Automatic real-time PCR system for the various analysis of biological sample
Inventors
Park, Han Oh • Kim, Kwon Sic • Lee, Yang Won • Lee, Jin Il • Jeong, Byung Rae • Kim, Jong Hoon
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Assignees
Member
BioneerBioneer is a biotechnology company specializing in molecular biology, offering solutions in nucleic acid synthesis, molecular diagnostics, protein production, and laboratory automation. The company's expertise covers oligonucleotide chemistry, gene synthesis, sample preparation, molecular diagnostics, and analytical services such as mass spectrometry and sequencing, serving clinical, research, and industrial sectors globally.
Bioneer is a biotechnology company specializing in molecular biology, offering solutions in nucleic acid synthesis, molecular diagnostics, protein production, and laboratory automation. The company's expertise covers oligonucleotide chemistry, gene synthesis, sample preparation, molecular diagnostics, and analytical services such as mass spectrometry and sequencing, serving clinical, research, and industrial sectors globally.
Publication Number
US-9850530-B2
Publication Date
2017-12-26
Expiration Date
Abstract
The present invention relates to an automatic real-time quantitative amplification system which can perform analysis of various biological samples, and more particularly to an automatic real-time quantitative amplification system in which a plurality of decks for respectively accommodating biological samples are put in a deck storing/transferring device, whereby it is possible to automatically analyze an amount or existence of a target substance containing a target nucleic acid in the biologic sample, such as a particular gene, a particular, a particular pathogenic bacterium and a particular protein, by amplifying the target nucleic acid purified by some processes of purification, purification after culture, or purification after reaction of the target substance contained in the biological sample and then checking an amount of the amplified target nucleic acid.
Core Innovation
The invention describes an automatic, real-time quantitative PCR platform for multi-purpose biological analysis. The system includes a deck for loading a multi-well plate for treating the biological sample, a multi-well plate for PCR in which a reaction mixture for real-time quantitative PCR is injected, and an automatic purification and reaction preparing device that automatically purifies target nucleic acid and dispenses the purified target nucleic acid to the PCR multi-well plate.
The system includes a temperature-controlled deck storing and transferring mechanism that loads and transports multiple decks holding different multi-well plates, including a biological sample plate, purification-related plates for nucleic-acid purification, and a PCR reagent plate. An automatic purification and reaction preparation device includes a syringe-based block and purification and reaction-preparation pipettes that move among the biological sample plate and the purification and PCR reagent plates, and applies magnetic fields using liftable rod-shaped magnets for nucleic-acid capture, cleaning, and elution.
Purified target nucleic acids or purified binding target nucleic acids produced by antigen-antibody reaction with a target antigen contained in the biological sample are dispensed into a PCR multi-well plate and mixed with a reagent for real-time quantitative PCR. The sealed plate is transferred to a centrifugal separator, optionally mixed by a vortex mixer, and then moved to a real-time amplification device for quantitative readout. The automatic system further includes a sealing device, a centrifugal separator, and a heating unit with a liftable heating block.
Claims Coverage
The independent claims cover an automatic real-time quantitative amplification system with automatic purification and reaction preparing, deck handling, sealing, centrifugation-based residue movement, and real-time quantitative amplification. Across the provided material, independent claim coverage includes at least five main inventive feature sets.
Automatic purification and reaction preparation for real-time quantitative amplification
An automatic purification and reaction preparing device that automatically purifies target nucleic acid or cultured target nucleic acid from the biological sample and dispenses the purified target nucleic acid or the cultured and purified target nucleic acid to the multi-well plate for PCR, and mixes the dispensed target nucleic acid with a reagent for real-time quantitative PCR; or that automatically purifies binding target nucleic acid bound with a target antigen contained in the biological sample by an antigen-antibody reaction and dispenses the purified binding target nucleic acid to the multi-well plate for PCR and mixes the dispensed binding target nucleic acid with the reagent for real-time quantitative PCR.
Automatic deck storing and moving device for transferring PCR workflow
An automatic deck storing and moving device including a storing case with a door for taking in or out the deck and an internal portion maintained at a predetermined temperature, a stacking rack lifting unit moving a stacking unit up and down with racks arranged to receive the deck, and a deck transferring unit movable in a left and right direction transferring the deck loaded in the stacking rack to the automatic purification and reaction preparing device.
Sealing and centrifugation residue transfer to well bottoms
A sealing device that seals an upper surface of the multi-well plate for PCR in which the purified target nucleic acid, the cultured and purified target nucleic acid, or the purified binding target nucleic acid is dispensed, and a centrifugal separator that applies centripetal force to the multi-well plate for PCR so that a substance remained on a side wall of each well is separated and moved to a bottom surface of each well.
Syringe block dispensing architecture
The automatic purification and reaction preparing device comprises a syringe block formed with a plurality of first mounting portions to which a plurality of pipettes are removably mounted, and a syringe block moving unit that moves the syringe block so the pipettes mounted in the plurality of first mounting portions are located just above each of the multi-well plate for treating the biological sample and the multi-well plate for PCR.
Vertically liftable magnet assembly with rod-shaped magnets enclosing wells
A magnetic field applying unit in which a magnet assembly is vertically liftable and comprises rod-shaped magnets arranged so that the upper portion of the magnet encloses each well formed in the multi-well plate for magnetic particle dispersion solution when the magnet mounting block is lifted.
Liftable heating block with mounting grooves
A heating unit comprising a liftable heating block with mounting grooves for uniform heating of wells in the multi-well plate.
Sealing film mechanism with compressing, cutting, and thermal compression
A sealing device that includes a lower compressing portion and an upper compressing portion to compress a sealing film, a film cutter moved down to cut the compressed film, and a film heating block that thermally compresses the sealing film onto the PCR multi-well plate.
Vortex mixing before centrifugation
A vortex mixer that vibrates the PCR multi-well plate after it is moved from the sealing device by the moving device and before being transferred to the centrifugal separator, to mix the substance injected into the PCR multi-well plate.
The provided claim material centers on an integrated automatic workflow from nucleic-acid purification through automated syringe-block dispensing, sealing, centrifugation-based movement of substances from well side walls to well bottoms, and real-time quantitative amplification, with specific mechanical subsystems including a vertically liftable rod-magnet assembly, a liftable heating block, a sealing film compression/cutting/thermal compression mechanism, and vortex mixing prior to centrifugation.
Stated Advantages
High-throughput processing with reduced manual labor and automation from purification to PCR readout.
Rapid microorganism-related analysis including viable-cell measurement and antibiotic susceptibility testing with subsequent quantitative or relative qPCR readout.
Quantitative Immuno-PCR enabling antigen/protein quantification.
Documented Applications
Analysis of various biological samples using an automatic real-time quantitative amplification system.
Immuno-PCR antigen-density determination using purified binding target nucleic acid produced by antigen-antibody reaction.
Viable-cell measurement with culture for less than 5 generations and relative quantification.
Antibiotics susceptibility testing using parallel antibiotic conditions followed by subsequent relative qPCR.
Quantitative Immuno-PCR for small proteins/antigens using antigen-antibody reactions and quantitative readout.
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