WO2018053191A1 - Appareil et procédé pour collecter de l'adn de contact - Google Patents

Appareil et procédé pour collecter de l'adn de contact Download PDF

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Publication number
WO2018053191A1
WO2018053191A1 PCT/US2017/051650 US2017051650W WO2018053191A1 WO 2018053191 A1 WO2018053191 A1 WO 2018053191A1 US 2017051650 W US2017051650 W US 2017051650W WO 2018053191 A1 WO2018053191 A1 WO 2018053191A1
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WO
WIPO (PCT)
Prior art keywords
collector
dna
nucleic acids
collection
chemical composition
Prior art date
Application number
PCT/US2017/051650
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English (en)
Inventor
Irina KIRGIZ
Cassandra CALLOWAY
Original Assignee
The Regents Of The University Of California
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 The Regents Of The University Of California filed Critical The Regents Of The University Of California
Publication of WO2018053191A1 publication Critical patent/WO2018053191A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/02—Devices for withdrawing samples
    • 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
    • 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
    • C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
    • 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
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/02—Devices for withdrawing samples
    • G01N2001/028—Sampling from a surface, swabbing, vaporising

Definitions

  • the technology of this disclosure pertains generally to forensic analysis
  • nucleic acid material collection devices and fabrication methods and more particularly to a device for collecting, purifying, and storing nucleic acids from low copy DNA sources for later identification.
  • DNA is the genetic blueprint of an individual and each individual's DNA is unique. Like fingerprints, the analysis of DNA is a highly
  • DNA samples are typically collected from blood, saliva, semen, and other bodily fluids as well as from bones, hair and skin found at a crime scene.
  • DNA evidence is collected at a crime scene to accurately detect and identify suspects as well as rule out suspects and to establish other facts related to the crime.
  • DNA crime scene evidence may be presented at trial to assist a jury in their determination of the guilt or innocence of the accused.
  • PCR Polymerase chain reaction
  • the small DNA sample is used as a replication template for creating large numbers of copies that can be easily detected and evaluated by conventional DNA analysis techniques.
  • the effectiveness and reproducibility of the PCR amplification is dependent may be greatly influenced by the purity and quantity of the sample DNA template. Samples may not only have contaminating DNA but they may also have molecules that inhibit PCR amplification or damage the sample. Therefore, precise collection and the removal or deactivation of inhibitors is important to performing successful PCR amplifications.
  • DNA fingerprints from fingerprints is called “Touch DNA” and typically yields very small amounts of low copy number (LCN) DNA, often from only one or two cells.
  • Touch DNA may be present in limited quantities at a crime scene, and high recovery is critical in order to allow DNA typing. In many cases, touch DNA samples only yield a partial profile that is frequently attributed to the low shedding status of a donor. Low yields can also be explained by ineffective collection methods that leave a portion of deposited DNA behind.
  • touch DNA evidence Forensic analysis of touch DNA evidence was first described in 1997 by Van Oorschot and Jones. Since then, the topic of touch DNA has become of particular interest to researchers in the field of Forensic Science. Now this type of evidence is frequently used in criminal cases worldwide and the number of cases solved solely by virtue of touch DNA has grown dramatically. Each year approximately 800 thousand vehicles are stolen in the United States and an estimated 4.9 million vehicles are stolen worldwide. Many of the stolen vehicles are used to transport illegal substances or are involved in other types of crimes. In many cases, touch DNA collected from the steering wheel is the only evidence that can link a perpetrator to the crime (e.g. most recent driver to the carjacking).
  • touch DNA Despite the widespread use of touch DNA evidence, there is no standardized touch DNA collection protocol. The collection methods used for low copy DNA specimens by many labs were optimized for high copy number DNA evidence. Not all techniques that are used for blood, sperm and saliva DNA collection are effective for collecting touch DNA. Since most of the time bodily fluids are present in abundance at the crime scene, there is no need to collect DNA from the entire specimen. Touch DNA, on the other hand, is present in limited quantities and therefore high recovery is critical.
  • Collection swabs are typically kept in sterile containers or bags before use and returned to the sterile container after collection to limit contamination.
  • the container is usually marked with information such as the time, date, location and the identity of the collector to establish a chain of custody for the sample.
  • Conventional touch DNA collection devices are designed to collect touch DNA over a small surface area.
  • the swab for example, has a small surface area that becomes saturated with extraneous material very quickly. Once saturated, the swab can smear or spread additional and already- collected material over a large area rather than collecting more material.
  • the location of the deposited touch DNA is typically unknown and can only be approximated. Therefore, there is a need for a method that can effectively collect small deposits of touch DNA scattered over a large surface area.
  • a DNA collection device for efficient and reliable recovery of forensic low copy number DNA from fingerprints.
  • the present technology generally provides a biological material collector apparatus that is designed to speed up and increase the efficiency of the touch DNA collection process and similar acquisitions of low copy DNA.
  • the device is a sensitive crime scene collection tool for maximum recovery of trace forensic DNA evidence that has been left at a crime scene for subsequent DNA PCR analysis.
  • the apparatus provides for the efficient collection, storage, and
  • nucleic acids such as DNA or RNA
  • the nucleic acids can be released from the collector after collection or storage and then amplified by conventional amplification methods such as polymerase chain reaction and evaluated.
  • DNA analysis of collected trace DNA can be obtained by generating small tandem repeat (SIR) profiles,
  • the collector has a preferably flexible base with an optional handle or coupling for reversibly attaching a handle to the base.
  • the base of the collector may be made from a flexible foam square or may be formed from a rigid material.
  • a collector matrix sheet with a liquid impermeable membrane on the back side may be mounted to the base or directly to the impermeable membrane.
  • the backing membrane may comprise cellophane or other material that prevents liquid from permeating through.
  • the collector matrix and backing membrane are not joined with a base.
  • the collector has a larger surface area than double swabs and can cover larger surface areas before drying out and/or saturating with a material, when compared to traditional touch DNA collection methods (e.g., double swabbing).
  • the collector matrix layer is preferably an absorbent layer that can be made from a variety of materials that may be woven fabrics, nonwoven materials or foams.
  • Preferred materials include such as cotton, nylon, cellulose-based materials such as blotter or filter paper as well as paper/cloth blend materials and microcellular foams and similar materials. Preferred materials do not bind nucleic acids irreversibly.
  • the absorbent collector matrix is preferably impregnated with a
  • the chemical composition that lyses cells and denatures proteins and separates the nucleic acids. When cells, for example, are collected by the collection matrix they are lysed and the nucleic acids are immobilized and stabilized within the collector matrix. The DNA is effectively deposited directly on to the coSiector matrix eliminating the need to transfer the DNA to a separate storage material that often decreases the overall DNA yield.
  • the chemical composition can include components that can inhibit microbial growth that can damage the DNA and pollute the identification schemes.
  • the chemical composition comprises a mixture of a weak base; a chelating agent; an anionic detergent; and optionally a uric acid or urate salt. Particularly preferred embodiments include a weak base of tris(hydroxymethyl)methane, a chelating agent comprising
  • EDTA ethylenediaminetetraacetic acid
  • SDS sodium dodecyl sulfate
  • FIG. 1 is a schematic exploded side view of a collector according to one embodiment of the technology.
  • FIG. 2A is a front view of a steering wheel structure and sampling of water-soluble tape lifting or double swabbing collection techniques that were used to collect touch DNA from the surface of the steering wheel and then compared.
  • FIG. 2B is a front view of a steering wheel structure and sampling of FTA paper scraping or double swabbing collection techniques that were used to collect touch DNA from the surface of the steering wheel and compared.
  • FIG. 3 is a graph of% STR Allele Recovery of Last Driver's Profile for the steering wheels of eight cars that were compared based on how well the swab or tape collection technique recovered the last driver's profile.
  • FIG. 4 is a graph of% STR Allele Recovery of Last Driver's Profile for the steering wheels of eight cars that were compared based on how well the swab or FTA pad collection technique recovered the last driver's profile.
  • FIG. 5 is a functional block diagram a method for using the collector apparatus for the collection and storage of low copy number DNA and analysis for forensic investigation and evidence.
  • FIG. 1 and FIG. 5 illustrate the characteristics and functionality of the apparatus and system. It will be appreciated that the methods may vary as to the specific steps and sequence and the systems and apparatus may vary as to structural details without departing from the basic concepts as disclosed herein. The method steps are merely exemplary of the order that these steps may occur. The steps may occur in any order that is desired, such that it still performs the goals of the claimed technology.
  • the apparatus for low copy nucleic acid collection is
  • FIG. 1 one embodiment of a collector 10 that is
  • the collector 10 shown in the exploded view of FIG. 1 has a collector sheet 12 that is mounted to a liquid impermeable membrane 14.
  • the membrane 14 is coupled to a base 16 that has a handle 18 to allow the user to avoid touching and contaminating the collector sheet 12 during use.
  • the handle 18 can be a hard plastic member such as a disk that is mounted with a generally perpendicular orientation to the face 20 of the collector sheet 12. The user can pinch the handle 18 with the first finger and thumb during use.
  • the handle 18 is a shaft of a variety of lengths that further isolates the user from the collection surface 20 of collector sheet 12 during use.
  • the handle 18 is a loop of flexible material that can receive one or two fingers therein to facilitate use.
  • the base 16 has a coupling that permits the collector 10 to reversibly couple the base 16 to a telescoping handle 18, for example.
  • the base 16 is preferably made from a flexible and resilient material such as foam to allow flexibility in the collector to adapt to rough or non- planer surfaces including cylindrical or rounded surfaces.
  • the base 16 is made from a rigid material making the collector 10 particularly suited for collections from smooth planar surfaces. Samples are typically collected from surfaces such as metal, plastic, concrete, vinyl, wood, glass, and even some fabrics.
  • the membrane 14 separating the collector sheet 12 from the base 16 is preferably liquid impermeable so that biological material that is collected by the collector sheet 12 is isolated and contained by the collector sheet 12.
  • the membrane 14 is made from a non- reactive and non-porous material such as cellophane.
  • the membrane is made from a hydrophobic material. The membrane 14 prevents liquid from permeating through to the base and increases the DNA collection yield by preventing loss of collected DNA material from the collector sheet 12.
  • an apparatus for collecting touch DNA from a surface comprises an "collector matrix" (a matrix comprising cotton, a paper/cloth blend material or any type of cellulose-based material, or filter material preferably contacted with a chemical composition that lyses cells and denatures proteins), and one or more sides of the collector matrix are contacted with a non-reactive, non-porous backing membrane 14.
  • collector matrix a matrix comprising cotton, a paper/cloth blend material or any type of cellulose-based material, or filter material preferably contacted with a chemical composition that lyses cells and denatures proteins
  • the collector sheet 12 is preferably absorbent and porous and
  • Suitable materials for the collector sheet 12 should not irreversibly bind nucleic acids (i.e. through covalent or ionic bond formation) and preferably should be substantially hydrophilic or capable of being made hydrophilic.
  • the porous material of the collector sheet 12 may be a paper/cloth blend, cotton fibers, filter material, blotter paper, absorbent foam or a cellulose-based material, for example.
  • the porous material of the collector sheet 12 may also be woven or non-woven or be made of porous polymer based materials such as nylon or polyester etc.
  • the chemical composition is composed of a weak base; a chelating agent an anionic detergent; and optionally a uric acid or urate salt.
  • the weak base is tris(hydroxymethyl)methane (THYM) or Tris(hydroxymethyl)aminomethane
  • the chelating agent is ethylenediaminetetraacetic acid (EDTA)
  • the detergent is sodium dodecyl sulfate (SDS).
  • the surface 20 of collecting sheet 12 may be primed with an optional collection solution such as a few drops (1 -8) of deionized water or other solvent to dampen the collector sheet 12.
  • an optional collection solution such as a few drops (1 -8) of deionized water or other solvent to dampen the collector sheet 12.
  • Buffered saline, lysis buffer, SDS are suitable alternatives to water.
  • a dampened collection sheet 12 increased the efficiency of the collector in the acquisition of small numbers of cells and/or amounts of nucleic acids.
  • the collectors 10 may be kept in sterile containers that can be
  • the containers can be marked with sample collection information such as the date, time and place of the sample collection as well as the name of the person taking the sample.
  • the stabilized nucleic acids in the absorbent collector sheet 12 may be stored with the collector for a period of time before processing or the collecting sheet 12 may be separated and removed from the collector body and stored separately. In one embodiment, only a portion of the collecting sheet 12 is processed and a second portion is stored for archival purposes and later processing if there is some question about the results from the first portion.
  • Collectors 10 and containers may be individually packaged or they may be assembled into self-contained sampling kits.
  • Sampling kits may include instructions, labels, log, collection solution, and a number of collectors and storage containers, for example.
  • the kit can be used to safely transport the collected samples without leakage from the site for further analysis.
  • the sealed container of the kit that stores the used collector can also serve as a long term storage container.
  • the collectors 10 can be used with high sensitivity PCR DNA testing protocols for amplification.
  • the collectors can also be used with a method for collecting and preserving nucleic acids from trace forensic biological samples in one step because the collection matrix also serves as a sample preservation and storage matrix.
  • the collectors provide a method of acquiring and analyzing trace DNA from a fingerprint or trace cells. Referring now to FIG. 5, one method 100 for the collection and identification of forensic touch DNA is shown.
  • low copy number (LCN) DNA is acquired in an analyzable amount with the use of the collector apparatus and placed and sealed in a container.
  • the sample that has been stabilized the DNA in the collector and sealed in the container can be transported and preserved without contamination.
  • the stabilized DNA in the collector and container can be stored for long periods of time if necessary before analysis.
  • the sample can also be divided and portions analyzed and other portions stored to replicate results if necessary.
  • the trace DNA is released from the collector for analysis.
  • the DNA samples may be further purified from the cell constituents upon release. Because of the small quantities of nucleic acids that are present in the sample, the released DNA is preferably amplified and analyzing using a high sensitivity PCR DNA testing protocol in the step at block 140 of FIG. 5.
  • the amplified DNA is now in sufficient quantities to reliably analyze unique markers and other characteristics of the DNA to identify its origins. Accordingly, at block 150, multiple loci of the DNA are identified for comparison with DNA marker entries in a crime database or to processed DNA samples from a specific suspect.
  • collectors similar to that shown in FIG. 1 were prepared and evaluated.
  • the collector had a collector sheet of a 3 cm x 3 cm cotton fabric that had been soaked in a mixture of chemicals that lyses cells and denatures proteins.
  • One side of the collector fabric was covered with a non-reactive, non-porous cellophane backing that prevented liquid from permeating through the collector fabric.
  • the collector fabric was then attached to a foam base of a 2.8 cm x 2.8 cm flexible foam square that contained a handle located in the middle of the square for a better grip.
  • the prepared collector fabric was connected to the applicator and 4 drops of deionized water was administered to the surface of the fabric. Then, the collector was used to scrape or rub the surface of the fabric against the surface of the steering wheel 22 in a circular motion as shown in FIG. 2B. Fingers could be used to apply light pressure to the foam part of the collector for better collection results or to go over curvy areas.
  • the collector described herein has a larger surface area and can cover larger surface areas before drying out and/or saturating with a material, when compared to traditional touch DNA collection methods (e.g., double swabbing).
  • the collector has all the advantages of FTA cards (no extraction process, long term DNA storage), plus it allows one to deposit a DNA directly on an storage fabric that eliminates the need to transfer DNA that in turn can decrease the overall DNA yield.
  • the collector in this example used a modified version of FTA paper (wet FTA paper easily tears appart on a rough surface); that is, filter paper has been replaced by a cotton fabric that is firmer and does not break appart easily. It was found that FTA paper collected higher yields of touch DNA compared to standard DNA collection method.
  • one half of a steering wheel 22 was sampled using a double swabbing technique with two swabs 24 and the other side of a steering wheel 22 was sampled tape 26 applied over the surface of the wheel.
  • one half of a steering wheel 22 was sampled using a double swabbing technique with swabs 28, while the other side of a steering wheel was sampled using the FTA paper scraping technique with the collector 30 as shown in FIG. 2B.
  • Swabs 24, 28 in both evaluations were moistened with de-ionized water before the sample collection.
  • Each Whatman WB120205 FTA Classic Card was cut into four pieces; each piece was used to scrape one side of a steering wheel. Four drops of de- ionized water were applied to an FTA card before the sample collection.
  • a 6 cm piece of 3MTM Water-Soluble Wave Solder Tape 5414 was used to collect DNA from one side of the steering wheel.
  • DNA was thereafter extracted from the collected samples from the swabs, buccal swabs, collector and tape using commercial DNA extraction kits. Then, real-time qPCR analysis using the Promega® PlexorHY Human Quantitation kit following the manufacturer's procedure was used to determine the amount of DNA present in each sample. Each sample was quantified two times to increase the accuracy of the estimated DNA amount; if a CT difference greater than 0.5 was observed between the replicates then the quantitative analysis was repeated.
  • Detectable quantities of touch DNA were recovered from each of the 70 steering wheels sampled independent of collection method and randomized for collecting from the right or the left side for a total of 140 samples.
  • the FTA paper scraping method was shown to be the most effective for collecting DNA from steering wheels followed by double swabbing and tape lifting methods.
  • the double swabbing method collected slightly higher amounts of touch DNA compared to tape lifting; however, the difference was not statistically significant.
  • DNA yields using the double swabbing collection method were most highly dispersed, ranging from 0.08 ng to 883.75 ng.
  • the swab has a relatively small surface area and it is a very tedious and time consuming process to collect from the entire surface of the steering wheel with a swab. Since there was no visual indication as to whether or not the area was covered by swabbing, certain areas could have been covered twice while others missed. Using a second wet swab immediately following the first one increased the DNA yield and was prone to the same limitations as the first swab collection.
  • FTA paper collect more touch DNA from the surface of a steering wheel as compared to the double swabbing method.
  • the FTA paper was larger when compared to swabs and could cover a greater surface area before getting saturated.
  • the FTA paper did not dry as fast as swabs and still appeared wet at the end of the collection process.
  • the effectiveness of FTA paper remained approximately the same throughout the entire collection process, which allowed it to cover large surface areas in one attempt. Unfortunately, wet FTA paper easily tears apart on rough surfaces which may have
  • FTA paper still can be used to collect DNA from flat and smooth surfaces such as mirrors, table tops, shelves and floors. This method could be useful in situations when the exact location of touch DNA is unknown or small quantities of touch DNA are dispersed throughout the surface.
  • the goal of the STR analysis was to assess the difference between quantifiable and typable results among the three collection methods.
  • the results show that the double swabbing method is more likely to recover the last driver's profile when compared to the tape lifting method and that the FTA paper scraping method recovers similar completeness of the profile as the double swabbing method (FIG. 3-4).
  • present disclosure encompasses multiple embodiments which include, but are not limited to, the following:
  • the method comprising: (a) contacting a surface with a collector to acquire trace nucleic acids, the collector comprising: (i) a collection matrix; (ii) a chemical composition that lyses cells and denatures proteins wherein the chemical composition is contacted to the collection matrix; and (iii) a non-porous, non-reactive backing wherein the backing is contacted with one or more surfaces of the collection matrix; (iv) wherein trace nucleic acids are freed from cells or organisms on contact with the chemical composition; and (b) preserving the acquired trace nucleic acids in the collection matrix of the collector for a period of time prior to analysis.
  • the collection matrix is a material selected from the group of materials consisting of blotting paper, filter paper, cotton fibers, nylon fibers, polyester fibers, and a blend of paper and cloth fibers.
  • the chemical composition comprises a mixture of a weak base; a chelating agent; and an anionic detergent.
  • chemical composition further comprising: a uric acid or a urate salt.
  • a method for identifying nucleic acid origins from trace forensic biological samples comprising: (a) contacting a surface with a collector to acquire trace nucleic acids, the collector comprising: (i) a collection matrix with an inert, impermeable backing layer with an inner surface and an outer surface and an absorbent layer mounted to the inner surface of the backing layer; and (ii) a chemical composition that lyses cells and denatures proteins-contacted to the absorbent layer of the collection matrix; (iii) wherein trace nucleic acids are freed from cells or organisms on contact with the chemical composition; (b) releasing acquired trace nucleic acids from the absorbent layer of the collection matrix of the collector; (c) amplifying purified released nucleic acids; (d) analyzing amplified nucleic acids for identifiable markers; and (e) comparing identified nucleic acid markers with nucleic acids from a specific suspect or from a crime database of nucleic acids to identify a match.
  • An apparatus for collection of nucleic acids from a surface comprising: (a) an inert, impermeable backing layer with an inner surface and an outer surface; (b) an absorbent layer mounted to the inner surface of the backing layer; and (c) a chemical composition capable of lysing cells and denaturing proteins disposed on a collecting surface of the absorbent layer; (d) wherein cells and proteins of biological samples collected at the collecting surface of the absorbent layer are lysed and denatured by the chemical composition; and (e) wherein nucleic acids from the collected samples are absorbed by the absorbent layer remain intact.
  • planar base mounted to the impermeable backing layer; and at least one handle coupled to the planar base.
  • the absorbent layer is at least one material selected from the group of materials consisting of a cotton material, a paper/cloth blend material, a cellulose-based material and a filter material.
  • the absorbent layer of the collection matrix is a woven material selected from the group of materials consisting of nylon, polyester, cotton, cellulose, and a blend of paper and cloth fibers.
  • the absorbent layer comprises an absorbent foam.
  • the backing layer comprises cellophane.
  • the chemical composition comprises: (a) a weak base; (b) a chelating agent; (c) an anionic detergent; and (d) a uric acid or urate salt.
  • the weak base comprises tris(hydroxymethyl)methane.
  • the chelating agent comprises ethylenediaminetetraacetic acid (EDTA).
  • EDTA ethylenediaminetetraacetic acid
  • the detergent comprises sodium dodecyl sulfate (SDS).

Abstract

L'invention concerne des dispositifs et des procédés pour la collecte et le stockage d'ADN à faible nombre de copies tel qu'un ADN de contact pour une analyse ultérieure. Le collecteur a une matrice de collecteur comprenant un support non poreux et non réactif. La matrice est imprégnée d'une composition chimique capable de lyser des cellules et des protéines dénaturantes. Des acides nucléiques sont déposés directement sur la matrice de collecteur et stabilisés pour un stockage à long terme, et il n'est donc plus nécessaire de transférer l'ADN, étape qui peut diminuer le rendement d'ADN global. Le collecteur peut être utilisé pour collecter de l'ADN de contact sur de grandes surfaces avant le séchage ou avant de devenir saturé avec un matériau par comparaison avec les procédés classiques de collecte d'ADN de contact.
PCT/US2017/051650 2016-09-14 2017-09-14 Appareil et procédé pour collecter de l'adn de contact WO2018053191A1 (fr)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11656157B2 (en) * 2019-04-25 2023-05-23 Fremonta Corporation Powered sampling device

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140272983A1 (en) * 2013-03-15 2014-09-18 Ge Healthcare Uk Limited Solid matrix for the storage of biological samples
US20160047720A1 (en) * 2014-08-15 2016-02-18 Diomics Corporation Films for biologic analyte collection and analysis and methods of production and use thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20140272983A1 (en) * 2013-03-15 2014-09-18 Ge Healthcare Uk Limited Solid matrix for the storage of biological samples
US20160047720A1 (en) * 2014-08-15 2016-02-18 Diomics Corporation Films for biologic analyte collection and analysis and methods of production and use thereof

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11656157B2 (en) * 2019-04-25 2023-05-23 Fremonta Corporation Powered sampling device

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