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US-12618103-B2 - Instrument and method for extracting and detecting nucleic acids

US12618103B2US 12618103 B2US12618103 B2US 12618103B2US-12618103-B2

Abstract

Provided is a device for extracting and detecting nucleic acids, comprising one or more sample receiving modules, one or more lysing modules, one or more extraction module of nucleic acids, one or more amplification module of nucleic acids, and a detection module. Also provided is a molecular in vitro diagnostic instrument comprising the device for extracting and detecting nucleic acids and an automatic control system. Further provided is an in vitro diagnosis method by means of performing extraction and amplification of nucleic acids in a sample using the device for extracting and detecting nucleic acids.

Inventors

  • Xing Yang
  • Xianhua Wang
  • Yang Song
  • Jianxin Ye
  • Wei Zhang

Assignees

  • JIAXING ACCUNOME BIOTECHNOLOGY CO., LTD.

Dates

Publication Date
20260505
Application Date
20200908
Priority Date
20190910

Claims (20)

  1. 1 . A device for extracting and detecting nucleic acids, comprising: one or more sample receiving modules, each sample receiving module comprises a chamber adapted to receive a sample or sample container; one or more lysing modules, each lysing module comprises a chamber adapted to receive a lysing reagent or a lysing kit; one or more nucleic acid extraction modules, each nucleic acid extraction module of nucleic acids comprises a chamber adapted to receive nucleic acid extraction reagents or a nucleic acid extraction kit; one or more nucleic acid amplification modules, each nucleic acid amplification module comprises a plurality of chambers adapted to receive nucleic acid amplification reagents or an amplification tube of a nucleic acid amplification kit, wherein the one or more nucleic acid amplification modules further comprise a capping mechanism to close a cap of the amplification chamber or amplification tube, the capping mechanism comprising a cap assembly and a motion assembly, wherein the cap assembly comprises a cap clamping plate and a stepping motor, wherein the stepping motor controls up and down movements of the cap clamping plate, wherein the motion assembly is configured to move back and forth in the horizontal direction, wherein the amplification chamber of the nucleic acid amplification module is connected to the motion assembly, wherein the capping mechanism is configured so that: when the amplification chamber or the amplification tube moves with the motion assembly to be close to the cap clamping plate, the stepping motor drives the cap clamping plate to move up, lifts the cap of the amplification chamber or the amplification tube, and flips the cap; and when the cap is lifted to an appropriate angle, the cap clamping plate no longer moves upward, the motion assembly continues to move toward the cap clamping plate, and then the stepping motor drives the clamping plate to move downward to press the cap; wherein the device further comprises a detection module.
  2. 2 . The device of claim 1 , wherein the lysing module is configured to lyse the sample by means of a lysing reagent, an enzyme, ultrasound or physical grinding.
  3. 3 . The device of claim 1 , wherein the lysing module further comprises an oscillator composed of an eccentric shaft, an eccentric seat, and a motor, wherein the eccentric shaft rotates around an axis of the motor, causing the eccentric seat to oscillate.
  4. 4 . The device of claim 3 , wherein the lysing module is adapted to receive grinding particles or a lysing kit comprising grinding particles to physically grind the sample.
  5. 5 . The device of claim 4 , wherein the lysing kit comprises a lysing tube for receiving the grinding particles.
  6. 6 . The device of claim 1 , wherein the nucleic acid extraction module comprises one or more nucleic acid extraction units, wherein each of the one or more nucleic acid extraction units is composed of a binding unit, a washing unit, and an eluting unit.
  7. 7 . The device of claim 6 , further comprising a dispensing tube that is configured to go into a binding unit, a washing unit, and an eluting unit of the nucleic acid extraction module, wherein the nucleic acid extraction module is configured to: combine the sample with magnetic beads in a solution in a container, whereby nucleic acids in the sample are bound by the magnetic beads; insert the tip of the dispensing tube into the bottom of the container; draw the solution containing the magnetic beads into the dispensing tube, whereby a magnet close to the dispensing tube adsorbs the magnetic beads to a side wall of the dispensing tube; dispense the solution from the dispensing tube; move the dispensing tube to another container; and move the magnet relative to the dispensing tube such that the effect of the magnetic field disappears and the magnetic beads are released from the side wall of the dispensing tube.
  8. 8 . The device of claim 6 , wherein the nucleic acid amplification module of comprises a plurality of nucleic acid amplification units, wherein each of the nucleic acid amplification units corresponds to a nucleic acid extraction sample obtained by a nucleic acid extraction unit in the nucleic acid extraction module, wherein each nucleic acid amplification unit comprises two or more chambers that are configured to independently perform amplification reactions.
  9. 9 . The device of claim 1 , wherein the nucleic acid extraction module comprises a magnet that is configured to apply a magnetic field to control a magnetic material and/or a nucleic acid-binding magnetic material.
  10. 10 . The device of claim 9 , further comprising a dispensing tube that is configured to go into a binding unit, a washing unit, and an eluting unit of the nucleic acid extraction module, wherein the nucleic acid extraction module is configured to: combine the sample with magnetic beads in a solution in a container, whereby nucleic acids in the sample are bound by the magnetic beads; insert the tip of the dispensing tube into the bottom of the container; draw the solution containing the magnetic beads into the dispensing tube, whereby the magnet close to the dispensing tube absorbs the magnetic beads to a side wall of the dispensing tube, dispense the solution from the dispensing tube; move the dispensing tube to another container; move the magnet relative to the dispensing tube such that the effect of the magnetic field disappears and the magnetic beads are released from the side wall of the dispensing tube.
  11. 11 . The device of claim 1 , wherein the nucleic acid amplification module is configured for isothermal amplification or polymerase chain reaction (PCR).
  12. 12 . The device of claim 1 , wherein the nucleic acid amplification module further comprises a temperature regulator configured to independently heat or cool each of the plurality of chambers.
  13. 13 . The device of claim 1 , wherein the detection module is configured to detect labels carried by nucleic acids, and wherein the labels are identifiable by fluorescence, chemiluminescence, bioluminescence, radioluminescence, electroluminescence, electrochemiluminescence, mechanoluminescence, crystalline luminescence, thermoluminescence, sonoluminescence, phosphorescence, photoluminescence, enzymatic reactions, or radioactivity.
  14. 14 . The device of claim 1 , further comprising a liquid dispensing module to transfer or dispense samples or reagents in the device between two or more locations, the liquid dispensing module comprises: one or more sensors; one or more dispensing heads; and a Cartesian coordinate robot that provides 3-axis linear movement for the dispensing heads.
  15. 15 . The device of claim 1 , further comprising a reagent and consumable loading area comprising one or more of a pipette tip loading area, a PCR reagent storing and mixing processing area, and a waste area.
  16. 16 . An in vitro diagnosis method, comprising using the device for extracting and detecting nucleic acids according to claim 1 to perform nucleic acid extraction and amplification on the sample.
  17. 17 . The method of claim 16 , wherein the diagnosis is for infection source identification, genetic disease, cancer detection, or genetic mutation detection.
  18. 18 . An automated molecular in vitro diagnostic instrument, comprising the device for extracting and detecting nucleic acids of claim 1 , and a computer-based automatic control system.
  19. 19 . The diagnostic instrument of claim 18 , wherein the diagnostic instrument is configured for infection source identification, genetic disease detection, cancer detection, or gene mutation detection.
  20. 20 . The diagnostic instrument of claim 19 , wherein the diagnostic instrument is configured for detection of a pathogen or a cancer, wherein the pathogen is selected from the group consisting of influenza virus, enterovirus, hepatitis B virus, hepatitis C virus, Ebola virus, Marburg virus, SARS virus, Zika virus, Bunya virus, rhinovirus, Mycobacterium tuberculosis, Escherichia coli, Acinetobacter baumannii, Streptococcus pneumoniae, Streptococcus lactis, Urea sporococcus, Staphylococcus aureus, Bacillus subtilis, Bacillus anthracis, Streptococcus, Proteus, Vibrio cholerae , and Treponema pallidum , or wherein the cancer is selected from the group consisting of gastric cancer, liver cancer, lung cancer, esophageal cancer, cervical cancer, breast cancer, colon cancer, rectal cancer, nasopharyngeal cancer, ovarian cancer, kidney cancer, bladder cancer, thyroid cancer, skin cancer, malignant tumors that grow from mesenchymal tissues including muscle, fat, bone, blood vessels, lymph, rhabdomyosarcoma, leiomyosarcoma, fibrosarcoma, liposarcoma, osteosarcoma, chondrosarcoma, angiosarcoma, lymphosarcoma, leukemia, Hodgkin's disease, Wilm's tumor (Wilms tumor), melanoma, retinocytoma, seminoma, granulosa cell tumor, Krukenberg tumor, Ewing's tumor, malignant hemangioendothelioma, and Paget's disease of the breast.

Description

This application is the U.S. national phase of International Patent Application No. PCT/CN2020/113968, filed on Sep. 8, 2020, which claims the priority of the following Chinese patent applications: the invention titled “instrument and method for extraction and detection of nucleic acids” with application No. 201910853057.9 filed on Sep. 10, 2019; and the invention titled “instrument and method for extraction and detection of nucleic acids” with application No. 201911403549.4 filed on Dec. 31, 2019”, which are incorporated herein by reference in their entirety. TECHNICAL FIELD The present invention relates to the field of in vitro diagnostic devices. Specifically, the present invention provides an automated instrument for extraction and amplification of nucleic acids, and a method for detection and diagnosis using the instrument. BACKGROUND OF ART The extraction of biologically active substances (such as cells, proteins, nucleic acids and other biologically active substances) plays an important role in modern clinical disease diagnosis, blood transfusion safety, forensic identification, environmental microbial testing, food safety testing, molecular biology research and other fields. Among them, nucleic acids are biologically active substance compounds formed by the polymerization of many nucleotides, which is one of the most basic substances of life. Nucleic acids are widely found in all animals, plant cells, microorganisms, and living organisms. According to the chemical composition, nucleic acids can be divided into ribonucleic acid, referred to as RNA and deoxyribonucleic acid, referred to as DNA. DNA is the main material basis for storing, replicating and transmitting genetic information, and RNA plays an important role in the process of protein synthesis. With the rapid development of biotechnology, with the application of PCR technology in various fields, including medical disease detection, agricultural GMO detection and many other applications, methods and instruments for extracting nucleic acid from samples and detecting them are required. However, at present, regardless of the methods, high-throughput fully automatic instruments have become the bottleneck in the application of PCR technology in the field of diagnostic analysis. There are several reasons for this. First, many diagnostic analyses are performed only with highly specialized equipment, which is expensive and can only be performed by trained clinicians. There is often only one such device in certain regions. This means that most hospitals need to send samples to these locations for analysis, thus incurring shipping costs and delays, and may even result in sample loss or incorrect functioning. Second, the equipment in question generally does not run as needed, but runs in batches after accumulating a certain amount, so there is a delay in processing for many samples, which creates a lot of uncertainty about the accuracy of the final result influence. Therefore, there is a need for instruments and methods for extraction and analysis of nucleic acid that are as automated as possible, while being suitable for operation at any time, and better able to avoid contamination. For example, once extracted from a patient, the biological samples must be placed in a form suitable for PCR processing to amplify the target vector such as nucleotide. Once amplified, the target nucleotide from the sample needs to be clearly identified. Also, the instrument should have high throughput, especially for a single sample to perform multiple different analyses; and in a manner that can be routinely performed at the point of care, without the need to send the sample to specialized equipment for testing. SUMMARY OF THE INVENTION The device for extracting and detecting nucleic acids and the automated molecular in vitro diagnostic instrument provided by the present invention have high throughput, and can flexibly allocate throughput according to requirements, and can not only detect multiple samples at the same time, but also conduct a variety of different tests and analysis on one sample. Specifically, the present invention provides a device for extracting and detecting nucleic acids, the device comprises: one or more sample receiving modules, each sample receiving module comprises a chamber adapted to receive a sample or sample container;one or more lysing modules, each lysing module comprises a chamber adapted to receive a lysing reagent or a lysing kit;one or more extraction module of nucleic acids, each extraction module of nucleic acids comprises a chamber adapted to receive nucleic acid extraction reagents or nucleic acid extraction kits;one or more amplification module of nucleic acids, each amplification module of nucleic acids comprises a plurality of chambers adapted to receive nucleic acid amplification reagents or nucleic acid amplification kits; anda detection module. In one aspect of the present invention, the sample receiving module in the device has