WO2017191715A1 - Instrument de test de chromatographie d'acides nucléiques, kit de test de chromatographie d'acides nucléiques et procédé d'utilisation d'un instrument de test de chromatographie d'acides nucléiques - Google Patents

Instrument de test de chromatographie d'acides nucléiques, kit de test de chromatographie d'acides nucléiques et procédé d'utilisation d'un instrument de test de chromatographie d'acides nucléiques Download PDF

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WO2017191715A1
WO2017191715A1 PCT/JP2017/010554 JP2017010554W WO2017191715A1 WO 2017191715 A1 WO2017191715 A1 WO 2017191715A1 JP 2017010554 W JP2017010554 W JP 2017010554W WO 2017191715 A1 WO2017191715 A1 WO 2017191715A1
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nucleic acid
strip
acid chromatography
chromatography test
test instrument
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PCT/JP2017/010554
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English (en)
Japanese (ja)
Inventor
秀彦 西塚
充裕 吉田
恵美子 小菅
深澤　眞
伊藤　好久
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東洋製罐グループホールディングス株式会社
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Publication of WO2017191715A1 publication Critical patent/WO2017191715A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
    • 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
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • 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
    • G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements

Definitions

  • the present invention relates to a nucleic acid chromatography technique, and in particular, a nucleic acid chromatography test instrument, a nucleic acid chromatography test kit, and a nucleic acid chromatography test instrument that detect a target nucleic acid using a strip for genetic testing on which a probe is immobilized. Regarding usage.
  • a target nucleic acid is hybridized (complementarily bound) to a probe immobilized on a strip using a strip for genetic testing in which the probe is immobilized.
  • the detection was performed. Specifically, for example, genomic DNA is extracted from a sample to be tested, and a genetic test strip is immersed in a reaction solution containing an amplification product obtained by amplifying a target nucleic acid by a PCR (polymerase chain reaction) method or the like. Then, the target nucleic acid is detected by developing the amplification product on a strip and hybridizing with the probe, and visually confirming the labeling dye that binds to the amplification product hybridized with the probe.
  • the extracted genomic DNA was placed in a PCR tube together with a PCR reaction reagent and PCR was performed, and then a chromatographic developing solution containing a labeling dye was mixed in the PCR tube to color the amplification product. Then, as shown in FIG. 5, the gene test strip is immersed in the reaction solution in the PCR tube, the reaction solution is spread on the strip by capillary action, and the amplification product hybridized to the probe immobilized on the strip is used. It was done to check the colored lines shown.
  • an analyte detection device described in Patent Document 1 can be exemplified.
  • the amplification product can be spread on a strip in a device configured to be a semi-closed system. Specifically, by inserting a container containing a liquid sample into the cavity of the device, the lower part of the container is pierced so that the sample in the container is released to reach the strip and deployed.
  • a device can be suitably used when the conventional nucleic acid chromatography protocol described above is performed.
  • the chromatography development solution containing the labeling dye is later developed on the strip, and the amplification product hybridized with the probe.
  • a coloration protocol for. According to such a protocol, the probe and the amplification product can be hybridized first, and then the labeling dye can be bound to the amplification product, so that the accuracy of hybridization can be further improved and stable coloring can be achieved. As a result, the detection accuracy can be improved.
  • the above-described conventional device is not suitable for the case where the PCR reaction solution and the chromatographic developing solution are separately developed, and it has been difficult to use in such a protocol.
  • the present inventors have conducted intensive research and can perform nucleic acid chromatography in a semi-sealed system, and after developing the PCR reaction solution on the strip, the chromatography developing solution is separately developed on the strip.
  • the present invention has been completed by developing a nucleic acid chromatography test instrument that can be used. That is, the present invention can suppress contamination, reduce the amount of a PCR reaction solution containing a target nucleic acid used for detection, and improve the detection accuracy, a nucleic acid chromatography test instrument, a nucleic acid chromatography test kit, And it aims at providing the usage method of a nucleic acid chromatography test
  • a nucleic acid chromatography test instrument of the present invention is a nucleic acid chromatography test instrument that holds a strip for genetic testing and detects a target nucleic acid using the strip, and inserts a PCR tube.
  • An injecting portion having a first inlet possible and a second inlet for injecting a chromatographic developing solution, a developing portion covering the detection region of the strip, and an incision supporting an incision means for incising the tube
  • a means supporting portion and a strip holding portion for holding the strip are provided.
  • the nucleic acid chromatography test kit of the present invention comprises the above-described nucleic acid chromatography test instrument, at least a PCR reaction reagent containing a primer mix comprising one or more primer sets, at least a labeling dye and a buffer solution. And a chromatographic developing solution.
  • the method for using the nucleic acid chromatography test instrument of the present invention includes the step of inserting the tube in which the reaction liquid containing the target nucleic acid is sealed into the first inlet of the nucleic acid chromatography test instrument.
  • a probe having a complementary base sequence immobilized on the strip by incising the tube by means, allowing the reaction solution to permeate the strip from the tube, and developing the strip.
  • the present invention in performing nucleic acid chromatography, contamination can be suppressed, the amount of PCR reaction solution containing the target nucleic acid used for detection can be reduced, and detection accuracy can be further improved.
  • FIG. 1 It is a perspective view which shows the external appearance of the nucleic acid chromatography test
  • nucleic acid chromatography test instrument a nucleic acid chromatography test kit
  • nucleic acid chromatography test instrument of the present embodiment the nucleic acid chromatography protocol applied in the method for using the nucleic acid chromatography test instrument, the nucleic acid chromatography test kit, and the nucleic acid chromatography test instrument of the present embodiment will be described.
  • the target nucleic acid moves (develops) in the genetic test strip in a certain direction, so that it is captured by the probe spotted on the strip, and at the spot where the probe is immobilized, By being concentrated, color development up to a level that can be visually observed by the labeling dye bound to the target nucleic acid is obtained, and the target nucleic acid is detected.
  • the amplification process of the target nucleic acid is performed by the PCR method or the like. That is, the amplification product by the PCR method or the like consists of a target nucleic acid. Specifically, a PCR reaction reagent containing a primer set for amplifying the target nucleic acid in the genomic DNA of the organism to be examined is injected into the PCR tube. As the primer set, a primer mix comprising one or more primer sets can be used, and multiplex PCR for simultaneously amplifying a plurality of target nucleic acids can also be performed.
  • the PCR reaction reagent includes a primer set, a nucleic acid synthesis substrate (for example, dNTPmixture (dCTP, dATP, dTTP, dGTP)), a nucleic acid synthase (for example, EX Taq HotStart DNA polymerase), a buffer, and the like.
  • a nucleic acid synthesis substrate for example, dNTPmixture (dCTP, dATP, dTTP, dGTP)
  • a nucleic acid synthase for example, EX Taq HotStart DNA polymerase
  • the PCR tube is injected with such a PCR reaction reagent, genomic DNA extracted from a biological sample, and a solution containing water as the remaining component, and this solution is used as a reaction solution used for PCR.
  • Genomic DNA can be extracted by a general method such as a method using a CTAB method (Cetyl trimethyl ammonium bromide) or a method using a DNA extraction apparatus.
  • a general thermal cycler etc. can be used as a PCR apparatus.
  • the PCR reaction can be performed, for example, under the following reaction conditions, but can be performed with appropriate changes, and the whole may be performed in about 15 to 30 minutes.
  • a PCR reaction solution containing an amplification product (target nucleic acid) obtained by PCR is developed on a genetic test strip.
  • the genetic test strip is an elongated member on which a probe for capturing an amplification product is immobilized, and is formed so that the solution penetrates the entire member by capillary action and is easily developed.
  • cellulose such as cellulose membrane is preferably used as the strip member.
  • a material having a resin adhered to one surface thereof can be suitably used.
  • a genetic test strip In a genetic test strip, one or two or more probes are penetrated (impregnated) with a detection region fixed in a line in the short direction of the strip and a PCR reaction solution or a chromatographic development solution at a specific position of the strip. And a development start area which is a part where development starts. It should be noted that the deployment start area may be a very small part of the strip.
  • one or more regions (spots) to which the probe is immobilized are formed (spotted) in a line shape in the short direction of the strip.
  • the number of lines on which the probes are immobilized is not particularly limited, and for example, a line on which 8 to 12 lines or the like are formed can be suitably used.
  • a blue labeling dye is bound to the amplification product hybridized to the probe immobilized in such a line shape, the blue line can be visually detected on the detection region of the strip.
  • other color lines for example, red lines, are preferably arranged as positioning markers in the detection region.
  • the labeling dye is not particularly limited.
  • a labeling dye using an avidin / biotin system can be preferably used.
  • the development start region is a portion where the PCR reaction solution and the chromatographic development solution are first soaked.
  • the development start area is continuously formed adjacent to the detection area, and the liquid soaked in the development start area develops to the detection area by capillary action. It is also preferable to continuously form the water absorption promotion region adjacent to the detection region on the side opposite to the development start region so that the liquid can easily penetrate into the strip.
  • the water absorption promotion region can be configured by, for example, laminating a member such as a water absorption pad or cellulose on the strip, and the water absorption of the water spread on the strip can be promoted by such a water absorption member.
  • the strip can be made easier to handle by forming a region in which the thickness is increased by laminating members on the strip.
  • the PCR reaction solution is infiltrated into the development start region in such a genetic test strip.
  • the PCR reaction solution is developed from the development start region to the detection region.
  • an amplification product having a base sequence complementary to the probe immobilized on the detection region is contained in the PCR reaction solution, the probe and the amplification product are hybridized.
  • multiple types of probes are individually immobilized on a strip in a line shape, amplification products that hybridize to each probe are obtained by multiplex PCR and included in the PCR reaction solution. The corresponding probe and the amplified product are hybridized.
  • a chromatographic developing solution is impregnated into the development start region of the genetic test strip.
  • the chromatographic developing solution is desirably soaked after the PCR reaction solution has sufficiently developed to the detection region or the water absorption promoting region. This makes it possible to sufficiently soak the chromatography developing solution into the detection region.
  • the chromatography developing solution is preferably soaked into the development start region after, for example, about 5 to 10 minutes have elapsed after the PCR reaction solution has been developed into the detection region.
  • the chromatographic developing solution contains at least a labeling dye (colored beads such as those produced by an avidin / biotin system) and a buffer solution. This labeling dye can bind to the amplification product.
  • the developing solution is developed from the development start region to the detection region.
  • the labeling dye contained in the developing solution binds to the amplified product hybridized with the probe, whereby a colored line appears at the position where the probe is immobilized.
  • the target nucleic acid can be detected by confirming the colored line on the strip.
  • a chromatographic development solution containing a labeling dye is later developed on the strip and hybridized with the probe.
  • the amplified product can be colored. For this reason, a colored line can be obtained stably and detection accuracy can be improved.
  • the probe and the amplification product are hybridized first, and then the labeling dye is bound to the amplification product. Therefore, the labeling dye is bound to the amplification product prior to the hybridization. This is considered to be because the steric hindrance of hybridization assumed in the case of the reaction is suppressed, and the accuracy of hybridization can be further improved.
  • FIG. 1 shows a perspective view of the nucleic acid chromatography test instrument of the present embodiment.
  • the nucleic acid chromatography test instrument of the present embodiment includes a first inlet 111 into which the PCR tube 5 can be inserted, and a second inlet 112 for injecting a chromatography developing solution.
  • the injection part 11 provided, the development part 12 covering the detection region in the genetic test strip 4, the incision means support part 21 for supporting the incision means 3 for incising the PCR tube 5, and the strip for holding the genetic test strip 4
  • a holding part 22 is provided.
  • the nucleic acid chromatography test instrument of this embodiment has a main body 1 having an injection part 11 and a developing part 12, a cutting means support part 21 and a strip holding part 22, and is fitted to the main body part 1.
  • a configuration including the bottom lid 2 is also preferable.
  • the nucleic acid chromatography test instrument of the present embodiment is formed by integral molding instead of separately configuring the main body 1 and the bottom lid 2.
  • the slit can be provided, for example, on any of the four side walls of the inspection instrument. Then, the genetic test strip 4 is inserted into the test instrument through the slit and disposed in the strip holding unit 22.
  • FIG. 1 shows such a nucleic acid chromatography test instrument.
  • the first inlet 111 into which the PCR tube 5 can be inserted and the second inlet 112 for injecting the chromatographic developing solution are separately provided. It is configured. For this reason, it is possible to suitably perform this application protocol in which the above-described PCR reaction solution is first developed on the strip, and then the chromatography development solution containing the labeling dye is later developed on the strip.
  • first injection port 111 and the second injection port 112 are formed in the longitudinal direction of the strip above the development start region in the genetic test strip 4.
  • the PCR tube 5 is inserted from the first inlet 111 and the PCR tube 5 is incised by the incision means 3, so that the reaction solution can be infiltrated into the strip from the PCR tube 5 and developed.
  • the labeling dye is infiltrated into the strip and developed, and the amplified product that has been developed and hybridized to the probe A labeling dye can be attached. For this reason, it is possible to suitably perform the protocol applied to the present embodiment.
  • the arrangement order of the first inlet 111 and the second inlet 112 is not limited to that shown in FIG. Further, by forming the first inlet 111 and the second inlet 112 side by side in the longitudinal direction of the genetic test strip 4 in this way, a linked nucleic acid chromatography test corresponding to a linked PCR tube described later is performed. It is also possible to make it suitable for constructing an instrument.
  • the detection region of the genetic test strip 4 can be easily confirmed visually.
  • at least one of the development part 12 covering the detection region in the genetic test strip 4 or the strip holding part 22 for holding the genetic test strip 4 is made of a transparent member.
  • all of the injection part 11, the development part 12, the incision means support part 21, and the strip holding part 22 in the nucleic acid chromatography test instrument of the present embodiment can be made of a transparent member.
  • an acrylic resin such as polystyrene, polycarbonate, polyethylene terephthalate (PET), methyl methacrylate (MMA), or the like can be suitably used.
  • the nucleic acid chromatography method can be performed with the above-described application protocol using the general genetic test strip 4 and the PCR tube 5 with higher accuracy. It becomes possible to carry out.
  • FIG. 2 is a plan view and a side view of the nucleic acid chromatography test instrument according to the present embodiment
  • FIG. 3 is a plan view and a side view of the bottom lid of the test instrument
  • FIG. It is a side view of an incision means.
  • the nucleic acid chromatography test instrument of the present embodiment includes a main body portion 1 having an injection portion 11 and a development portion 12 having a first injection port 111 and a second injection port 112, and a PCR tube.
  • 5 has an incision means support portion 21 for supporting the incision means 3 for incising 5 and a strip holding portion 22 for holding the genetic test strip 4, and a bottom lid 2 fitted to the main body portion 1.
  • the incision means support portion 21 is provided so that at least a part of the incision means 3 is disposed in the first injection port 111.
  • the PCR tube 5 is filled with an amplification product (target nucleic acid) obtained by PCR. After performing PCR, the PCR tube 5 is pushed into the first inlet 111 without opening the lid. At this time, the PCR tube 5 is cut, perforated, incised, or the like by the incision means 3, and a crack or an opening is generated at the bottom or side of the PCR tube 5. Then, the reaction solution in the PCR tube 5 flows into the genetic test strip 4. Thereby, the PCR reaction solution soaks into the genetic test strip 4 and develops.
  • an amplification product target nucleic acid
  • the nucleic acid chromatography test instrument of the present embodiment can develop the reaction solution into a strip by opening a crack in the bottom or side surface of the PCR tube 5 without opening the lid. At this time, the inspection instrument inserts the PCR tube 5 into the first inlet 111, thereby closing the opening of the first inlet 111 and opening only the second inlet 112. It has become. For this reason, according to the nucleic acid chromatography test instrument of this embodiment, it is possible to reduce the risk of contamination.
  • the size of the opening of the second inlet 112 is preferably smaller than the size of the opening of the first inlet 111. As a result, the risk of contamination can be further reduced.
  • the shape of the first inlet 111 is not particularly limited, and can be formed, for example, in a substantially thin conical column or elliptical truncated cone, and the horizontal plane opening of the first inlet 111 is, for example, a perfect circle. Can be formed.
  • the shape of the second injection port 112 is not particularly limited, and can be formed, for example, in a substantially thin conical column having a narrow shape.
  • the horizontal plane opening of the second injection port 112 is, for example, a perfect circle. It can be formed as follows.
  • the first inlet 111 can be formed so that the central axis of the first inlet 111 is inclined with respect to the vertical direction.
  • the central axis is inclined in the longitudinal direction of the genetic test strip 4 to the opposite side with respect to the detection region to form the first injection port 111, and the PCR tube 5 is obliquely inserted into the inclined first injection port 111.
  • the incision position of the PCR tube 5 by the incision means 3 may be adjusted as appropriate.
  • the second inlet 112 can be formed so that the central axis of the second inlet 112 is inclined with respect to the vertical direction.
  • the horizontal plane opening of the first inlet 111 so as to have a track shape (a shape similar to a track of an athletics), for example, two opposing linear sides are opposed to each other. It is preferable to use a semicircular shape (first shape).
  • the track shape may include a shape (second shape) formed by two semi-circles that are convex on the inner side facing two opposite sides.
  • the PCR tube 5 becomes linear as the PCR tube 5 is pushed into the first inlet 111. Cracks that are compressed and incised by the two sides of the surface (in the case of the first shape) or by two semicircular surfaces convex inward (in the case of the second shape) As a result, the reaction solution can be easily opened out from the PCR tube 5.
  • first inlet 111 and the second inlet 112 have different shapes, sizes, heights, and the like.
  • the second inlet 112 is sized so that the PCR tube 5 cannot be inserted, so that the PCR tube 5 can be inserted incorrectly. It becomes possible to prevent.
  • an aluminum seal can be attached to the first inlet 111 and / or the second inlet 112 and peeled off during use.
  • the strip holder 22 is preferably formed with a positioning column 221 composed of a plurality of protrusions for fixing the genetic test strip 4.
  • the number thereof is not particularly limited, but it is preferable to provide a plurality of oppositely on the bottom lid 2 so that the genetic test strip 4 does not move in the nucleic acid chromatography test instrument of the present embodiment.
  • six sets of twelve positioning columns 221 are formed.
  • the incision means 3 is not particularly limited as long as it can incise the PCR tube 5 in the first injection port 111.
  • a metal blade, a metal needle, a plastic blade, a plastic needle or the like can be used.
  • an incision blade 31 for incising the PCR tube 5 inserted into the first inlet 111, and a gene It is preferable to have a strip contact portion 32 that contacts the test strip 4.
  • the strip contact portion 32 is provided on the metal blade or the like in this way, the PCR reaction liquid flowing out from the cut PCR tube 5 can move to the genetic test strip 4 through the strip contact portion 32 and easily penetrates into the strip. Therefore, the development on the strip can be performed more reliably.
  • the injection part 11 is formed in a substantially rectangular parallelepiped shape, and the development part 12 is formed thin and long enough to hold the genetic test strip 4 inside.
  • the volumes of the inspection instrument can be reduced.
  • the shapes of the injection part 11 and the development part 12 are not limited to these, and the injection part 11 can be formed in, for example, a substantially elliptical column shape, or the development part 12 can be formed thicker. .
  • the nucleic acid chromatography test instrument of this embodiment is arranged in a state where a plurality of the above-described nucleic acid chromatography test instruments are coupled in the short direction of the genetic test strip 4, and the same number as the nucleic acid chromatography test instrument is connected.
  • Each tube of the connection type tube is connected to the first inlet 111 in the nucleic acid chromatography test instrument so as to be simultaneously insertable.
  • the nucleic acid chromatography test instrument of this embodiment is preferably formed by connecting, for example, eight nucleic acid chromatography test instruments described above in the short direction.
  • eight PCRs are simultaneously performed using a commonly used 8-series PCR tube, and target nucleic acid detection is simultaneously performed for each PCR reaction solution. Can be done.
  • the linked nucleic acid chromatography test instrument it is possible to reduce the time and effort of processing when performing a plurality of tests, and it is possible to reduce the risk of sample mix-up. Moreover, according to such a plurality of processes, it is possible to contribute to cost reduction. Needless to say, the number of nucleic acid chromatography test instruments to be connected is not limited to eight, and may be other numbers such as four or twelve.
  • the nucleic acid chromatography test instrument of the present embodiment it is possible to reduce the risk of contamination due to scattering of the PCR reaction solution as compared with the conventional open-type nucleic acid chromatography method. Instability of detection accuracy due to liquid volatilization or the like can be eliminated. Further, according to the nucleic acid chromatography test instrument of the present embodiment, the PCR reaction solution is developed on the strip and then the chromatography development solution is developed on the strip, as compared with the conventional semi-sealed device described above. It can be made suitable for this application protocol, and detection accuracy can be further improved. Furthermore, by using a linked nucleic acid chromatography test instrument, it is possible to reduce the risk of processing troubles, sample mistaking, and the like, and further contribute to cost reduction.
  • the nucleic acid chromatography test kit of the present embodiment includes the above-described nucleic acid chromatography test instrument, a PCR reaction reagent including a primer mix consisting of at least one or two or more primer sets, at least a labeling dye and a buffer. It is preferable to include a chromatography developing solution containing a liquid.
  • nucleic acid chromatography test kit according to the present embodiment is configured as described above, PCR can be easily executed, the PCR reaction solution can be developed on the strip, and then the chromatography development solution can be developed on the strip. For this reason, the amplification product hybridized with the probe can be colored, and the detection accuracy can be improved.
  • the PCR tube 5 in which the reaction solution containing the target nucleic acid is sealed is inserted into the first inlet 111 of the nucleic acid chromatography test instrument described above.
  • the PCR tube 5 is incised by the incision means 3, the reaction solution is permeated into the genetic test strip 4 from the PCR tube 5, and the target nucleic acid in the reaction solution is immobilized on the genetic test strip 4.
  • Hybridize with a probe having a complementary base sequence and after a lapse of a predetermined time, inject a chromatographic developing solution from the second inlet 112, and hybridize the labeling dye contained in the developing solution to the probe. It is preferable to use a method of binding to the target nucleic acid.
  • the PCR reaction solution can be easily developed from the PCR tube to the strip first using the semi-enclosed nucleic acid chromatography test instrument. Can do. Then, this can be developed into a strip by injecting a chromatographic developing solution. For this reason, after first hybridizing the amplification product to the probe on the strip, a labeling dye is bound to this amplification product, and the target nucleic acid is detected by visually recognizing the colored line indicated by the labeling dye. can do. Therefore, the accuracy of hybridization can be further improved, stable coloring is performed, and the detection accuracy can be improved.
  • a plurality of genetic test strips can be held in a single nucleic acid chromatography test instrument, and the test instrument can be appropriately changed to have a first inlet and a second inlet corresponding to each. Is possible.
  • the present invention can be suitably used for performing nucleic acid chromatography in order to suppress contamination and improve detection accuracy.
  • the contents of the documents described in this specification and the specification of the Japanese application that is the basis of Paris priority of the present application are all incorporated herein.

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Abstract

Selon la présente invention, il est possible, lors de la réalisation d'une chromatographie d'acides nucléiques, de supprimer la contamination et de réduire la quantité de liquide de réaction de PCR contenant les acides nucléiques cibles utilisé pour la détection de manière à améliorer la précision de détection. La chromatographie d'acides nucléiques est effectuée à l'aide d'un instrument de test de chromatographie d'acides nucléiques qui contient une bandelette 4 de test génique et qui utilise la bandelette pour détecter des acides nucléiques cibles, l'instrument comprenant : une partie d'injecteur 11 qui est pourvue d'une première embouchure d'injection 111, dans laquelle un tube de PCR 5 peut être introduit, et d'une deuxième embouchure d'injection 112, dans laquelle une solution de révélateur chromatographique est injectée ; une partie de révélateur 12 qui recouvre une zone de détection de la bandelette ; une partie 21 contenant un moyen de découpe qui supporte un moyen de découpe 3 pour ouvrir le tube tout en le coupant ; et une partie 22 de porte-bandelette pour contenir la bandelette.
PCT/JP2017/010554 2016-05-02 2017-03-16 Instrument de test de chromatographie d'acides nucléiques, kit de test de chromatographie d'acides nucléiques et procédé d'utilisation d'un instrument de test de chromatographie d'acides nucléiques WO2017191715A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2016-092534 2016-05-02
JP2016092534A JP2017200446A (ja) 2016-05-02 2016-05-02 核酸クロマトグラフィー検査器具、核酸クロマトグラフィー検査キット、及び核酸クロマトグラフィー検査器具の使用方法

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WO2017191715A1 true WO2017191715A1 (fr) 2017-11-09

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Cited By (4)

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Publication number Priority date Publication date Assignee Title
WO2018123464A1 (fr) * 2016-12-26 2018-07-05 東洋製罐グループホールディングス株式会社 Instrument de test de chromatographie d'acide nucléique, kit pour test de chromatographie d'acide nucléique et procédé d'utilisation d'un instrument de test de chromatographie d'acide nucléique
CN109557299A (zh) * 2018-11-12 2019-04-02 必欧瀚生物技术（合肥）有限公司 一种多通道仪器用检测装置及检测方法
WO2020173446A1 (fr) * 2019-02-25 2020-09-03 上海快灵生物科技有限公司 Tube à bandelette réactive de réaction biochimique et son procédé d'utilisation, et kit
WO2024162460A1 (fr) * 2023-02-02 2024-08-08 株式会社Tba Dispositif d'inspection d'acides nucléiques et son utilisation

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US20090181388A1 (en) * 2007-08-17 2009-07-16 Ustar Biotechnologies (Hangzhou) Ltd. Enclosed unit for rapid detection of a target nucleic acid amplification product
WO2012070618A1 (fr) * 2010-11-24 2012-05-31 株式会社カネカ Procédé de détection d'acide nucléique amplifié et dispositif de détection
JP2015512250A (ja) * 2012-03-21 2015-04-27 ヴィーセル， ソシエダッド リミターダ 親和性バイオアッセイにおいて分析物を検出するデバイス

Patent Citations (3)

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US20090181388A1 (en) * 2007-08-17 2009-07-16 Ustar Biotechnologies (Hangzhou) Ltd. Enclosed unit for rapid detection of a target nucleic acid amplification product
WO2012070618A1 (fr) * 2010-11-24 2012-05-31 株式会社カネカ Procédé de détection d'acide nucléique amplifié et dispositif de détection
JP2015512250A (ja) * 2012-03-21 2015-04-27 ヴィーセル， ソシエダッド リミターダ 親和性バイオアッセイにおいて分析物を検出するデバイス

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018123464A1 (fr) * 2016-12-26 2018-07-05 東洋製罐グループホールディングス株式会社 Instrument de test de chromatographie d'acide nucléique, kit pour test de chromatographie d'acide nucléique et procédé d'utilisation d'un instrument de test de chromatographie d'acide nucléique
CN109557299A (zh) * 2018-11-12 2019-04-02 必欧瀚生物技术（合肥）有限公司 一种多通道仪器用检测装置及检测方法
WO2020173446A1 (fr) * 2019-02-25 2020-09-03 上海快灵生物科技有限公司 Tube à bandelette réactive de réaction biochimique et son procédé d'utilisation, et kit
WO2024162460A1 (fr) * 2023-02-02 2024-08-08 株式会社Tba Dispositif d'inspection d'acides nucléiques et son utilisation

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