WO2011037348A2 - Système d'analyse de micro-organisme, et procédé d'analyse l'utilisant - Google Patents

Système d'analyse de micro-organisme, et procédé d'analyse l'utilisant Download PDF

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Publication number
WO2011037348A2
WO2011037348A2 PCT/KR2010/006266 KR2010006266W WO2011037348A2 WO 2011037348 A2 WO2011037348 A2 WO 2011037348A2 KR 2010006266 W KR2010006266 W KR 2010006266W WO 2011037348 A2 WO2011037348 A2 WO 2011037348A2
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WIPO (PCT)
Prior art keywords
chamber
sample
medium
analysis device
microbial analysis
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PCT/KR2010/006266
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English (en)
Korean (ko)
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WO2011037348A3 (fr
Inventor
유재천
Original Assignee
일렉트론 바이오 주식회사
성균관대학교 산학협력단
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Publication of WO2011037348A2 publication Critical patent/WO2011037348A2/fr
Publication of WO2011037348A3 publication Critical patent/WO2011037348A3/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
    • G01N2333/24—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
    • G01N2333/255—Salmonella (G)

Definitions

  • a chromogenic medium, agar medium, and selective medium having a specific color according to the culture state of a particular bacterium by the biochemical reaction of a sugar in a medium and an enzyme in a microorganism.
  • the present invention relates to a microbial analysis apparatus capable of detecting microorganisms present in food, milk, dairy products, agricultural products, beverages, and environmental samples using any one or more of the selective medium, and an analysis method using the same.
  • the present invention relates to a microbial analysis apparatus capable of counting colonies and an analysis method using the same.
  • the cultures are differentiated by seeing the color of the grown colonies.
  • the microorganism analyzing apparatus of the present invention and an analysis method using the same are suitable for a thin film type apparatus for diagnosing and detecting a small amount of microorganisms present in a sample for contaminants such as food poisoning test, food microbial test, and water quality.
  • the existing microbial analysis process has a problem that not only the data may be wrong due to microbial contamination during these processes due to manual operations such as platinum sterilization and streaking inoculation, but only a highly expert expert can perform this task. .
  • the present invention has been made to solve the above-described problems, the object of the present invention, a sample injection port for injecting a sample; A sampler for collecting a sample from an inspection site and injecting the collected sample into the sample inlet; A prep chamber for temporarily storing a sample injected from the sample inlet; An enrichment chamber which takes a sample from the prep chamber and includes a liquid culture medium for enriching the microorganisms present in the sample; At least one medium chamber for providing nutrients to the enriched microorganism; A trech chamber for collecting debris and excess sample; And a valve for moving fluid and sample between the chambers; A flow path (channel) for providing a movement path of the specimen; And it provides a microbial analysis device having a rotatable body in which the flow path, valve and chamber are integrated and an analysis method using the same.
  • a sample injection port for injecting a sample sample;
  • a sampler for collecting a sample from a test site and including a liquid culture solution for enriching microorganisms present in the sample and injecting the sample into the sample inlet;
  • a prep chamber for temporarily storing a sample injected from the sampler;
  • a medium chamber for taking a sample from the prep chamber and providing nutrients to the microorganisms in the sample;
  • a trech chamber for collecting debris and excess sample;
  • a valve for moving fluid and sample between the chambers;
  • a flow path (channel) for providing a movement path of the specimen;
  • it provides a microbial analysis device having a rotatable body in which the flow path, valve and chamber are integrated and an analysis method using the same.
  • another object of the present invention (a) injecting a sample to the prep chamber using a sampler; (b) transferring the sample in the preparation chamber to the media chamber; (c) applying and inoculating the medium surface in the medium chamber with the sample transferred to the medium chamber; (e) It provides an analysis method using a microbial analysis apparatus according to the present invention comprising a cleaning step of collecting the surplus specimen in the trech chamber after application inoculation.
  • the present invention is a sample injection port for injecting a sample;
  • a sampler for collecting a sample from an inspection site and injecting the collected sample into the sample inlet;
  • a prep chamber for temporarily storing a sample injected from the sample inlet;
  • An enrichment chamber containing a sample from the prep chamber and containing a liquid culture medium for enriching the microorganisms present in the sample;
  • At least one medium chamber for providing nutrients to the enriched microorganism;
  • a trech chamber for collecting debris and excess sample;
  • a valve for moving fluid and sample between the chambers;
  • a flow path (channel) for providing a movement path of the specimen;
  • a rotatable body in which the flow path, the valve and the chamber are integrated.
  • the body is mixed with the disk.
  • the body is composed of an upper body and a lower body and is preferably combined to form a body.
  • the sampler preferably comprises a microbial extract for extracting microorganisms from the collected samples.
  • the sampler is preferably a pipette for sampling a sample incubated with liquid enrichment medium in a sterilization pack.
  • the liquid white liquor or liquid enrichment medium is mEC Broth, RV (Rappaport Vassiliadis) Broth, Listeria Enrichment Broth, UVM-modified Listeria Enrichment Broth, Fraser Broth, Tryptone Sonya Broth (TSB), PBS (phosphate buffered saline) Preference is given to any one selected from among Peptone water, lactose broth and desoxycholate lactose broth.
  • the medium chamber is XLD Agar, MacConkey Agar, Desoxycholate, Citrate Agar, Bismuth Sulfite Agar, Nutrient Agar, TSI Agar, Sorbitol MacConkey Agar, Oxford Agar, PALCAM Agar, LPM Agar, Mannitol Salt Agar, Baird-Parker Agar, MYP Agar, It is preferred to include at least one selected from chromogenic media and solid media.
  • sample inlet for injecting a sample sample
  • a sampler for collecting the sample from the test site and including a liquid culture solution for enriching the microorganisms present in the sample and injecting the sample into the sample inlet
  • a prep chamber for temporarily storing a sample injected from the sampler
  • a medium chamber for taking a sample from the prep chamber and providing nutrients to the microorganisms in the sample
  • a trech chamber for collecting debris and excess sample
  • a valve for moving fluid and sample between the chambers
  • a flow path (channel) for providing a movement path of the specimen
  • a rotatable body in which the flow path, the valve and the chamber are integrated.
  • the body of the present invention may further include a cover means capable of opening and closing allowing identification experiment.
  • This cover means allows the collection of a sample of suspected colonies in the media chamber.
  • identification experiment refers to an experiment in which bacteria are cultured through a medium, samples of suspected colonies are sampled, and confirmed through biochemical, serological and genetic experimental methods.
  • the body of the present invention may further include an oxygen supply valve for supplying oxygen to the aerobic microorganisms.
  • the oxygen supply valve is closed during the distribution period, and is opened at the start of the culture, thereby preventing the drying of the medium in the media chamber during the distribution period and supplying oxygen to the aerobic microorganisms during the culture.
  • the solid medium contains a large amount of water. Therefore, if cultured straight, evaporation can occur, and the upper body ceiling may be moist and humid, and the culture state of the microorganisms may be damaged, and the medium may dry out immediately during the culture.
  • the medium in the medium chamber of the present invention is preferably a solid medium, in which case the medium is installed on the body side.
  • the microorganism analyzing apparatus of the present invention preferably includes a microscopic sensor for observing microorganisms and a central control unit for processing images from the microscopic sensors and counting colonies.
  • Microbial culture usually requires a culture time of 24 to 48 hours. However, by viewing the microorganisms in the medium chamber under a microscope sensor, not only can the microorganisms grow earlier, but also the survival of the strains can be confirmed by changing the size of colonies over time. As colonies grow in size over time, the living colony can be considered a living cell.
  • the microscope sensor has an optical zoom of 20 to 100 times.
  • the sampler of the present invention may further comprise a homogenizing means.
  • the microbial analysis device is characterized in that it further comprises a temperature control device to a thermostat for providing optimum conditions of microbial enrichment.
  • the microbial extract or diluent is preferably distilled water (DeIonize water, DI water), PBS (Phosphate Buffered Saline).
  • the liquid culture solution is preferably peptone water, meat extract, yeast extract.
  • the body of the microbial analysis apparatus of the present invention is preferably a disk such as a conventional CD-ROM, DVD and the like.
  • the thickness of the body is preferably 2mm to 100mm, the diameter of 30mm ⁇ 120mm is preferred, the round disk of 120mm, 80mm, or 32mm is preferred.
  • the medium chamber is composed of a selective medium that selectively reacts with different types of microorganisms, and therefore, it is preferable to simultaneously analyze a plurality of microorganisms.
  • the medium chamber is composed of one or more medium selected from phlorogenic medium, chromogenic medium, differentiation medium, selection medium, characterized in that for analyzing a plurality of microorganisms at the same time.
  • the "selective medium” is a medium used for selectively cultivating only desired microorganisms by promoting the growth of specific microorganisms and inhibiting the growth of other microorganisms, for example, MacConkey Agar medium, Salmonella-shigella medium and the like. Can be used.
  • the "fluorogenic medium” refers to a medium that exhibits a unique fluorescent light depending on the culture state of a specific bacterium by the biochemical reaction of a sugar in a medium and an enzyme in a microorganism.
  • Microbial analysis apparatus is Salmonella (Salmonella), Bacillus (Bacillus), Listeria, Vibrio, Campylobacter S. aureus, E. Coliform, E. coli, E. coli, Shigella, Legionella Enterobacter sakazakii, Citrobacter, Preteus, Citrobacter, MRSA, E.coli O157, E.coli spp, L.monocytogenes, L.inocua, V.parahaemolyticus, V.vulnificus & V.cholerae, V.alginolyticus, Coliforms, Pseudomonas Microorganisms such as Klebsiella, Citrobacter, Enterococcus, PMP group, Staphylococcus saprophyticus, Stapylococcus aureus, S. agalactiae, Candida are preferred.
  • microorganisms have a unique color depending on the culture state of specific bacteria by colony formation, biochemical reactions of various metals in the medium, dyes or sugars in the medium and enzymes in the microorganisms. Color readings and colony counts allow quantitative analysis of the presence and presence of specific microorganisms. For example, microbes E. coli are red, Streptoccoccus is cyan, Proteus is brown, E. coli O157 is soft purple, and Coliform is blue.
  • the valve attaches a thin film adhesive tape to the pores to close the pores during the distribution storage period, thereby completely closing the hydraulic pressure formed in the fluid by the centrifugal force of the disk.
  • a valve is preferable in which the thin film adhesive tape is peeled from the pores with a force to open the pores.
  • a valve for releasing the thin film adhesive tape from the pores and opening the pores by the force of hydraulic force formed in the fluid itself by the centrifugal force of the disk is referred to as a "hydraulic burst valve”.
  • This "hydraulic burst valve” is not only a valve in the form of a thin film, but also a thin film adhesive tape (flexible), it is well adapted to expansion and contraction according to environmental factors such as temperature, so that the liquid evaporation or There is an advantage that the sealing (sealing) problem due to the expansion and contraction of the body does not occur.
  • the closing strength is determined by the adhesive area of the thin film adhesive tape when the hole is closed by the thin film adhesive tape, and the thin film adhesive tape is torn apart at a disk rotation speed (centrifugal force) that overcomes the closing strength.
  • the pores open.
  • the thin film adhesive tape is weakened by the heat of the irradiated laser beam, and the opening of the valve is desired, the thin film adhesive tape is easily detached from the pore by applying the laser beam to the hole.
  • the valve is preferably a valve for attaching a thin film adhesive tape to close the pores during the distribution storage period to completely close the open pores and cutting the thin film adhesive tape by the heat of the laser to open the pores.
  • a valve that opens the pores by cutting the thin film adhesive tape by the heat of the laser is referred to as a "laser burst valve”.
  • the color of the thin film adhesive tape is preferably black, in order to absorb the laser beam well.
  • the valve seals the pores by the adhesive force or the binding force between the pores and the stopper to block the pores during the distribution storage period, and the stopper is separated from the pores by the centrifugal force by the high-speed rotation of the disc in use. It is an object of the present invention to provide a microbial analysis device using the open stopper valve and an analysis method using the same.
  • the stopper is preferably an adhesive material which melts by heat or is coated with an adhesive on the face of the hole for adhesion with the stopper and the hole.
  • a laser beam absorbing material it is also preferred to coat a laser beam absorbing material around the stopper to absorb the laser beam.
  • a laser beam absorbing material has a property of easily generating heat by absorbing a laser beam during laser beam irradiation.
  • the pressure sensitive adhesive is weakened or melted by the heat of the irradiated laser beam. Therefore, if you want to open the valve, apply a laser beam to the stopper is easily detached by the rotational force.
  • stopper burst valve a valve in which the stopper blocking the hole by the centrifugal force of the disk overcomes the adhesive or bonding force between the stopper and the hole and is separated from the hole so as to open the hole.
  • the stopper burst valve is referred to as a "stopple burst valve”.
  • bush burst valve is preferably a bead or thin film cylindrical stopper, but the shape is not particularly limited as long as it can perform the above-described functions or functions.
  • the valve seals the oil hole by the attraction force between the constrained groove made of a magnetic material and the magnetic valve, and the magnetic valve may be released from the constrained groove by the centrifugal force by the rotation of the disk to open the oil hole.
  • the magnetic valve which blocks the hole by the centrifugal force of the disk overcomes the attraction force with the restraint groove and is separated from the restraint groove to open the hole. It is called).
  • magnetic burst valve is preferably a bead, a thin film cylindrical magnet or a thin film rectangular magnet, but the shape is not particularly limited as long as it can perform the above-described functions or functions.
  • the magnet valve that has been removed from the restraint groove is closed again by the attraction force with the restraint groove.
  • the magnet valve is preferably demagnetized by the heat of the laser beam by irradiating the laser beam, so that the magnetic force is weakened. Therefore, if you want to open the valve and apply a laser beam to the magnet valve, the magnet valve is easily opened by the rotational force.
  • the invention is characterized in that the valve is made by a hydrophobic channel formed between the upper body and the lower body. Since the hydrophobic channel is hydrophobic, it is difficult for the fluid to move normally and the fluid can pass only through the centrifugal force generated when the body rotates.
  • the hydrophobic channel is formed between the upper body and the lower body when the upper body and the lower body are bonded by a thin film adhesive tape having a channel shape. That is, the upper body and the lower body are closely attached to each other by a thin film adhesive tape to form a single body, wherein a hydrophobic thin film channel is formed by a portion in which the thin film adhesive tape is missing between the upper body and the lower body, that is, the channel shape. .
  • the height of the hydrophobic thin film channel is determined by the thickness of the thin film adhesive tape.
  • a valve that overcomes the resistance to hydrophobicity preventing the fluid from moving by the centrifugal force of the body and moves the hydrophobic thin film channel is referred to as a "hydrophobic burst valve”.
  • burst valve the "hydraulic burst valve”, the “bump burst valve”, the “magnetic burst valve, the” laser burst valve "and the” hydrophobic burst valve "will be collectively referred to as” burst valve ".
  • the burst valve is opened by heat generated by the laser beam and centrifugal force or centrifugal force.
  • the laser beam from the laser beam generator is irradiated around the burst valve to weaken the sealing force or coupling force of the burst valve by heat by the laser beam so that it can be easily opened by centrifugal force.
  • the laser beam generator is preferably mounted on a slider.
  • the first method is to move the laser beam generator by moving the laser beam to a radial position of the burst valve in order to irradiate the laser beam to the burst valve during disk rotation.
  • a laser beam is irradiated to the burst valve, in which case the amount of laser beam irradiated to the burst valve is the rotational speed of the disc and the laser
  • the laser beam irradiation operation during the rotation of the disk which is a function of the power of the beam generator, is hereinafter referred to as " scanning beam " operation in the present invention.
  • the laser beam generator is moved by moving the laser beam to the burst valve at the radial position of the burst valve during disk rotation, and the laser beam generator and the corresponding burst valve are rotated during the disk rotation.
  • the laser beam is turned on every time (ie, encountering) and off otherwise.
  • This laser beam irradiation operation is hereinafter referred to as "pulse beam" operation in the present invention.
  • the opening of the burst valve is characterized by disk rotation after heating of the burst valve by the "pulse beam” operation or the “scanning beam” operation.
  • the opening of the burst valve on the concentric circle of the microbial analyzer is performed by the centrifugal force generated in the fluid during the rotation of the microbial analyzer and the laser beam generator installed on the slider during the rotation of the microbial analyzer and the burst on the concentric circle.
  • a "pulse beam” or “scanning beam” operation in which the burst valve is heated to open each time it matches a valve, such a “pulse beam” or “scanning beam” operation causes the high viscosity fluid to move.
  • Fluid with high viscosity is easy to move with centrifugal force. Highly viscous fluids may not move to neighboring chambers even when the valve is open during shutdown, which may cause reproducibility and reliability problems.
  • the disk preferably has at least one medium chamber arranged concentrically.
  • the image sensor for photographing the medium chamber of the disk; And a spindle motor for rotating the disk.
  • the entire shooting of the concentric discharge chamber may be performed by an image sensor while rotating the spindle motor 360 degrees slowly, or by repeating the rotation and stopping of the spindle motor repeatedly to obtain an image at each stop and combine them. Get a 360 degree full image.
  • the image sensor is characterized in that the microbial colonies (colony) in the medium chamber by color classification and counting and quantitative analysis.
  • the count is a measure of the number of bacteria forming colonies in the medium chamber, and plate count or colony count is preferred.
  • the coefficients are expressed in bacterial counts or colony-forming units (cfu / ml or cfu / g).
  • the image sensor is characterized in that reading a barcode (barcode) on the disk.
  • the image sensor reads a bar code on the disc to authenticate the authenticity of the disc, or the data of the bar code and the user's information (for example, IP address, location, telephone number, e_mail address, company name, or The serial number of the disk, etc.) is remotely transmitted through the Internet to receive notification of authenticity of the disk and product identification information from the server.
  • the user's information for example, IP address, location, telephone number, e_mail address, company name, or The serial number of the disk, etc.
  • the microorganism analyzing apparatus may be connected to a computer or an internet network by the input / output device.
  • the microbial analyzer cannot drive the disc.
  • the disc is genuine, the barcode is registered in the central server, and when the product identification information is received from the central server, the microbial analysis apparatus drives the disc according to the driving software corresponding to the product identification information. If the driving software corresponding to the product identification information is not installed in the computer or the microbial analysis apparatus, the driving software is downloaded from the central server through the Internet network.
  • the storage device or the RF IC stores the usage history and the analysis result of the disk.
  • the central control unit is characterized in that the information stored in the storage device or RF IC to read and transmit to the server remotely through the input and output device.
  • a warning message is sent to the user, whether to be automatically ejected when loading a disc which is not usable in a conventional optical disc (for example, CD, DVD) or an unrecognized microbial analysis device.
  • a conventional optical disc for example, CD, DVD
  • an unrecognized microbial analysis device for example, CD, DVD
  • the present invention (a) injecting a sample to the prep chamber using a sampler; (b) transferring the sample in the preparation chamber to the media chamber; (c) applying and inoculating the medium surface in the medium chamber with the sample transferred to the medium chamber; (e) It provides an analysis method using a microbial analysis apparatus according to the present invention comprising a washing step of collecting the surplus specimen in the trech chamber after application inoculation.
  • the analysis method may further comprise a enrichment culture step.
  • the analysis method may further include reading the medium chamber by the image sensor.
  • the analysis method may further comprise a medium chamber reading step and the microbial colonization step.
  • the analysis method may further include transmitting the medium chamber reading result to a remote server.
  • the present invention provides a sample injection port for injecting a sample;
  • a sampler for collecting a sample from an inspection site and injecting the collected sample into the sample inlet;
  • a prep chamber for temporarily storing a sample injected from the sample inlet;
  • An enrichment chamber containing a sample from the prep chamber and containing a liquid culture medium for enriching the microorganisms present in the sample;
  • At least one medium chamber providing a culture medium for providing nutrients to the enriched microorganisms;
  • a trech chamber for collecting debris and excess sample;
  • a valve for moving fluid and sample between the chambers;
  • a flow path (channel) for providing a movement path of the specimen;
  • it provides a microorganism analysis device having a rotatable disk-shaped body in which the flow path, valve and chamber are integrated and an analysis method using the same.
  • the body may be selected from various materials such as plastic, glass, mica, silica, silicon wafer, and the like.
  • plastics are preferred for economic reasons, ease of processing, and compatibility with existing laser reflection based detectors such as CD-ROM and DVD readers.
  • Plastics that can be used include polypropylene, polyacrylates, polyvinyl alcohol, polyethylene, polymethyl methacrylate (PMMA), Cyclic Olefin Copolymer (COC), acrylics and polycarbonates. Of these, polypropylene, COC, acrylic and polycarbonate are preferred, and acrylic, COC and polycarbonate are most preferred.
  • the surface of the body may also be aluminum coated to prevent evaporation of the liquid stored in the chamber.
  • the prep chamber, enrichment chamber, medium chamber, trech chamber; And a flow path, a hole, and a valve for fluid movement between the chambers are formed in the disc-shaped body to constitute various processes required for microbial analysis, and the disc-shaped body is formed by stacking an upper body and a lower body. Characterized in that configured.
  • the cover means is closed by a magnetic force forming an attractive force between the upper body and the lower body, and the experimenter collects a sample of the suspected colony from the medium chamber through the opening of the cover means for identification experiments. It is done.
  • the cover means is preferably a permanent magnet is installed in the upper body and a ferromagnetic material is installed in the lower body is closed by the attraction between the upper body and the lower body.
  • cover means further comprises an opening means, allowing easy opening by the opening means.
  • the opening means is preferably an electromagnet board in which an electromagnet is generated which generates a repulsive force on the permanent magnet in the upper body.
  • an electromagnet board on (ON) When the electromagnet board on (ON) generates a repulsive force for the permanent magnet in the lower body, it is possible to easily separate the lower body from the body.
  • the upper body and the lower body may be combined by ultrasonic welding or laser welding, a thin film adhesive tape. Since such lasers generate heat well at the interface between materials having different light transmittances, it is preferable that the upper body and the lower body use different kinds of plastic materials for laser welding.
  • the thin adhesive tape is preferably an adhesive (a gluing agent) used for all adhesive tapes such as double-sided tape, and the adhesive is silicone, rubber, modified silicone, acrylic, Materials such as polyester, epoxy and the like can be used.
  • the double-sided tape is surface treated with an adhesive (a adhesive) on both or one side of the release paper such as paper, vinyl, polyester film, polyethylene film and other synthetic materials.
  • an adhesive a adhesive
  • the release paper such as paper, vinyl, polyester film, polyethylene film and other synthetic materials.
  • a pressure-sensitive adhesive material having characteristics such as high sealing and buffering, vibration relaxation, impact resistance, heat resistance, adsorption, and adhesive strength.
  • a thin film coating by adhesive on one side of the body or by applying a dispenser or spray or silk screen printing on the adhesive It is preferable to apply a thin film coating on one side of the adhesive.
  • the body of one microorganism analyzing apparatus is assembled by attaching a thin film-coated upper body and a lower body to each other in close contact with each other.
  • a pore closure membrane of the double-sided tape "hydraulic burst valve” is included, so that it is preferred to leave a pore-closing membrane with an adhesive on the pore area when removing the release paper or to form a pore-closing membrane for the "hydraulic burst valve" during the thin film coating.
  • the porous closed membrane may be a membrane that can be easily torn by hydraulic or centrifugal force instead of a thin film adhesive tape, and the surface of the membrane is hydrophobic without affinity with water.
  • Polymer membranes such as polyethylene (PE), polypropylene (PP), polysulfone (PS), polyalkene, cellulosics, polyvinyl, polycarbonate, and polyamide may be used as the membrane.
  • the hydrophobic membrane surface effectively blocks the movement of fluid stored in the chamber.
  • the stopper is characterized in that the cylinder, or a ball (ball) or a thin film selected from the shape of the cylinder.
  • the stopper is a pressure-sensitive adhesive coated wax or a wax is preferred.
  • the wax is preferably paraffin wax, synthetic wax, or microcrystalline wax.
  • the adhesive or the adhesive coated on the surface of the plug is preferably melted or weakened by the heat of the irradiated laser beam.
  • the adhesive has a weak adhesive strength of the burst valve due to the heat of the irradiated laser beam, and the burst valve is opened by the disk rotation.
  • Microbial analysis device and an analysis method using the same of the present invention is suitable for detecting a small amount of microorganisms in the fluid.
  • the microbial analysis apparatus according to the present invention and an analysis method using the same provide an effect of automating a series of processes such as microbial enrichment, application (inoculation), culture, and detection.
  • 1 to 4 is a view showing various embodiments of a burst valve used in the microbial analysis apparatus according to an embodiment of the present invention
  • 5 to 7 illustrate various embodiments of a “magnetic burst valve” using a cross sectional view of the “magnetic burst valve” and a magnet valve embedded in a disc;
  • FIG. 8 to 10 is a view schematically showing a microbial analysis apparatus according to an embodiment of the present invention.
  • FIG. 11 is another embodiment of a turntable and a crimping means
  • FIG. 14 is a view showing a sampler used in the microbial analysis apparatus according to an embodiment of the present invention.
  • 15 is a view showing the results of culturing microorganisms in the medium chamber
  • FIG. 16 is a diagram illustrating an embodiment of a top loading microorganism analysis device incorporating a thermostat or a temperature control device.
  • FIG. 17 is a view showing an embodiment of a front loading type microbial analysis apparatus in which a thermostat or a temperature control device is built into a tray.
  • FIG. 18 is a view showing an embodiment in which the cover means is installed on the outermost and innermost circumference of the body
  • 19 is a view showing an embodiment of an opening means using an electromagnet board
  • FIG. 20 is a view showing an embodiment of an oxygen supply valve for supplying oxygen to the aerobic microorganisms in the medium chamber
  • FIG. 21 is a diagram illustrating an embodiment of a microbial analysis apparatus using a test strip.
  • FIG. 1 to 4 illustrate various embodiments of a burst valve used in a microbial analysis apparatus according to an embodiment of the present invention.
  • FIG. 1 and 2 are cross-sectional views and exploded views of a-b of the hydraulic burst valve using a thin film adhesive tape installed in the disk or body of the microbial analyzer.
  • FIG. 2 shows that the hole closing film 13a is torn off by the hydraulic pressure generated in the liquid (not shown) by the rotation of the body 100 so that the hole 10b is opened in the chamber 11a. It indicates that the stored liquid has been moved to the chamber 11b.
  • the body 100 includes an upper body 1 and a lower body 2, each of which includes a flow path 16a through which fluid can flow on the body surface during the injection molding process; Chambers 11a and 11b capable of storing a sample or a solution such as a liquid culture solution; And a hole 10b connecting the chambers 11a and 11b.
  • the upper and lower bodies 1 and 2 are closely attached to each other by the thin film adhesive tape 2a to form one body 100.
  • the upper body 1 and the lower body 2 constitute the chambers 11a and 11b
  • the lower body 2 has a flow path 16a for connecting the chamber 11a and the chamber 11b. Is engraved to a certain depth, and a hole 10b is formed at the distal end of the flow path 16a to provide a connection between the chamber 11a and the chamber 11b. This hole 10b is closed by the hole blocking membrane 13a.
  • the hole closing membrane 13a is formed at the portion of the hole 10b by the thin film adhesive tape 2a when the upper body 1 and the lower body 2 are attached and assembled.
  • Closing the pores 10b by the hole closing membrane 13a completely blocks the pores 10b during the distribution storage period, and the fluid itself stored in the chamber 11a by centrifugal force by the high speed rotation of the body 100 during use.
  • the hole closed membrane 13a is torn off by the hydraulic pressure formed in the hole, and the hole 10b is opened to move the fluid into the chamber 11b. Since the pore-closing membrane 13b is flexible, it adapts well to expansion and contraction according to environmental factors such as temperature, and thus sealing problems due to evaporation of liquid or expansion and contraction of the body during distribution. There is an advantage that does not occur.
  • the pore-closing membrane 13b is formed in the hole 10b when the upper body 1 and the lower body 2 are attached and assembled in close contact with each other by a thin film adhesive tape 2a. It features.
  • the closing strength when closing the pores by the thin film adhesive tape 2a is proportional to the adhesion area 14 between the pore-closing membrane 13a and the lower body 2, so that the body
  • a plurality of "hydraulic burst valves" are installed in the 100 and their closing strengths are different from each other so as to open a desired valve at a desired point in time so as to provide a hydraulic pressure greater than the closing strength of the closed hole membrane 13b of the corresponding valve.
  • Hydraulic force, hydraulic pressure) to generate a centrifugal force is characterized in that the hole (10b) to be opened selectively or individually.
  • the adhesive strength of the "hydraulic burst valve” is weakened by applying heat to the hole-closing membrane 13b using a laser beam, and the hydraulic burst valve is easily opened by disk rotation.
  • the pore-closing membrane 13a is characterized by constituting a thin film adhesive tape layer by a phase change material.
  • the phase change material preferably has a melting point of 40 degrees to 70 degrees.
  • the pore-closing membrane 13a is made of a thermoplastic adhesive material, and the remaining bonding portion constitutes a thin film adhesive tape layer by a thermosetting adhesive material.
  • the pore-closing film 13a may be a thin film adhesive tape formed by a thermoplastic hot malt adhesive, and the remaining joint may be a thin film adhesive tape formed by an acrylic adhesive which is thermosetting.
  • the adhesive force of the pore-closing membrane is weakened more easily by the heat of the laser beam than in the remaining portions.
  • the pore-closing film 13a is easily softened by the heat of the ray point beam, and thus weakens the adhesive force, while the surrounding and the remaining parts are bonded by the heat-curable tape, thereby providing the advantage that the adhesive force is not weakened by the heat of the laser beam. .
  • the softening temperature of the thermosetting pressure-sensitive adhesive (Softening Temperature) is preferably 120 degrees or more, and the thermoplastic pressure-sensitive adhesive The softening point is more preferably between 60 and 80 degrees.
  • thermoplastic resin is preferably COC, PMMA, PC, PS, POM, PFA, PVC, PP, PET, PEEK, PA, PSU and PVDF.
  • 3 and 4 show an example of a “bung burst valve” using a plug embedded in the body 100.
  • the body 100 is composed of an upper body (1) and the lower body (2), each of which has a flow path (16a) through which fluid can flow on the body surface during the injection molding process; Chambers 11a and 11b capable of storing solutions; And the hole (10b) for connecting the chamber (11a, 11b) is formed.
  • the hole (10b) is closed by a stopper (13b) buried in the hole, by closing the hole by the stopper (13b) to completely block the hole during the distribution storage period, by the high-speed rotation of the body 100 in use
  • the stopper 13b is released toward the auxiliary channel 16b so that the hole 10b is opened.
  • another aspect of the present invention is because the closing strength (closing strength) when closing the hole (10b) by the stopper (13b) is proportional to the contact area between the stopper and the flow path (16a), a plurality of "stopper" in the body 100 Burst valves "and their closing strengths are different from each other so that the perforations 10b can be selectively or independently by rotating the disc to generate a centrifugal force above the closing strength of that valve to open the desired valve at a desired point in time. It is characterized in that the opening.
  • the diameter of the stopper is preferably 1mm to 5mm. As the diameter increases, the contact surface increases, thereby increasing the reliability of opening and closing.
  • the plug may be spherical but thin film particles may also be used. In the present invention, a thin film cylinder or a thin film square is preferred as the thin film particles. The thickness of the thin film particles is preferably about 0.1 mm to 2 mm.
  • FIG. 4 shows a case in which the oil hole 10b is blocked by the stopper 13b and the flow path 16a is blocked.
  • the oil hole 10b is generated by the rotation of the body 100.
  • 5-7 are various embodiments of a "magnetic burst valve” using a cross sectional view between a-b of the "magnetic burst valve” and a magnet valve embedded in the body 100.
  • Reference numeral 90 denotes a magnet valve composed of permanent magnets, and the magnet valve closes the oil hole 10b by the attraction groove and the attraction force 101 formed of the magnetic material.
  • the magnetic valve 90 is separated from the restraint groove 101 by the centrifugal force to open the oil hole 10b.
  • the boundary membrane 91 is installed so that the detached magnet valve 90 does not deviate more than a certain range.
  • the restraint groove 101 serves to prevent the magnetic valve 90 from falling off by shaking the body 100.
  • the diameter of the constrained groove 101 is preferably 20% to 70% larger than the diameter of the magnet valve 90.
  • the magnetic valve 90 may be formed of a permanent magnet and may be coated with a rubber cushion material such as silicone rubber thereon to increase the sealing force of the pores. Thin film circular magnets, thin film cylindrical magnets or thin film square magnets or ball magnets are preferred.
  • FIG. 6 shows the case where the hole 10b is opened by centrifugal force, and the figure below shows the case where the hole 10b is closed.
  • the body (1, 2) has the advantage that it does not have to be a ferromagnetic material.
  • FIG. 8 to 10 illustrate an embodiment of a disk 100 and peripheral drive devices in which various processes for microbial analysis are integrated as an embodiment of the microbial analysis apparatus 200 of the present invention. .
  • the disk 100 is composed of the upper body 1 and the lower body 2, and is adhered to each other by a thin film adhesive tape (2a) to form a single disk (100).
  • Reference numeral 120 denotes a sample injection means selected from a dispenser, pipette, dice, sampler, and lancet for injecting a sample
  • reference numeral 121 denotes a sample injection port for injecting a sample
  • Reference numeral 130 is a preparation chamber for temporarily storing a sample injected by the sample injection means 120 and performing a preparation process
  • reference numeral 132 denotes a culture medium for a culture process for providing nutrients to the microorganisms in the sample.
  • Is a media chamber Is a media chamber
  • a reference numeral 133 denotes a trash chamber for a cleaning process for collecting the residue and excess sample
  • reference numerals 70 and 71 denote a valve for moving the fluid and the sample between the chambers.
  • reference numeral 170 denotes a disk gap.
  • Reference numeral 335 denotes a reference hole indicating a reference of the body.
  • reference numeral 91 is a bar code that may include information about the product ID, expiration date, the type of microorganism that can be analyzed and diagnosed, etc. of the disc.
  • the image sensor 103 not only reads the barcode 91 on the disc, but also photographs the medium chamber 132 to quantitatively or qualitatively analyze the microorganism culture result in the medium chamber 132. Get a color image of
  • the preparation chamber 130 may further include an enrichment chamber for containing or storing a liquid culture medium for enriching the microorganisms present in the sample.
  • Reference numeral 211 denotes a slider mounted on the laser beam generator 5a and connected to a slide motor 109 to be driven and controlled to generate a laser beam for the burst valve by movement of the slider. Radial space addressing of the device 5a is achieved.
  • the radial space addressing is by means of a slide motor 109 capable of reversibly moving the slider in the radial direction.
  • the rotation of the slider motor allows the slider to move radially outward from the center of the body or from the outside of the body to the center of the body.
  • the opening and closing control of the valve at the start point and the end point of each of the above processes is performed by a burst valve.
  • the body 100 is rotated while the laser beam generator 5a installed on the slider 211 is spatially moved to the radius of the valve and the laser beam is turned on. Heating the burst valve opens the valve by centrifugal force during rotation. At this time, the laser beam generator 5a operates in the "pulse beam” or "scanning beam” mode.
  • Reference numeral 110b is a flexible cable for connecting various control signals required for the laser beam generator 5a and the image sensor 103 on the slider 211 to a wafer or a harness. 110a is connected to the central control unit 105.
  • Reference numeral 181 denotes a turn table for placing the disk 100, which is front or top loaded on the turn table through the central void 170.
  • Reference numeral 188 is a memory-embedded wireless RF IC or electronic tag device that includes protocols for analyzing microorganisms, analysis algorithms, and standard control values for reading.
  • personal encryption information and identification (identification) of the microbial analysis device can be stored, so that others can not be used without permission.
  • the wireless RF IC 188 is preferably in the form of a smart IC card.
  • the wireless RF IC 188 information is provided to the central control unit 105 through wireless transmission and reception, and is utilized for personal encryption.
  • Reference numeral 110 is a radio wave generation unit for supplying power to the wireless RF IC 188.
  • the radio wave generated by the radio wave generator generates a sufficient amount of electricity by supplying a power to the radio RF IC 188 by sensitizing the induction coil coil embedded in the radio RF IC 188 according to Fleming's law.
  • the wireless RF IC 188 has a temperature measuring function, and measures the temperature of the medium chamber to wirelessly transmit to the central control unit 105.
  • the central control unit 105 maintains a constant temperature by the heating means 240 or the cooling means.
  • the temperature of the medium chamber 132 is preferably maintained at a temperature selected between 35 degrees Celsius and 40 degrees Celsius suitable for microbial growth.
  • the cooling means is preferably by rotation or rotating fan of the body 100.
  • the disk is rotated or the fan is rotated to cool the heated medium chamber 132 by wind while cooling the medium chamber 132 to a desired temperature based on the temperature measured by the RF IC. It is preferred to rotate the disk 100 further to make it work.
  • the wireless RF IC 188 is characterized in that the temperature of the discharge chamber 132 is read and wirelessly transmits the result to the central control unit 105.
  • the heating means is preferably a nano pattern 240 connected to an output terminal of the RF IC, and the heating means 240 is controlled by on / off intervals and power amounts of power supply of the RF IC. It is preferred that the temperature of is controlled.
  • heating means is made by a scanning beam operation of the laser beam generator 5a.
  • the wireless RF IC 188 includes information on an inspection date and a test result, an expiration date of the test, and a microbial analysis result according to the test of the microbial analysis device. It is done. After the microbial analysis, the information on the microbial analysis device may be brought to the RF IC reader or the body 100 may be loaded into the microbial analysis device.
  • the microorganism analysis result is preferably a color image of the medium chamber 132 obtained by the image sensor 103.
  • the test results analyze the color image of the medium chamber 132, the colony density and population of the colony (population) according to the type of microorganism is preferred.
  • the microorganism analyzing apparatus of the present invention preferably includes a microscopic sensor 106 for observing microorganisms and a central controller 105 for processing an image from the microscopic sensor and counting colonies or checking whether cells are dead. do.
  • the central control unit 105 stores the test result, the test date, and the microbial analysis result of the disk 100 in the memory or storage device 113 embedded in the wireless RF IC 188. It is characterized by storing in).
  • the input / output device has a communication standard of USB (Universal Serial Bus) or IEEE 1394 or ATAPI or SCSI or Internet communication network.
  • USB Universal Serial Bus
  • IEEE 1394 Universal Serial Bus
  • ATAPI ATAPI
  • SCSI Internet communication network
  • information about the sample itself such as the name of the sample, the collection place of the sample, the collection date and time of the sample can be input.
  • FIG. 9 shows an embodiment of the upper view of a slider 211 in which the laser beam generator 5a and the image sensor 103 are installed.
  • the slider is movement controlled by worm gear connections 109a and 109b connected to the axis of the slide motor 109.
  • the slider is slidably moved using the slide arms 108a and 108b as guides.
  • the slide arms 108a and 108b are fastened to the body of the microbial analysis apparatus 200 through screws 110a to 110d.
  • Reference numeral 110b denotes a flexible cable and is connected through a wafer or harness 110a.
  • Reference numeral 181 denotes a turn table that is rotated by the spindle motor 102 described above.
  • reference numeral 200a denotes an outer body supporting the microorganism analyzing apparatus 200.
  • a circuit board 140 is jointly fastened to the outer body 200a at the bottom of the microorganism analyzing apparatus 200, and a central controller 105 and a storage device for controlling the microorganism analyzing apparatus 200 on the circuit board.
  • 113 and an input / output device 111 are designed to be disposed on the circuit board 140.
  • the central controller 105 not only controls the spindle motor 102 for rotation or stop of the disk 100, but 2) design arrangement on the slider 211 by the control of the slide motor 109.
  • 3) serves to move the position of the laser beam generator 5a to the radial position of the valve in order to apply heat to the valve during the valve opening and closing operation.
  • the image sensor 103 may be mounted on the slider 211 or disposed on the circuit board 140. Note that only the image sensor 103 is mounted on the slider 211.
  • the central control unit 105 determines whether the disk currently loaded in the microbial analysis apparatus 200 is for microbial analysis.
  • the unique ID of the disk 100 to the central control unit 105 via the wireless RF IC 188 at the time when the disk 100 is loaded into the microbial analysis apparatus 200.
  • the central control unit 105 recognizes whether the disk 100 currently loaded in the microbial analysis apparatus 200 is a disk for microbial analysis.
  • Another aspect of the present invention by sensing the barcode on the disk by the image sensor 103 at the time when the disk 100 is loaded into the microbial analysis device and by analyzing it by the central control device 105, The central controller 105 recognizes that the disk loaded in the analysis device 200 is a disk for analyzing microorganisms.
  • a specific mark or pattern on the disk is sensed by the image sensor 103 at the time when the disk 100 is loaded into the microbial analysis apparatus and the central control device ( By analyzing by 105, it is characterized in that the central control unit 105 recognizes whether the disk currently loaded in the microbial analysis apparatus 200 is a genuine microbial analysis disk.
  • the image information about the discharge chamber 132 obtained by the image sensor 103 may be transferred to the central control unit 105, the storage unit 113, or the input / output unit through the flexible cable 110b connected to the slider 211. 111).
  • the image information of the discharge chamber 132 obtained by the image sensor 103 on the slider 211 or the image sensor arranged and designed on the circuit board 140 may be transmitted to the central control unit 105, the storage unit 113, or the input / output unit. Is sent to the device 111.
  • Reference numeral 104 is a crimping means of the disk 100 loaded in the disk cavity 170 is preferably pressed by the magnetic force with the ferromagnetic material (104a) is preferably designed to enable vertical movement and idle rotation.
  • the ferromagnetic material (181a) is preferably compressed by the magnetic force with the turntable (181).
  • the turntable 181 and the crimping means 104 are preferably permanent magnets.
  • the slider 211 may further mount a movable permanent magnet 5b.
  • the image sensor 103 performs a "search process of the discharge chamber" before capturing an image for the discharge chamber 132.
  • the permanent magnet 5c is provided on the body 100 to facilitate optical alignment between the image sensor 103 and the medium chamber 132. It is characterized by.
  • the short rotation is preferably a rotation of 0.1 second to 0.5 seconds.
  • FIG 11 is yet another embodiment of the turntable 181, the ferromagnetic material (181a) and the pressing means 104.
  • the ferromagnetic material 181a has a groove 181b to be mechanically engaged with knobs 181c and knobs of the turntable 181.
  • Reference numeral 40 is at least one light emitting diode (LED) for illumination of the image sensor, the image sensor 103 or the LED is mounted on the slider 211 or the upper side of the discharge chamber 132 Or it can be installed at the bottom.
  • LED light emitting diode
  • the image sensor 103 is preferably a linear image sensor that senses the amount of light in a CCD or CMOS or pixel unit.
  • the linear image sensor is preferably a linear sensor array or a contact image sensor (CIS).
  • the image sensor 103 is characterized in that to move the slider 211 to the radius corresponding to the discharge chamber 132 to obtain the image information of the discharge chamber 132.
  • the disk 100 is an embodiment in which the upper body 1 and the lower body 2 are laminated and bonded by an adhesive tape 2a, and the hydraulic burst valve is employed as a valve.
  • Reference numerals 121a, 121b, 121c, and 121d indicate sample inlets for injecting a sample, and the disk 100 includes at least one prep chamber 130a to 130d for temporarily storing a sample injected through the sample inlet. ; At least one medium chamber (132a, 132b, 132c, 132d) providing a medium (80) for providing nutrients to the microorganisms in the sample; Trash chambers 133a, 133b, 133c, and 133d for collecting debris and excess samples; And valves 70a, 70b, 70c, 70d, 71a, 71b, 71c and 71d for moving the fluid and the sample between the chambers.
  • Reference numeral 170 denotes a disk gap.
  • Reference numeral 335 denotes a reference hole indicating a reference of the body.
  • Each prep chamber 130a, 130b, 130c, 130d may be injected with different or the same kind of sample.
  • the medium chambers 132a, 132b, 132c, and 132d may contain the same medium or different types of medium. Therefore, with the disk 100, it is possible to test for multiple microorganisms on the single specimen and to test a single microorganism on the multiple specimens.
  • the hydraulic burst valves 70a to 70d provided therein must be opened, respectively.
  • the internal pressures to open the hydraulic pressure valves 70a to 70d installed on the inside first.
  • the adhesion area of the thin-film adhesive tape of the burst valves 70a to 70d is larger than the hydraulic burst valves 71a to 71d provided on the outside to increase the closing strength, or the hydraulic burst valves 70a to 70d provided on the inside.
  • the hydraulic burst valves 70a to 70d installed inside by the laser beam generator 5a should be heated by a "pulse beam” or "scanning beam” operation.
  • Samples moved into the medium chamber 132 by opening the hydraulic burst valves 70a to 70d should be evenly applied on the surface of the medium 80 so that the inoculum is inoculated onto the medium.
  • the inner height 131a of the discharge chambers 132a to 132d is smaller in height than the outer height 131b. Therefore, the sample introduced into the medium chamber is collected in the direction of the center of the medium chamber by the capillary phenomenon. Subsequently, when the disk is rotated, the sample is moved outward in the radial direction of the medium chamber by centrifugal force. Then, when the disk 100 is stopped rotating, the sample is moved back inside the medium chamber by capillary action. If this is repeated several times, the sample is evenly applied to the surface of the medium (80). Thereafter, the burst valves 71a to 71d are opened to move the excess sample to the trech chambers 133a to 133d.
  • the prep chambers 130a to 130d are preferably hydrophilic coated, and the hydraulic burst valves 70a to 70d installed therein are the "magnetic burst valves", “hydrophobic burst valves” and “laser burst valves” described above. It may be replaced by any one or more of ".
  • the chamber height of the preparation chambers 130a to 130d it is preferable to design the chamber height of the preparation chambers 130a to 130d to be smaller than the chamber height of the medium chambers 132a to 132d. Due to this configuration, it is possible to prevent the sample from flowing into the medium chamber by capillary phenomenon in the preparation chamber itself during sample injection.
  • the trash chambers 133a to 133d are preferably hydrophilic coated, and the hydraulic burst valves 71a to 71d installed on the outside are the "magnetic burst valve", “hydrophobic burst valve” and “laser burst” described above. It can be noted that any one or more of “valve” may be replaced.
  • the chamber height of the thresh chambers 133a to 133d it is preferable to design the chamber height of the thresh chambers 133a to 133d to be smaller than the chamber height of the discharge chambers 132a to 132d. Due to this configuration, the debris and excess sample once moved to the tresh chamber can be prevented from flowing back to the medium chamber by capillary action in the trech chamber itself. 13 is a detailed view of the hydraulic burst valve 71b.
  • the inoculation of the medium 80 by applying the surface of the medium 80 with a sample and moving the excess sample to the tresh chamber is referred to as "coating inoculation".
  • coating inoculation By the application inoculation, it is possible to automate the streaking operation that has been done by conventional manual work.
  • the sampling rod 300a is separated from the sampler 300 (step 1).
  • Reference numerals 63 and 63a provide a screw fastening portion to be detachable between the sampling rod 300a and the culture tube 300b.
  • a cotton swab (60) for collecting the specimen from the inspection site. The user collects a sample using a swab 60 from a suspected place or inspection site 55.
  • the sampling rod (300a) and the culture tube (300b) is coupled through a screw fastening portion (63, 63a) and incubated for 24 hours enrichment (step 2).
  • the culture tube (300b) is enriched culture medium 69 is stored to help the growth of microorganisms.
  • the enrichment broth is preferably peptone water.
  • the culture cap 67 is separated from the sampler 300 (step 3).
  • Reference numerals 67 and 67a provide a screw fastening portion to be detachable between the culture cap 67 and the culture tube 300b.
  • the enriched sample in the culture tube 300b is transferred through the sample inlet 121 of the disc 100 (step 4).
  • the enriched sample in the culture tube 300b is transferred to the preparation chambers 130a to 130d.
  • Reference numeral 69 serves to filter the foreign matter in the enrichment culture medium 69 to prevent the transfer to the disk 100 by a filter.
  • the specimen stored in the preparation chambers 130a to 130d is moved into the medium chambers 132a to 132d by opening the burst valves 70a to 70d.
  • the surface of the medium in the medium chamber is coated with a sample by the coating inoculation operation.
  • burst valves 71a to 71d are opened to transfer the surplus specimens in the medium chamber to the trech chambers 133a to 133d.
  • microorganisms are cultured in a media chamber for 24 hours.
  • the medium chamber is read by the image sensor, and the colony is counted by the central controller to qualitatively and quantitatively analyze the microorganism.
  • test result according to the reading result is optionally displayed on the computer monitor, and remotely connected and remotely transmitted via the internet network automatically or manually.
  • the remote site calculates the user's hygiene score based on the inspection result and retransmits the user's hygiene score to the input / output device.
  • the hygiene score is preferably calculated based on the number of tests and the colony density of microorganisms.
  • 15 is an example showing the result of culturing microorganisms in the medium chamber.
  • Each spot represents a colony of microorganisms and different colors for each microorganism when chromogenic media is used.
  • the microbial analysis apparatus 100 determines whether the user ejects or stops the disk. If the user ejects or stops the disk, the microbial analysis apparatus continues the analysis while ignoring it. At this time, a warning message is notified to the user or a password is required. If the password is correct, the user will be asked to eject or stop the disk.
  • the memory of the wireless RF IC 188 stores a disk usage history, expiration date information or information on the types of microorganisms that can be analyzed.
  • the barcode pattern stores the identification information or the expiration date information of the disk or the type of microorganism to be analyzed.
  • the expiration date information informs the user that the expiration date is also unavailable for the disc.
  • FIG. 16 is a toploading microorganism analysis incorporating a thermostat or a temperature control device 730 for maintaining an optimal constant temperature during enrichment of microorganisms in the sampler 300 in step 2 of FIG.
  • a thermostat or a temperature control device 730 for maintaining an optimal constant temperature during enrichment of microorganisms in the sampler 300 in step 2 of FIG.
  • One embodiment of the apparatus 200 is shown.
  • the enrichment culture is preferably made by inserting the culture tube 300b of the sampler 300 into a thermostat or a temperature control device 730.
  • the microorganism analysis apparatus 200 of the disk may be loaded by opening the cover 751 for top loading and fitting the disk 100 to the turntable 181.
  • FIG. 17 is a view showing an embodiment of a front loading type microorganism analyzing apparatus incorporating a thermostat or a temperature control device, and the temperature sensor 731 to maintain an optimal constant temperature during enrichment of microorganisms.
  • the enrichment culture is preferably made by front loading by inserting the void 171 of the body 100 to the turntable 181 in a thermostat or temperature control device 730.
  • Reference numeral 733 is a transparent window, which allows to observe the culture state from the outside during the culture period.
  • the microbial analysis apparatus 200 may include an analysis start button 745 and a stop button 746.
  • reference numeral 742 denotes a power on / off button of the microbial analysis apparatus
  • reference numeral 741 denotes an LED for indicating a power state.
  • Reference numeral 760 denotes a display device for displaying a progress state of the microorganism analyzing apparatus 200, and a liquid crystal display (LCD) is preferred as the display device.
  • LCD liquid crystal display
  • the display device 760 displays an analysis result or displays a progress state according to a main process of the microbial analysis device.
  • the display device 760 may display the progress according to the main process (prep process, enrichment process, culture process) and steps in the form of a percentage (%) or a bar graph.
  • the display device 760 may provide a graphic user interface.
  • the graphical user interface preferably allows the user to provide temperature settings, incubation time settings for the thermostat. It is also preferred to notify the user via an alarm when the incubation time has elapsed.
  • the temperature control device applies a carbon or carbon nanotube, which is a high resistance material, onto a heater 761 in the tray 761 by applying a carbon or carbon nanotube, which is a high resistance material, on a heater or a PET (Polyethylene Terephthalate) film. It is preferred to include.
  • the microbial analysis device is further provided with calculation software for quantifying negative, positive or microbial levels for a specific microorganism by reading and analyzing an image of the medium chamber.
  • the cover means is installed at the outermost and innermost circumferences of the body 100. It is preferred that the cover means is provided with a permanent magnet 820b in the lower body 2 and a ferromagnetic material 820a in the upper body 1 to be closed by the force between the upper body and the lower body.
  • the solid medium in the medium chambers 132a to 132d contains a large amount of water. Unlike in FIG. 16, if the culture is carried out straight with the medium installed in the lower body 2, evaporation occurs, resulting in moisture and water droplets on the ceiling of the upper body 1, and the drop of water falls on the medium, thereby destroying the culture state. Not only is this possible, the medium can quickly dry out by evaporation during cultivation.
  • the medium 80 is installed on the upper body 1 side, as shown in FIG.
  • the sample inlet (121a to 121d) is installed on the upper body (1) side.
  • the medium in the medium chamber is preferably a solid medium to an agar medium.
  • Reference numeral 335 denotes a reference hole indicating a reference of the body.
  • Another aspect of the invention is characterized in that the thin film adhesive tape 2a and the ferromagnetic material 820a are replaced by a magnetic sheet to constitute the cover means.
  • one side of the magnetic sheet is coated with a pressure-sensitive adhesive is preferably attached to the upper body (1).
  • the permanent magnet 820b in the lower body 2 acts on the magnetic sheet on the upper body 1 so that the upper body 1 and the lower body 2 are coupled by a magnetic force to form a single body. (100).
  • the magnetic sheet serves as a thin film adhesive tape and a ferromagnetic material at the same time. That is, the magnetic sheet has an advantage of providing the assembly means and the cover means of the upper body 1 and the lower body 2 at the same time.
  • the magnetic sheet is preferably a combination of a cushion rubber material and magnetic powder in the form of a thin film.
  • the cushion rubber material increases the adhesion when the upper body 1 and the lower body 2 are coupled by magnetic force.
  • the magnetic sheet is black, the absorbance of the laser light is high, which is very easy to construct the laser burst valve.
  • the magnetic sheet is preferably in the form of a thin film having a thickness of 0.05 mm to 0.1 mm.
  • FIG 19 shows one embodiment of the opening means using an electromagnet board.
  • the body 100 is attached to and detached from the seating portion 332b of the electromagnet board 330. It is preferred to follow the shape of the body 100 with a seating edge 332a.
  • the body 100 Before the lower body 2 is separated from the body 100, the body 100 is seated on the seating portion 332b of the electromagnet board 330.
  • the body 100 unlike the arrangement at the time of cultivation (see Fig. 16) as shown in Figure 17, it is preferred to seat by changing the top and bottom arrangement of the lower body and the upper body.
  • Reference numeral 182 denotes a fixed table for placing the body 100, and is top loaded onto the fixed table through the center void 170. It is shown that the body has a groove 182b to be mechanically engaged with the fixed table 182.
  • the community gathering process for identification experiments proceeds as follows. First, the body 100 is fitted into the groove 182b, and then the power button 331 is turned on, and then separated from the body 100 into the lower body 2 to collect a sample of the suspected colony from the medium 80. . This collected community will be used for identification experiments. After the power button 331 is off (off), the lower body 2 is coupled to the upper body in accordance with the reference pillar 334. Thereafter, when the knobs 182b and the knobs are pressed, the central void 170 fastened to the groove 182b is released to remove the body 100 from the electromagnet board 330. In addition, the rim groove 333 on the seating edge 332a allows the body 100 to be easily pulled out of the electromagnet board 330.
  • the reference column 334 is fitted to fit the reference hole 335 to set the standard when the body is mounted on the electronic board and recombination of the upper body and the lower body.
  • FIG. 20 shows an embodiment of an oxygen supply valve for supplying oxygen to the aerobic microorganisms in the medium chamber.
  • the oxygen supply valve is composed of a hole (10b) for allowing the entry and exit of oxygen, the connecting passage 135 and the stopper (520a) connected to the medium chambers (132a to 132d).
  • the body 100 When using the microbial analysis device, the body 100 is rotated at a high speed, at which time the stopper 520a beats the adhesive strength of the adhesive means 521a by the strong centrifugal force acting on the stopper 520a and leaves the pores ( 10b) is opened.
  • the hole 10b is closed by the adhesive force (or the pressing force) between the stopper 520a and the bonding means 521a, and the stopper 520a is attached by the centrifugal force. It is characterized in that it is opened while leaving the adhesive strength (adhesive strength) with the means 521a.
  • the adhesive means 521a is preferably a double-coated tape or a rubber coating material having a cushion.
  • the plug is preferably a solid thin film cylinder. In this case, only the upper surface of the stopper was brought into close contact with the hole 10b with a double-sided tape 521a.
  • the portion of the pores 10b is preferred to use as a cushioned rubber or silicone material to increase the closure of the pores 10b.
  • the stopper may be a permanent magnet of a thin film cylindrical shape and a ferromagnetic material may be used as the stopper.
  • the hole is closed by the attraction force of the stopper and the ferromagnetic material, and the stopper 520a overcomes the attraction force and escapes due to the strong centrifugal force acting on the stopper by the high-speed rotation of the body 100 during use. Is opened.
  • 21 is an embodiment of a microbial analysis apparatus using a test strip.
  • Reference numeral 145 denotes a test strip using a membrane.
  • test strip is preferably immobilized with an antibody on a Nitrocellurose membrane or a PVDF membrane, which is referred to as a test line hereinafter.
  • the antibody of the test line is preferably an antibody that specifically binds to at least one selected from Salmonella, Listeria, and O157.
  • the test strip also preferably includes a sample pad 41a, a conjugate pad 41b, and an absorbent pad 41c.
  • the conjugate pad is preferably one in which a gold conjugate, which is complexed with gold particles, is deposited in a lyophilized form.
  • the test strip is characterized in that the reaction test by immunochromatography.
  • the immunochromatography method is a combination of immunochemistry and chromatography (Chromatogrphic Assay), the specific immunoreactivity of the antibody to the antigen, the color development characteristics of the colloidal gold and porous membrane (Porous) It is a test method that applies the movement of molecules by capillary phenomenon of membrane).
  • the test strip may further include a reference line, and the reaction concentration of the reference line may be set to a cutoff value to facilitate the discrimination of negative or positive reactions.
  • Qualitative or quantitative analysis may be performed based on the difference or ratio of the reaction intensity between the reference line and the test line.
  • a sample injection port 121 for injecting a sample
  • a prep chamber 130 in which an antibody is immobilized to perform a specific binding reaction with the sample injected from the sample injection port; After sending the waste which did not bind with the antibody in the prep chamber to the trech chamber 133, and sending the microorganism that caused the reaction with the antibody to the enrichment chamber 131, papain digest (papain).
  • digestion digestion chamber 134 containing reagents for carrying out mecaptoethanol reduction or pepsin digestion;
  • An enrichment chamber 131 for storing a liquid culture medium for enriching microorganisms transferred from the preparation chamber 130 by the digestion;
  • a strip chamber 132 including a test strip 145 to which at least one antibody for immobilizing an antibody antigen reaction against the enriched microorganism is immobilized;
  • a trace chamber 133 for collecting debris from the strip chamber 132; And valves (64, 65, 66, 68) for controlling liquid movement between the chambers;
  • a flow path (channel) for providing a movement path of the specimen; And a rotatable body 100 in which the flow path, the valve and the chamber are integrated.
  • the antibody is cleaved by the reagent that performs pepsin digestion, is separated from the preparation chamber 130, and transferred to the enrichment chamber 131.
  • the body 100 may include one or more strip chambers 132, and may be modified to independently analyze different microorganisms.
  • the body 100 may include one or more strip chamber 132 and the preparation chamber 130, it may be modified to independently analyze different microorganisms.
  • the preparation chamber 130 is characterized in that it comprises magnetic beads (magnetric bead) or gold coated magnetic beads.
  • the antibody is immobilized on the surface of the magnetic beads.
  • the magnetic beads are preferably of superparamagnetic or ferromagnetic properties. It is also preferred that the diameter of the outlet channel 130f of the prep chamber 130 is smaller than the diameter of the magnetic bead so that the magnetic bead stays in the prep chamber. In the present invention, the diameter of the magnetic beads is preferably 0.1um to 10um.
  • the movable sensor 5b is characterized in that the image sensor 103 performs a "search process of the stream chamber" before the image of the strip chamber 132 is taken.
  • a permanent magnet 5c is preferably provided on the body 100 to facilitate optical alignment between the image sensor 103 and the strip chamber 132. .
  • Reference numeral 170 denotes a disk gap.
  • Reference numeral 335 denotes a reference hole indicating a reference of the body.
  • Reference numeral 188 denotes the above-described memory embedded wireless RF IC or electronic tag device.
  • a solid medium or agar medium may be used instead of the test strip in the strip chamber.
  • the microorganisms specifically bound to the antibody in the preparation chamber 130 is transferred to the enrichment chamber 131, and then cultured on the solid medium or agar medium, so that the reliability of separation culture is greatly improved.
  • FIG. 21 shows that only antibodies having a specific binding to the microorganisms are cultured in the enrichment chamber 131, thereby greatly increasing the selectivity for the microorganisms, and thus performing immunological identification tests collectively.
  • One embodiment is provided that provides advantages.
  • the microorganism analyzing apparatus of the present invention and an analysis method using the same are suitable for a thin film type apparatus for diagnosing and detecting a small amount of microorganisms present in a sample for contaminants such as food poisoning test, food microbial test, and water quality.

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Abstract

La présente invention concerne un système d'analyse de micro-organisme qui peut détecter un micro-organisme présent dans un aliment, le lait, les produits laitiers, les produits agricoles et marins, les boissons et les échantillons environnementaux par la réaction biochimique de sucre contenu dans un milieu et d'une enzyme contenue dans un micro-organisme en utilisant l'un quelconque ou plusieurs milieux parmi un milieu chromogène présentant une couleur spécifique selon l'état de culture d'une certaine bactérie, un milieu à base d'agar, un milieu différentiel et un milieu sélectif. L'invention concerne également un procédé d'analyse utilisant ce système. Plus spécifiquement, la présente invention concerne un système d'analyse de micro-organisme comprenant : une ouverture d'injection d'échantillon pour injecter un échantillon ; un échantillonneur qui récolte un échantillon à partir d'une zone devant être examinée et injecte l'échantillon récolté dans l'ouverture d'injection d'échantillon ; une chambre de préparation pour stocker temporairement l'échantillon injecté à partir de l'ouverture d'injection d'échantillon ; une chambre d'enrichissement qui comprend un milieu de culture liquide pour obtenir l'échantillon à partir de la chambre de préparation et pour enrichir un micro-organisme présent dans l'échantillon ; une ou plusieurs chambres de milieu de culture qui fournissent un milieu de culture au micro-organisme enrichi pour produire des nutriments ; une chambre de déchets pour récolter les restes et un excès d'échantillons ; une soupape pour déplacer le fluide et l'échantillon entre les chambres ; un canal fournissant un trajet de déplacement de l'échantillon ; et un corps en forme de disque pivotant dans lequel le canal, la soupape et les chambres sont intégrés et un procédé d'analyse l'utilisant.
PCT/KR2010/006266 2009-09-22 2010-09-14 Système d'analyse de micro-organisme, et procédé d'analyse l'utilisant WO2011037348A2 (fr)

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KR1020090090318A KR101209277B1 (ko) 2009-09-22 2009-09-22 미생물 분석 장치 및 이를 이용한 분석 방법

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107663504A (zh) * 2017-10-09 2018-02-06 宜昌喜旺食品有限公司 一种智能检测牛奶细菌数量的设备及检测方法
EP3660160A1 (fr) * 2018-11-28 2020-06-03 GTZ Microlab Detect, S.L. Dispositif portable et procédé de détection de micro-organismes ou de métabolites dans un échantillon
WO2023133714A1 (fr) * 2022-01-12 2023-07-20 广州工商学院 Dispositif de mesure de quantité fongique et procédé de mesure pour tester la qualité des yaourts

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CN113999813A (zh) * 2021-10-12 2022-02-01 云南和泽西南生物科技有限公司 一种脐带间充质干细胞原代培养方法

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* Cited by examiner, † Cited by third party
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DE3410990C2 (de) * 1984-03-17 1986-04-03 Biotest-Serum-Institut Gmbh, 6000 Frankfurt Vorrichtung zur Prüfung der Luft auf ihren Keimgehalt
FR2677664A1 (fr) 1991-06-13 1992-12-18 Millipore Sa Dispositif et procede de controle microbiologique des liquides sous pression.
WO1999047637A1 (fr) * 1998-03-19 1999-09-23 Amanzi Technologies Limited Analyse microbiologique d'un echantillon de liquide
DE19943853A1 (de) 1999-09-13 2001-03-15 Udo Hoelker Bioreaktor zur Fermentierung von festen Stoffen

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107663504A (zh) * 2017-10-09 2018-02-06 宜昌喜旺食品有限公司 一种智能检测牛奶细菌数量的设备及检测方法
CN107663504B (zh) * 2017-10-09 2024-03-19 宜昌喜旺食品有限公司 一种智能检测牛奶细菌数量的设备及检测方法
EP3660160A1 (fr) * 2018-11-28 2020-06-03 GTZ Microlab Detect, S.L. Dispositif portable et procédé de détection de micro-organismes ou de métabolites dans un échantillon
WO2020109457A1 (fr) * 2018-11-28 2020-06-04 Gtz Microlab Detect, S.L. Dispositif portable et procédé de détection de micro-organismes ou de métabolites dans un échantillon
WO2023133714A1 (fr) * 2022-01-12 2023-07-20 广州工商学院 Dispositif de mesure de quantité fongique et procédé de mesure pour tester la qualité des yaourts

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WO2011037348A3 (fr) 2011-07-14
KR101209277B1 (ko) 2012-12-12

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