CN120629086A - Wireless test strip analyzer and method of use thereof - Google Patents

Abstract

The present invention discloses a wireless test strip analyzer and its use method, relating to the field of biological detection technology. The analyzer comprises a circuit board, a test strip slot, a battery, and a protective case. The circuit board, test strip slot, and battery are built into the protective case, and the battery is connected to the circuit board. The test strip slot is used to hold a test strip. The test strip is dripped with an amplification product based on a biomimetic dandelion isothermal amplification system for detecting the number of CTCs. The present invention can improve the sensitivity of CTC detection, reduce detection costs, and reduce reliance on expensive equipment.

Description

Wireless test strip analyzer and use method thereof

Technical Field

The invention relates to the technical field of biological detection, in particular to a wireless test strip analyzer and a using method thereof.

Background

Detection of Circulating Tumor Cells (CTCs) is critical, but faces challenges such as low sensitivity, high cost, and complex operation. The existing CTCs detection method lacks sufficient sensitivity, and the high-precision CTCs detection relies on expensive advanced instruments, such as a single-cell sequencing platform and microfluidic chip equipment, so that the cost is high, the instruments are complex, and the wide application is not facilitated.

Disclosure of Invention

The invention aims to provide a wireless test strip analyzer and a using method thereof, which aim to solve or improve at least one of the technical problems.

In order to achieve the above object, the present invention provides the following solutions:

The wireless test strip analyzer comprises a circuit board, a test strip clamping groove, a battery and a protective shell, wherein the circuit board, the test strip clamping groove and the battery are arranged in the protective shell and are connected with the circuit board, the test strip clamping groove is used for placing a test strip, and an amplification product based on a bionic dandelion isothermal amplification system is dripped on the test strip and used for detecting the number of CTCs.

Optionally, the circuit board integrates a 365nm ultraviolet LED, an XYZ true color sensor, a bluetooth 5.1 system on chip, and a 3.3 volt power supply.

Optionally, the bluetooth 5.1 system-on-chip is further wirelessly connected with a smart phone of the user, and is configured to receive a wireless instruction sent by the smart phone of the user and transmit detection information.

Optionally, the bionic dandelion-based isothermal amplification system consists of hexapod DNAWALKER, a nonlinear DNA self-assembly technology and an asymmetric carrier with high signal probe loading efficiency, wherein the asymmetric carrier with high signal probe loading efficiency adopts AuFe Janus nano particles.

The invention also provides a using method of the wireless test strip analyzer, which comprises the following steps of:

After the analyzer is started, bluetooth connection is established between the analyzer and the intelligent mobile phone through the BLE MCU, and the intelligent mobile phone sends a control instruction to the BLE MCU in a wireless mode;

After receiving the instruction, the BLE MCU sets the specific GPIO pin high, activates the NMOS transistor to be conducted, and thus the ultraviolet LED is lightened;

The XYZ true color sensor is communicated with the XYZ true color sensor through an I2C protocol to complete sensor parameter configuration and data acquisition, converts a captured optical signal into an XYZ tristimulus value based on an international illumination committee 1931 color space standard to realize standardized measurement of fluorescence intensity, and is also internally provided with a signal processing unit to directly transmit the processed digital optical signal to the BLE MCU through an I2C interface;

After the measurement of color data is completed, the BLE MCU controls the sensor to enter a power-down mode, and the ultraviolet LED is turned off to reduce the power consumption;

And (3) returning the result, namely returning the collected XYZ data to the smart phone through Bluetooth by the BLE MCU, and evaluating the fluorescence intensity of the T-line of the test strip by the user based on the received XYZ values.

According to the specific embodiment provided by the invention, the invention discloses the following technical effects:

The invention discloses a wireless test strip analyzer and a use method thereof, wherein the analyzer comprises a circuit board, a test strip clamping groove, a battery and a protective shell, wherein the circuit board, the test strip clamping groove and the battery are arranged in the protective shell and are connected with the circuit board, the test strip clamping groove is used for placing a test strip, and an amplification product based on a bionic dandelion isothermal amplification system is dripped on the test strip and used for detecting the quantity of CTCs. The invention can improve the sensitivity of CTCs detection, reduce the detection cost and reduce the dependence on expensive instruments.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions of the prior art, the drawings that are needed in the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

FIG. 1 is a schematic diagram of a wireless test strip analyzer according to the present invention.

FIG. 2 is a schematic circuit diagram of a wireless test strip analyzer according to the present invention.

FIG. 3 shows the effect of MCF-7 cells with different concentrations in the present embodiment, wherein (a) part is a real image of the detection of MCF-7 cells with different concentrations (0, 10, 100, 1000 cells/mL) and (b) part is a T-line fluorescence quantitative bar chart of the wireless test strip analyzer of the present invention for the test strips of MCF-7 cells with different concentrations.

Fig. 4 is a detailed schematic diagram of the wireless test strip analyzer in this embodiment.

Detailed Description

The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

The invention aims to provide a wireless test strip analyzer and a using method thereof, which aim to solve or improve at least one of the technical problems.

In order that the above-recited objects, features and advantages of the present invention will become more readily apparent, a more particular description of the invention will be rendered by reference to the appended drawings and appended detailed description.

The invention provides a wireless test strip analyzer, which comprises a circuit board, a test strip clamping groove, a battery and a protective shell, wherein the circuit board, the test strip clamping groove and the battery are arranged in the protective shell and are connected with the circuit board, the test strip clamping groove is used for placing a test strip, and an amplification product based on a bionic dandelion isothermal amplification system is dripped on the test strip and used for detecting the number of CTCs.

The bionic dandelion isothermal amplification system consists of hexapod DNAWALKER, a nonlinear DNA self-assembly technology and an asymmetric carrier with high signal probe loading efficiency, namely AuFe Janus nano particles, and has remarkable amplification efficiency. Compared with the traditional isothermal amplification system, the amplification efficiency of the bionic dandelion isothermal amplification system is improved by about 6.72 times.

As a specific implementation mode, the detection system mainly comprises a bionic dandelion isothermal amplification system, a lateral flow immunochromatography test strip and a wireless test strip analyzer. First, CTCs are enriched and isolated from red cell lysed blood using a magnetic bead labeled anti-EpCAM monoclonal antibody under the influence of a magnetic field. Then, adding a bionic dandelion isothermal amplification system. CTCs can competitively bind to the aptamer, releasing the aptamer-blocked hexapod DNAWALKER. With the help of Mg ²⁺, hexapod DNAWALKER is used as DNAzyme, specifically cuts RNA base of D-RNA, and efficiently releases dandelion seed-shaped DNA macromolecules. This process is very similar to the natural phenomenon of dandelion floss scattering, so the system is named as a bionic dandelion isothermal amplification system. As an amplification product of the bionic dandelion isothermal amplification system, the bionic dandelion seeds are dripped on a sample pad of the test strip after magnetic separation. After 15 minutes, the fluorescence intensity was measured, and accurate detection of the number of CTCs was achieved by establishing a positive correlation between the number of CTCs and the fluorescence intensity.

As a more specific embodiment, a wireless test strip analyzer as shown in fig. 1 is constructed, which is mainly composed of four parts, a highly integrated circuit board, a test strip card slot, a battery, and a protective case (fig. 1). The circuit board integrates a 365nm ultraviolet LED, XYZ true color sensor, bluetooth 5.1 system on chip and 3.3 volt power supply (fig. 1). The user controls the fluorescence detector through a wireless instruction sent by the smart phone. After receiving the instruction, the Bluetooth chip system can activate the ultraviolet LED and the fluorescence sensor to measure the fluorescence intensity of the T-line of the test strip. The sensor then captures the fluorescence information and outputs it as X, Y and Z values.

The wireless LFIA test paper analyzer uses an XYZ true color sensor to determine the concentration of CTCs by analyzing the fluorescence intensity of the T line on the LFIA test paper under uv light. The specific working mechanism is as follows:

1. Bluetooth connection and instruction transmission

After the power-on, the analyzer establishes Bluetooth connection with the smart phone through a Bluetooth low energy micro controller unit (BLE MCU, model HJ-531 IMH). The smart phone wirelessly transmits the control instruction to the MCU.

2. Ultraviolet light source activation

When the BLE MCU receives the instruction, a General Purpose Input Output (GPIO) pin is set to a high level, and an N-channel Metal Oxide Semiconductor Field Effect Transistor (MOSFET) is triggered, so that the ultraviolet LED is turned on.

3. Sensor communication and data acquisition

Then, the BLE MCU establishes communication with the XYZ true color sensor (model AS 73211) through the I2C protocol, and sensor parameter configuration and data acquisition are completed.

The sensor has the core function of converting the captured light signals into hexadecimal values according to the 1931 color space standard of the international committee of illumination, so as to realize the accurate quantification of the fluorescence intensity. The integrated signal processing unit directly transmits the processed digital optical signals to the BLE MCU through the I2C interface.

4. Low power mode initiation

After the data acquisition is completed, the BLE MCU instructs the color sensor to enter a power-down state, and the ultraviolet LED is turned off, so that the power consumption is reduced.

5. Data wireless transmission and result analysis

Finally, the collected hexadecimal values are transmitted to the smart phone through Bluetooth in a wireless mode, and a user can correlate the fluorescence intensity with the CTCs concentration through the data.

By means of the design, the analyzer realizes accurate and efficient fluorescence detection by means of advanced components such as an XYZ true color sensor and an optimized signal processing technology, and accuracy of result interpretation is ensured.

Subsequently, the feasibility of wireless cell sensors based on wireless LFIA test strip analyzers was assessed. The feasibility of wireless cell sensors was verified by detecting MCF-7 cells at concentrations of 0,10,100 and 1000 cells (FIG. 3). As a result of the verification, the fluorescence intensity of the test strip T-line was proportional to the increase in the cell concentration (FIG. 3 (a)). Whereas no distinct fluorescence bands were observed on the T-line of the blank samples. The dipstick is then inserted into a wireless dipstick analyzer for more accurate quantification (fig. 3 (b)). The detection result obtained from the wireless test strip analyzer is consistent with the visual interpretation result of the test strip, and also provides accurate numerical values. These findings indicate that the wireless dipstick analyzer has good feasibility, and can ensure accurate detection of CTCs.

Conclusion(s)

In the embodiment, a novel wireless cell sensor combining a bionic dandelion isothermal system, a test strip platform and a wireless test strip analyzer is successfully developed. Based on the bionics principle, a bionic dandelion isothermal system is designed to improve the amplification efficiency. Due to the synergistic effect of the high CuInS2@ZnS quantum dot load rate of the bionic dandelion seeds, the fluorescent signal probe load rate of AuFe JNPs and the high digestion efficiency of hexapod DNAWALKER, the amplification efficiency of the bionic dandelion isothermal system is 6.72 times higher than that of the traditional isothermal amplification system. The application of the test strip platform makes the operation simpler and more convenient. The cell sensor exhibits excellent performance in terms of sensitivity, specificity and reproducibility. In particular, the ease of operation, makes such wireless cell sensors advantageous in primary healthcare and large-scale screening. It is worth noting that clinical sample experiments show that the method has higher accuracy and anti-interference capability, and can be used for detecting CTCs in whole blood samples. In view of this, newly developed wireless cell sensors have great potential in the clinical detection of CTCs.

As another specific embodiment, a method of using the wireless test strip-based analyzer will be described in detail.

The wireless test strip analyzer adopts an XYZ true color sensor, and the concentration of CTCs is determined by analyzing the fluorescence intensity of T-line on the test strip under ultraviolet light. The specific operation steps are as follows:

1. And (3) starting and initializing, namely after the analyzer is started, establishing Bluetooth connection with the smart phone through a BLE MCU (model HJ-531 IMH). The mobile phone sends a control instruction to the BLE MCU in a wireless mode.

2. And activating the ultraviolet LED, namely after the BLE MCU receives the instruction, setting a specific GPIO pin high, and activating the NMOS transistor to be conducted, so that the ultraviolet LED is lightened.

And 3, communication between the XYZ true color sensor and the BLE MCU through an I2C protocol, and establishing communication between the BLE MCU and the XYZ true color sensor (model AS 73211) to complete sensor parameter configuration and data acquisition. The sensor converts the captured light signals to XYZ values based on the international commission on illumination (CIE) 1931 color space standard. The set of values (X, Y, Z) can accurately represent the perception of the human eyes to the color, and standardized measurement of the fluorescence intensity is realized. The sensor is internally provided with a signal processing unit, and the processed digital optical signals can be directly transmitted to the BLE MCU through the I2C interface.

4. And after the measurement of the color data is completed, the BLE MCU controls the sensor to enter a power-down mode, and the ultraviolet LEDs are turned off to reduce the power consumption.

5. And the BLE MCU finally transmits the collected XYZ data back to the smart phone through Bluetooth. The user evaluates the fluorescence intensity of the T-line of the test strip based on the received XYZ values.

The design adopts advanced components such as an XYZ true color sensor and the like, and combines an optimized signal processing algorithm, thereby ensuring the accuracy and the high efficiency of fluorescence detection and providing a reliable basis for judging the test strip.

The wireless test strip analyzer uses 365nm ultraviolet LEDs to excite fluorescent substances on the test paper, and analyzes effective information expressed by the test paper by detecting fluorescence intensity. The user controls the operation of the fluorescence detector through a wireless instruction sent by the smart phone. After receiving the instruction, the Bluetooth 5.1 system-on-chip activates the ultraviolet LED and the XYZ true color sensor to measure the fluorescence on the test paper. Specifically, after the ultraviolet light excites the fluorescent substance, the sensor captures color information of the fluorescence reflection and outputs it in the form of X, Y, Z values in the XYZ color space. The sensor transmits data to the Bluetooth 5.1 system level chip through the I2C interface, and after the data are processed, the detection result is wirelessly transmitted to the intelligent mobile phone through Bluetooth. The ultraviolet LED is directly powered by a battery to provide sufficient driving current, so that the light power of ultraviolet light can be ensured to effectively excite fluorescent substances. Meanwhile, the XYZ true color sensor and the Bluetooth 5.1 system-on-chip are powered by a 3.3V power supply regulated by a low dropout regulator (LDO) to ensure the normal operation of the XYZ true color sensor and the Bluetooth 5.1 system-on-chip. The circuit diagram of the wireless test strip analyzer is shown in fig. 2, and the corresponding detailed circuit schematic diagram is shown in fig. 4. The wireless test strip analyzer realizes a complete functional chain of fluorescence excitation, color detection and wireless data transmission, and is suitable for portable fluorescence detection application scenes.

In the present specification, each embodiment is described in a progressive manner, and each embodiment is mainly described in a different point from other embodiments, and identical and similar parts between the embodiments are all enough to refer to each other.

The principles and embodiments of the present invention have been described herein with reference to specific examples, which are intended to facilitate an understanding of the principles and embodiments of the invention and are to be varied in scope and detail by persons skilled in the art in light of the teachings of the invention. In view of the foregoing, this description should not be construed as limiting the invention.

Claims (5)

1. A wireless test strip analyzer is characterized by comprising a circuit board, a test strip clamping groove, a battery and a protective shell, wherein the circuit board, the test strip clamping groove and the battery are arranged in the protective shell and are connected with the circuit board, the test strip clamping groove is used for placing a test strip, and an amplification product based on a bionic dandelion isothermal amplification system is dripped on the test strip and used for detecting the number of CTCs.

2. The wireless test strip analyzer of claim 1, wherein the circuit board integrates a 365nm ultraviolet LED, an XYZ true color sensor, a bluetooth 5.1 system on chip, and a 3.3 volt power supply.

3. The wireless test strip analyzer of claim 2, wherein the bluetooth 5.1 system-on-chip is further wirelessly connected to a smart phone of the user for receiving wireless instructions sent by the smart phone of the user and transmitting detection information.

4. The wireless test strip analyzer of claim 1, wherein the bionic dandelion-based isothermal amplification system comprises hexapod DNAWALKER, a nonlinear DNA self-assembly technology and an asymmetric carrier with high signal probe loading efficiency, and the asymmetric carrier with high signal probe loading efficiency adopts AuFe Janus nano particles.

5. A method of using a wireless test strip analyzer, employing the analyzer of any one of claims 1-4, comprising:

After the analyzer is started, bluetooth connection is established between the analyzer and the intelligent mobile phone through the BLE MCU, and the intelligent mobile phone sends a control instruction to the BLE MCU in a wireless mode;

After receiving the instruction, the BLE MCU sets the specific GPIO pin high, activates the NMOS transistor to be conducted, and thus the ultraviolet LED is lightened;

The XYZ true color sensor is communicated with the XYZ true color sensor through an I2C protocol to complete sensor parameter configuration and data acquisition, converts a captured optical signal into an XYZ tristimulus value based on an international illumination committee 1931 color space standard to realize standardized measurement of fluorescence intensity, and is also internally provided with a signal processing unit to directly transmit the processed digital optical signal to the BLE MCU through an I2C interface;

After the measurement of color data is completed, the BLE MCU controls the sensor to enter a power-down mode, and the ultraviolet LED is turned off to reduce the power consumption;

And (3) returning the result, namely returning the collected XYZ data to the smart phone through Bluetooth by the BLE MCU, and evaluating the fluorescence intensity of the T-line of the test strip by the user based on the received XYZ values.