartment paper-based microfluidic chip for visual detection of bacterial drug resistance, characterized in that, include: Hydrophilic paper base; The testing agency is responsible for establishing hydrophilic channels on the hydrophilic paper base; The testing institutions include: The sample loading chamber and spill prevention chamber are circular in shape. An overflow prevention channel, one end of which is connected to the sample addition chamber, and the other end of which is connected to the overflow prevention chamber; Multiple reaction chambers, each in the shape of an isosceles trapezoid, are evenly distributed in a fan shape with the center of the sample loading chamber as the origin. Multiple diffusion channels, one end of which is connected to the reaction chamber and the other end of which is connected to the sample loading chamber; The diffusion channels are configured in a one-to-one correspondence with the reaction chambers; A hydrophobic barrier structure is a hydrophobic region formed by wax spraying on the hydrophilic paper substrate along the outer edge of the detection mechanism.

2. The multi-compartment paper-based microfluidic chip for visual detection of bacterial resistance according to claim 1, characterized in that, The number of reaction chambers is seven; the number of diffusion channels is seven.

3. The multi-compartment paper-based microfluidic chip for visual detection of bacterial resistance according to claim 1, characterized in that, The upper bottom side of the trapezoidal structure of the reaction chamber is connected to one end of the diffusion channel.

4. The multi-compartment paper-based microfluidic chip for visual detection of bacterial resistance according to claim 1, characterized in that, The hydrophilic paper base has dimensions of 30 mm × 30 mm.

5. The multi-compartment paper-based microfluidic chip for visual detection of bacterial resistance according to claim 1, characterized in that, The diameter of the sample loading chamber is 7 mm; the diameter of the spill prevention chamber is 5 mm; the upper bottom of the reaction chamber is 4 mm long, the lower bottom is 5 mm long, and the height is 4 mm.

6. The multi-compartment paper-based microfluidic chip for visual detection of bacterial resistance according to claim 1, characterized in that, The width of both the overflow prevention channel and the diffusion channel is 2 mm; the length of the diffusion channel is 6 mm, and the length of the overflow prevention channel is 4 mm.

7. The multi-compartment paper-based microfluidic chip for visual detection of bacterial resistance according to claim 1, characterized in that, Also includes: A transparent film is used to cover the detection mechanism for sealing.

8. The multi-compartment paper-based microfluidic chip for visual detection of bacterial resistance according to claim 7, characterized in that, The transparent film is made of polypropylene.

9. The multi-compartment paper-based microfluidic chip for visual detection of bacterial resistance according to claim 8, characterized in that, The transparent film has a thickness of 50-100 μm, a light transmittance of ≥88%, and a water vapor transmission rate of 10-15 g / m². 2 / day, with a quaternary ammonium salt antibacterial layer coated on the surface.

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