WO2021152780A1 - Ｄｎａ抽出方法及び爪加工装置 - Google Patents

Ｄｎａ抽出方法及び爪加工装置 Download PDF

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Publication number
WO2021152780A1
WO2021152780A1 PCT/JP2020/003466 JP2020003466W WO2021152780A1 WO 2021152780 A1 WO2021152780 A1 WO 2021152780A1 JP 2020003466 W JP2020003466 W JP 2020003466W WO 2021152780 A1 WO2021152780 A1 WO 2021152780A1
Authority
WO
WIPO (PCT)
Prior art keywords
nail
dna extraction
dna
extraction method
processed
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/JP2020/003466
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English (en)
French (fr)
Japanese (ja)
Inventor
麻生川　稔
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
NEC Corp
Original Assignee
NEC Corp
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by NEC Corp filed Critical NEC Corp
Priority to EP20917095.0A priority Critical patent/EP4098736A1/en
Priority to PCT/JP2020/003466 priority patent/WO2021152780A1/ja
Priority to US17/794,027 priority patent/US20230133438A1/en
Priority to JP2021574370A priority patent/JPWO2021152780A1/ja
Publication of WO2021152780A1 publication Critical patent/WO2021152780A1/ja
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor

Definitions

  • the present invention relates to a DNA extraction method and a nail processing device.
  • an object of the present invention is to provide a DNA extraction method and a nail processing apparatus that contribute to improving the efficiency of DNA extraction from nails.
  • a processing step of processing a nail as a sample for DNA extraction while maintaining the state of one mass to increase the surface area of the nail exposed to the DNA extraction reagent and a processing step.
  • An extraction step of exposing the processed nail to a DNA extraction reagent to extract DNA, A DNA extraction method comprising the above is provided.
  • a nail processing device that processes a nail as a sample for DNA extraction while maintaining the state of one mass to increase the surface area of the nail exposed to the DNA extraction reagent. Provided.
  • a DNA extraction method and a nail processing device that contribute to improving the efficiency of DNA extraction from nails are provided.
  • the DNA extraction method of the present invention processes a nail as a sample for DNA extraction while maintaining the state of one mass to increase the surface area of the nail exposed to the DNA extraction reagent.
  • the machining step (step S01) is included.
  • the DNA extraction method of the present invention includes an extraction step (step S02) in which the processed nail is exposed to a DNA extraction reagent to extract DNA.
  • the nail as a sample for DNA extraction is rolled flat by the nail processing device.
  • the nail has an appropriate stickiness in the first place, and the nail can maintain the state of one lump even in the state of being rolled into a flat shape. That is, by processing the nail into a flat shape, the surface area of the nail can be increased while maintaining the state of one mass.
  • the flattened nail is transferred to a container for DNA extraction (for example, a microtube) and exposed to a DNA extraction reagent to extract DNA.
  • a container for DNA extraction for example, a microtube
  • the nail transferred into the container has a frictional force with the inner wall of the container for DNA extraction, the nail does not float on the DNA extraction reagent. Further, even if it floats, the nail can be settled in the DNA extraction reagent much more easily than the shattered nail, for example, by pushing it into the DNA extraction reagent with a rod-shaped instrument.
  • the DNA extraction efficiency from the nail can be improved. That is, since the nail is processed while maintaining the state of one lump, it can be easily taken out from the processing apparatus without loss. In other words, the processing equipment is easier to clean and the risk of carryover / contamination with the next sample is reduced. In addition, since the nail remains in the state of one mass, it is easy to settle in the DNA extraction reagent.
  • the nail processing device has a basic property of processing a nail as a sample for DNA extraction while maintaining the state of one mass to increase the surface area of the nail exposed to the DNA extraction reagent. In other words, the nail processing device only needs to be able to process the nail so as to increase the surface area while maintaining the state of one mass.
  • the claw processing device is exemplified by a press machine in which a claw is sandwiched between a pair of dies and pressure is applied to roll the claw into a flat shape.
  • the pressure applied to the nail is preferably about 200 to 2000 kgf (about 2000 to 20000 N (Newton)), but may be 200 kgf or less, or 2000 kgf or more. That is, if the pressure applied to the nail is large, the surface area of the flatly rolled nail is also large, but the thickness of the nail is thin and it is difficult to maintain the state of one lump, which is a disadvantage. Occurs.
  • the pressure applied to the nail can be selected in consideration of various factors such as the skill of the operator and the purpose of DNA extraction.
  • the mechanism for applying pressure to the claw can be selected from a wide variety of mechanisms such as a human-powered mechanism (for example, a mechanism using a hydraulic jack) and a mechanical mechanism.
  • the material and shape of the mold can be changed in a wide variety of ways if the nails can be processed so as to increase the surface area while maintaining the state of one lump.
  • One example is a die made of stainless steel and having a flat press surface.
  • the pressed surface can be provided with irregularities with the intention of further increasing the surface area of the nail.
  • the nails may be stretched by performing operations such as relatively moving or rotating the pair of molds.
  • the press surface of the die may be provided with an inclination.
  • the pressure applied to the claws, the movement of the mold, and the rotation may be performed at the same time or separately.
  • the nail can be processed so as to increase the surface area while maintaining the state of one mass, but even if the location of the nail to which pressure is applied is gradually changed. good.
  • the pressure may be applied to the central portion of the nail first, and then the portion of the nail to which the pressure is applied may be gradually shifted outward. In this case, since the nail is gradually pushed outward from the central portion, more efficient rolling of the nail is possible.
  • the claw processing device is also exemplified by a roller mechanism in which a claw is sandwiched between a pair of rollers and the claw is rolled into a flat shape.
  • spike rows (s) along the circumference may be provided on the surface of the roller.
  • the rolling direction of the claw is restricted to the rolling direction of the rollers (the direction of insertion between the rollers), and the claws are rolled in the axial direction.
  • the rolling of the claws can be restricted. This processes the claws into a sheet of appropriately connected strips, which makes it easier to maintain the state of a single mass (for example, imagine a squid).
  • the spike rows may be arranged not only along the circumference of the roller but also diagonally along the circumference of the roller.
  • the claw processing device is also exemplified by a hammer mechanism that strikes a claw with a hammer to stretch the claw.
  • FIG. 2A shows an example of a press machine as a claw processing device (a hydraulic experimental pellet press manufactured by Specac).
  • the die used has a flat press surface.
  • a pressure of about 2000 kgf is applied to the nail, the nail is processed into an elliptical sheet having a major axis of 7 mm and a minor axis of about 5 mm as shown in FIG. 2B.
  • the nail has a hardness enough to be pinched with tweezers and a softness enough to be flexibly bent, so that it can be easily peeled off from the mold.
  • a cartridge for DNA analysis for example, RapidHIT manufactured by Thermo Fisher Scientific Co., Ltd.
  • RapidHIT manufactured by Thermo Fisher Scientific Co., Ltd.
  • This cartridge is originally designed so that a swab with oral cells can be inserted, and when inserting it into the cartridge, the nail is wrapped around a rod-shaped instrument, cut into multiple strips, and so on. Alternatively, it may be pushed in forcibly.
  • the subsequent treatment to dissolve the nails is generally established as an experimental system, so explanations will be omitted. That is, in the present invention, the process of applying the processed nail instead of the oral cells attached to the swab is a technical difference from the general DNA analysis method.
  • a jig having a handle may be provided to process the nail so as to be integrated with the jig.
  • the holding tool having the handle portion has a frame portion designed to be along the outer circumference of the press surface, for example, as shown in FIG.
  • a part of the frame portion may be crushed by press working.
  • the frame portion is designed so as to be crushed and extended inward, the contact between the claw and the frame portion becomes easier.
  • FIG. 5 it may have a whiskers portion extending inward from the frame portion and pressed together with the claws.
  • the whiskers can be spiral as shown in FIG. 5A or radial as shown in FIG. 5B.
  • a binder may be further provided, and the holding tool and the claw may be integrally processed via the binder.
  • the binder is preferably cotton-like or mesh-like in consideration of exposure of the nail to the DNA extraction reagent.
  • the material of the binder may include a natural material such as cotton, a plastic resin, plastic and the like.
  • the sample can be applied not only to nails but also to bones and teeth. Since bones, teeth, etc. may be crushed into pieces when pressed, it is preferable to press them together with a thickener for maintaining the state of one mass.
  • Appendix 2 The DNA extraction method according to Appendix 1, wherein the nail is processed into a flat shape in the processing step.
  • Appendix 3 The DNA extraction method according to Appendix 2, wherein the nail is press-processed in the processing step.
  • Appendix 4 The DNA extraction method according to Appendix 3, wherein in the processing step, a holding tool having a handle is provided and the nail is processed so as to be integrated with the holding tool.
  • a nail processing device that processes a nail as a sample for DNA extraction while maintaining the state of one mass to increase the surface area of the nail exposed to the DNA extraction reagent.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Plant Pathology (AREA)
  • Biophysics (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Sampling And Sample Adjustment (AREA)
PCT/JP2020/003466 2020-01-30 2020-01-30 Ｄｎａ抽出方法及び爪加工装置 Ceased WO2021152780A1 (ja)

Priority Applications (4)

Application Number Priority Date Filing Date Title
EP20917095.0A EP4098736A1 (en) 2020-01-30 2020-01-30 Dna extraction method and nail processor
PCT/JP2020/003466 WO2021152780A1 (ja) 2020-01-30 2020-01-30 Ｄｎａ抽出方法及び爪加工装置
US17/794,027 US20230133438A1 (en) 2020-01-30 2020-01-30 Dna extraction method and nail processor
JP2021574370A JPWO2021152780A1 (https=) 2020-01-30 2020-01-30

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/JP2020/003466 WO2021152780A1 (ja) 2020-01-30 2020-01-30 Ｄｎａ抽出方法及び爪加工装置

Publications (1)

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WO2021152780A1 true WO2021152780A1 (ja) 2021-08-05

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US (1) US20230133438A1 (https=)
EP (1) EP4098736A1 (https=)
JP (1) JPWO2021152780A1 (https=)
WO (1) WO2021152780A1 (https=)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI862371B (zh) * 2023-12-29 2024-11-11 致茂電子股份有限公司 用於萃取生物檢體之攪拌棒

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2004201558A (ja) * 2002-12-25 2004-07-22 Wako Pure Chem Ind Ltd Ｄｎａ遊離方法
JP2009011210A (ja) 2007-07-03 2009-01-22 Keio Gijuku Ｄｎａ抽出方法及びそのためのキット
JP2014113079A (ja) * 2012-12-07 2014-06-26 Mitsubishi Chemical Medience Corp 高精度な白癬菌の検出法
JP2017518497A (ja) * 2014-05-30 2017-07-06 プレッシャー バイオサイエンシズ インコーポレイテッドＰｒｅｓｓｕｒｅ Ｂｉｏｓｃｉｅｎｃｅｓ，Ｉｎｃ． 試料調製デバイスおよび方法

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10429279B2 (en) * 2013-07-28 2019-10-01 Xmd, Llc Devices and cartridges for extracting bio-sample regions and molecules of interest
EP3179912B1 (en) * 2014-08-15 2020-10-14 Diomics Corporation Films for biologic analyte collection and analysis and methods of production

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2004201558A (ja) * 2002-12-25 2004-07-22 Wako Pure Chem Ind Ltd Ｄｎａ遊離方法
JP2009011210A (ja) 2007-07-03 2009-01-22 Keio Gijuku Ｄｎａ抽出方法及びそのためのキット
JP2014113079A (ja) * 2012-12-07 2014-06-26 Mitsubishi Chemical Medience Corp 高精度な白癬菌の検出法
JP2017518497A (ja) * 2014-05-30 2017-07-06 プレッシャー バイオサイエンシズ インコーポレイテッドＰｒｅｓｓｕｒｅ Ｂｉｏｓｃｉｅｎｃｅｓ，Ｉｎｃ． 試料調製デバイスおよび方法

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
FENG TAO, CHUNQIN LI, GARY SMEJKAL, ALEXANDER LAZAREV, NATHAN LAWRENCE, RICHARD T SCHUMACHER: "Pressure Cycling Technology (PCT) Applications in Extraction of Biomolecules from Challenging Biological Samples", HIGH PRESSURE BIOSCIENCE AND BIOTECHNOLOGY 166 PROCEEDINGS OF THE 4 TH INTERNATIONAL CONFERENCE ON HIGH PRESSURE BIOSCIENCE AND BIOTECHNOLOGY, 1 January 2007 (2007-01-01), pages 166 - 173, XP055458730, Retrieved from the Internet <URL:http://www.exaltedweb.com/pressure/file_579f69f1aa03c_HPBB-proceeding-paper.pdf> *
HASHIYADA, M. ; NAKAYASHIKI, N. ; FUNAYAMA, M.: "Utility validation of extraction of genomic DNA from hard tissues, bone and nail, using PrepFiler(TM) Forensic DNA Extraction Kit", FORENSIC SCIENCE INTERNATIONAL: GENETICS SUPPLEMENT SERIES, ELSEVIER IRELAND LTD, AMSTERDAM, NL, vol. 2, no. 1, 1 December 2009 (2009-12-01), AMSTERDAM, NL, pages 172 - 173, XP026791816, ISSN: 1875-1768 *
KOTROTSIOU T., CHATZIMICHALOGLOU A., NASR A., SPANDOU E., PATSATSI A., SOTIRIADIS D., VYZANTIADIS T.-A.: "A comparative study of three extraction protocols of DNA from nails: Potential use in the diagnosis of onychomycoses", MYCOSES, WILEY-BLACKWELL PUBLISHING LTD., GB, vol. 60, no. 3, 1 March 2017 (2017-03-01), GB, pages 183 - 187, XP055844268, ISSN: 0933-7407, DOI: 10.1111/myc.12581 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI862371B (zh) * 2023-12-29 2024-11-11 致茂電子股份有限公司 用於萃取生物檢體之攪拌棒

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US20230133438A1 (en) 2023-05-04
EP4098736A1 (en) 2022-12-07

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