CN-121975608-A - Integrated centrifugal PCR fluid verification and calibration module

CN121975608ACN 121975608 ACN121975608 ACN 121975608ACN-121975608-A

Abstract

The invention discloses an integrated centrifugal PCR fluid verification and calibration module, and relates to the technical field of calibration of centrifugal microfluidic PCR devices. The module comprises a module main body, wherein the module main body is arranged on an output shaft of a centrifugal driving mechanism of the centrifugal microfluidic PCR device, the module main body is formed by sequentially connecting a top layer PCB, a fluid verification chip, a calibration chip and a bottom layer PCB in a positioning and overlapping manner, and a four-probe temperature detection unit, a pressure detection unit, a fluorescence simulation unit, a fluid verification unit, a control unit and a battery are integrated on the module main body. The invention can stably supply power under the centrifugal working condition, synchronously finish the calibration of temperature, pressure and fluorescence parameters and the functional verification of flow channel fluid, and provide a reliable technical scheme for the accurate and efficient calibration of the PCR device.

Inventors

ZENG YOUHONG
LIU LINGXUAN
MA XINGWEI
QIN XUYING
ZHANG HANXING
LI QI
Yuan Yatan
WANG LEI
CHEN HAO
ZHANG YIZE
DING YINGYING

Assignees

北京亿美诺生物科技股份有限公司

Dates

Publication Date: 20260505
Application Date: 20260202

Claims (8)

1. The integrated centrifugal PCR fluid verification and calibration module is characterized by comprising a module main body, wherein the module main body is arranged on an output shaft of a centrifugal driving mechanism of a centrifugal microfluidic PCR device, the module main body is disc-shaped and is formed by positioning and superposing a top layer PCB (1), a fluid verification chip (3), a calibration chip (4) and a bottom layer PCB (6) in sequence, the module main body is integrated with a four-probe temperature detection unit, a pressure detection unit, a fluorescence simulation unit, a fluid verification unit, a control unit and a battery (7), the four-probe temperature detection unit, the pressure detection unit, the fluorescence simulation unit and the fluid verification unit are electrically connected with the control unit, the battery provides power for the whole module main body, the four-probe temperature detection unit collects temperature data of the PCR device and sends the temperature data of the PCR device to the control unit, the pressure detection unit collects pressure data of the PCR device and sends the pressure data to the control unit, the fluorescence simulation unit provides standard fluorescence for a fluorescent module of the PCR device, the fluid verification unit simulates a flow channel environment of the PCR device under the centrifugal force, the control unit sends received data to an upper computer, the upper computer processes the received data and sends instructions, and the control unit receives the instructions to control the module main body to complete temperature, the pressure, the fluorescence flow channel, and the fluid verification and the working parameters of the PCR device.
2. The integrated centrifugal PCR fluid verification and calibration module according to claim 1, wherein the fluid verification unit comprises a microfluidic flow channel structure, a chamber, a valve, a white light lamp (37) and a small photoelectric sensor (39), the microfluidic flow channel structure, the chamber and the valve are arranged on the fluid verification chip (3), light rays emitted by the white light lamp (37) downwards irradiate on the fluid verification chip (3) and pass through the flow channel structure of the fluid verification chip (3), finally irradiate on the small photoelectric sensor (39) below, the small photoelectric sensor (39) completes collection of optical signals and transmits the optical signals to the control unit, and flow and distribution states of fluid in the fluid verification chip (3) are monitored in real time, so that accurate verification of the flow channel fluid functions of the centrifugal microfluidic PCR device is realized.
3. An integrated centrifugal PCR fluid verification and calibration module according to claim 1, wherein the four-probe temperature detection unit comprises four temperature sensors, four heating chambers (32) are arranged in an array on the calibration chip (4), and the four temperature sensors are arranged in the four heating chambers (32).
4. An integrated centrifugal PCR fluid verification and calibration module according to claim 1, characterized in that the pressure detection unit comprises a pressure sensor (46), a pressure sensor fixing groove (25) is arranged on the calibration chip (4), the pressure sensor (46) is arranged in the pressure sensor fixing groove (25), and the pressure sensor (46) collects pressure data in the extrusion process in real time when the calibration chip (4) is compressed.
5. The integrated centrifugal PCR fluid verification and calibration module according to claim 1, wherein the fluorescence simulation unit comprises four fluorescent lamps (33), a light equalizing plate (8), a fluorescence calibration channel (15), a condensing lens (34), an optical filter (35) and a large photoelectric sensor (36), the four fluorescent lamps (33) are arranged in the fluorescence calibration channel (15), light rays emitted by the fluorescent lamps (33) are focused through the condensing lens (34) after being homogenized by the light equalizing plate (8), specific wavelengths are screened by the optical filter (35), and are secondarily focused through another group of condensing lenses (34), and finally the light rays are transmitted to the large photoelectric sensor (36) to finish signal receiving.
6. The integrated centrifugal PCR fluid verification and calibration module according to claim 1, wherein the control unit comprises a main control chip, an AD conversion chip, a Bluetooth module and a battery power supply control module, the AD conversion chip receives and converts data signals output by the calibration chip (4), the fluid verification chip (3), the upper layer PCB (1) and the lower layer PCB (6), the converted data signals are transmitted to the Bluetooth module, the Bluetooth module transmits the received data signals to the main control chip for processing, the processed signal data are transmitted to an upper computer, and the upper computer processes the data and sends out control commands to control the whole calibration module to perform calibration and verification.
7. The integrated centrifugal PCR fluid verification and calibration module in accordance with claim 6, wherein the master control chip is an STM32F103 microcontroller, an STM32L476 controller, or an ESP32 controller, and the AD conversion chip is an ADs1296 AD converter, an ADs1118 converter, or an LTC2499 converter.
8. An integrated centrifugal PCR fluid verification and calibration module according to claim 1, characterized in that an upper layer spacer (2) is provided between the top layer PCB (1) and the fluid verification chip (3) and a lower layer spacer (5) is provided between the calibration chip (4) and the bottom layer PCB (6).

Description

Integrated centrifugal PCR fluid verification and calibration module Technical Field The invention belongs to the technical field of calibration of centrifugal microfluidic PCR devices, and particularly relates to an integrated centrifugal PCR fluid verification and calibration module. Background Microfluidic technology has been widely used in various fields such as biomedical detection, clinical diagnosis, environmental monitoring, etc. by virtue of the core advantages of miniaturization, integration, and low consumption. The centrifugal microfluidic PCR (Polymerase Chain Reaction ) device does not need complex pump valve driving, and fluid transportation and reaction regulation and control in the micro-channel can be realized only by virtue of centrifugal force, so that the centrifugal microfluidic PCR (Polymerase Chain Reaction ) device becomes the preferred equipment for on-site rapid detection scenes. The core accuracy of PCR detection depends on the temperature control accuracy (denaturation, annealing, extension three-stage temperature accuracy), pressure stability (the adaptation accuracy of chips with different thickness and double Peltier clamp heating), and fluorescence detection sensitivity (target nucleic acid quantitative accuracy). Therefore, in the links of factory calibration, daily maintenance and periodic calibration of the device, the parameters need to be accurately calibrated. Meanwhile, the micro-flow channel is used as a core carrier for fluid transportation and reaction, the on-off state and the fluid flow stability of the micro-flow channel directly influence the reliability of a detection result, and the functional verification of the fluid in the flow channel also becomes a key link of a calibration flow. Along with the continuous improvement of the requirements of clinical diagnosis on detection precision and efficiency, the calibration requirement of a centrifugal microfluidic PCR device is urgent, and research and development of an integrated technical scheme which is adaptive to the centrifugal working characteristics and can realize multi-parameter synchronous calibration and fluid verification is urgent. However, the development of the calibration technology of the microfluidic PCR device is delayed from the iteration speed of the device itself, and the existing calibration scheme is difficult to adapt to the special requirement of the centrifugal working condition, which becomes a core bottleneck for restricting the accurate application of the device. Disclosure of Invention The invention aims to solve the problems that the existing calibration tool is independent in structure, cannot adapt to a centrifugal working environment, is difficult to synchronously complete temperature, pressure and fluorescence parameter calibration and flow channel fluid function verification in a centrifugal state, and causes redundant flow and poor adaptability, and the problems that the existing calibration tool is easy to wind and fall off due to wired power supply under a centrifugal rotation working condition, and further causes interruption of the calibration process or out-of-control precision are solved. Meanwhile, through the creative structural design, the integrated operation of stable power supply, multi-parameter synchronous accurate calibration and runner fluid function real-time verification under the centrifugal working condition is realized, the calibration efficiency and the suitability of the centrifugal microfluidic PCR device are improved, and a reliable technical scheme is provided for the accurate and efficient calibration of the device. The technical scheme includes that the integrated centrifugal PCR fluid verification and calibration module comprises a module main body, wherein the module main body is arranged on an output shaft of a centrifugal driving mechanism of a centrifugal microfluidic PCR device, the module main body is formed by positioning and superposing and connecting a top layer PCB, a fluid verification chip, a calibration chip and a bottom layer PCB in sequence, four probe temperature detection units, a pressure detection unit, a fluorescence simulation unit, a fluid verification unit, a control unit and a battery are integrated on the module main body, the four probe temperature detection units, the fluorescence simulation unit and the fluid verification unit are electrically connected with the control unit, the battery provides power for the whole module main body, the four probe temperature detection units acquire temperature data of the PCR device and send the temperature data of the PCR device to the control unit, the pressure detection unit acquires pressure data of the PCR device and sends the pressure data to the control unit, the fluorescence simulation unit provides standard fluorescence for a fluorescent module of the PCR device, the fluid verification unit simulates a flow channel environment of the PCR device under the centrifugal force, the