EP0653920A1 - Procede d'obtention d'un echantillon cellulaire non liquide - Google Patents

Procede d'obtention d'un echantillon cellulaire non liquide

Info

Publication number
EP0653920A1
EP0653920A1 EP93919898A EP93919898A EP0653920A1 EP 0653920 A1 EP0653920 A1 EP 0653920A1 EP 93919898 A EP93919898 A EP 93919898A EP 93919898 A EP93919898 A EP 93919898A EP 0653920 A1 EP0653920 A1 EP 0653920A1
Authority
EP
European Patent Office
Prior art keywords
brush
cells
bristles
membrane
animal
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.)
Withdrawn
Application number
EP93919898A
Other languages
German (de)
English (en)
Inventor
Katherine W. Klinger
Brenda L. Richards
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.)
Genzyme Corp
Original Assignee
Genzyme 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 Genzyme Corp filed Critical Genzyme Corp
Publication of EP0653920A1 publication Critical patent/EP0653920A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B10/00—Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements
    • A61B10/02—Instruments for taking cell samples or for biopsy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B10/00—Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements
    • 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
    • C12Q1/24—Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B10/00—Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements
    • A61B10/0045—Devices for taking samples of body liquids
    • A61B10/0051—Devices for taking samples of body liquids for taking saliva or sputum samples
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B10/00—Other methods or instruments for diagnosis, e.g. instruments for taking a cell sample, for biopsy, for vaccination diagnosis; Sex determination; Ovulation-period determination; Throat striking implements
    • A61B10/02—Instruments for taking cell samples or for biopsy
    • A61B2010/0216—Sampling brushes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B17/00—Surgical instruments, devices or methods, e.g. tourniquets
    • A61B17/32—Surgical cutting instruments
    • A61B2017/320004—Surgical cutting instruments abrasive
    • A61B2017/320012—Brushes

Definitions

  • cervicovaginal lavage was found to be superior to scrape, swab or brush methods for obtaining cervicovaginal cells (Goldberg, GL et. al., Am. J. Obstet. Gynecol. 161 : 1169-72 (1989); Vermund, SH et. al., Ann. J. Obstet. Gynecol. 160: 304-8 (1989)).
  • cervicovaginal cells obtained by lavage are in a liquid format and therefore present the same problems with handling, processing and contamination as samples obtained by mouthwashing techniques.
  • DNA has been obtained from epithelial cells, spun from urine (Gasparinin, P. et. al., N. Engl. J. Med. 320: 809 (1989)); and from root and surrounding sheath cells obtained from plucked hairs (Higuchi, R. et. al.. Nature 332:543-6 (1988)).
  • samples so obtained often do not contain enough DNA for genetic analysis. Therefore hair and urine samples are not optimal for clinical analysis.
  • a simple, noninvasive technique for obtaining a non-liquid cell sample having an appropriate amount of genetic material for clinical analysis would be useful.
  • the invention relates to a simple, noninvasive process for obtaining a non- liquid cell sample from an animal for genetic analysis.
  • the process involves inserting an appropriately sized brush into an animal's oral cavity, contacting the brush with an oral membrane, agitating the brush against the membrane so that cells are captured within the bristles of the brush, removing the brush, which contains the captured cells and isolating the cells from the bristles of the brush for use in genetic analysis.
  • the brush is spiral, so that cells can be obtained by rotating the brush against an oral membrane.
  • Cells so obtained can then be analyzed using any of a number of known genetic techniques. For example, cells obtained according to the disclosed method can be lysed and the cell's DNA can be contacted with a suitable labelled probe. In addition, DNA so obtained can first be amplified to increase the amount of genetic material available for analysis.
  • a major advantage of using the process described herein for obtaining cells for genetic testing is that the non-liquid sample so obtained is easier to handle and to process than a liquid sample. Further a non-liquid sample is less prone to contamination (e.g by bacteria) and is more stable.
  • the invention relates to a process for obtaining a non-liquid cell sample from an animal (e.g. a human or a domesticated mammal) for genetic analysis.
  • the process employs a brush to obtain cells from within a animal's oral cavity.
  • Examples of cells which can be obtained from an animal's oral cavity include epithelial cells of the inner cheek or tongue.
  • a brush is inserted into an animal's oral cavity, contacted with an oral membrane and agitated (e.g. rotated or brushed) against the membrane for a sufficient period of time, so that cells are captured within the bristles of the brush.
  • the brush, which contains captured cells is then removed from the animal's mouth and cells are isolated from the bristles of the brush for use in genetic analysis.
  • a brush must be of an appropriate size to enable insertion into the animal's oral cavity.
  • the brush bristles should be free of potentially contaminating material.
  • a brush comprising mammalian hair should not be used in the subject method, because genetic material present within the hair could contaminate the sample and thereby interfere with the genetic analysis.
  • Preferred brushes for use in the subject invention have synthetic (e.g. nylon or dacron) bristles.
  • An especially preferred brush for use in the invention is the nylon bristle cytology brush, (Cytosoft Medical Packaging Corporation, Camarillo, CA), which is currently used for Pap Smear and Chiamydia testing.
  • a brush which contains captured oral cells can be analyzed directly or transported and/or stored for subsequent genetic analysis.
  • measures aimed at avoiding the dislodging of captured cells from a brush should be taken.
  • the brush can be placed in a container (e.g. an envelope) during transportation and/or storage.
  • Cells may be obtained from a brush by any method which avoids loss of captured cells.
  • a preferred method of obtaining cells from the brush is by contacting the cells with a solution.
  • the ceils can be obtained from the brush using a solution which lyses cells (e.g. sodium hydroxide, potassium hydroxide).
  • Genetic material from oral cells so obtained can then be analyzed genetically using any of a number of known genetic techniques, such as digestion with restriction endonucleases, ultraviolet light visualization of ethidium bromide stained agarose gels, DNA sequencing, or hybridization with detectable nucleic acid sequences (e.g. labelled probes).
  • the genetic material from cells obtained according to the disclosed method can be amplified using known amplification techniques, such as Q-beta-replicase and the poiymerase chain reaction (PCR) (Saiki, R.K. et. al.. Science 239:487-491 (1988); Wong, C, et. al.. Nature 330, 384-386 (1987) and U.S. Patent 4,683,202 to Muilis et. al.) to further increase the amount of genetic material available for genetic analysis.
  • known amplification techniques such as Q-beta-replicase and the poiymerase chain reaction (PCR) (Saiki, R.K. et
  • Example 1 Obtaining Cells From Oral Cavities Using either a Brush or a Wooden Spatula
  • the oral cavities of five human subjects were scraped with wooden spatulas for about 30 seconds, and in five human subjects, the CytosoftTM nylon bristle cytology brushes (Medical Packaging Corporation; Camarilo, CA) were rotated against the inner side of their cheeks for about 30 seconds.
  • the spatulas or brushes were soaked in test tubes containing 50mM NaOH for about 1 hour, after which one tenth volume of IM TRIS (pH8) was added to the test tubes to neutralize.
  • the cheek cell lysates obtained using the spatulas or the brushes were used directly in a poiymerase chain reaction (PCR) to amplify one or more of the following exons of the Cystic Fibrosis transmembrane regulator gene: exons 4, 10, 1 1 , 20 and 21.
  • Apropriate primers for the PCR were developed based on the nucleic acid sequence disclosed in Riordan, J.R. et al.. Science 245: 1066-1073 (1989).
  • PCR amplification reactions were performed using AmpliTaq DNA poiymerase and a DNA Thermal Cycler (Perkin Elmer). Cheek lysates were amplified for 28 cycles of 10 seconds at 94°, 10 seconds 55°C, and 30 seconds at

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • General Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Analytical Chemistry (AREA)
  • Pathology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Microbiology (AREA)
  • Immunology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biophysics (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Physics & Mathematics (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Medical Informatics (AREA)
  • Surgery (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

Procédé simple, non invasif d'obtention d'un échantillon cellulaire non liquide à l'aide d'une brosse, aux fins de l'analyse génétique.
EP93919898A 1992-08-07 1993-08-05 Procede d'obtention d'un echantillon cellulaire non liquide Withdrawn EP0653920A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US92987192A 1992-08-07 1992-08-07
US929871 1992-08-07
PCT/US1993/007382 WO1994003111A1 (fr) 1992-08-07 1993-08-05 Procede d'obtention d'un echantillon cellulaire non liquide

Publications (1)

Publication Number Publication Date
EP0653920A1 true EP0653920A1 (fr) 1995-05-24

Family

ID=25458604

Family Applications (1)

Application Number Title Priority Date Filing Date
EP93919898A Withdrawn EP0653920A1 (fr) 1992-08-07 1993-08-05 Procede d'obtention d'un echantillon cellulaire non liquide

Country Status (4)

Country Link
EP (1) EP0653920A1 (fr)
JP (1) JPH08501228A (fr)
CA (1) CA2137282A1 (fr)
WO (1) WO1994003111A1 (fr)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU1602295A (en) * 1994-01-14 1995-08-01 Jockey Club, The Non-invasive sampling method for nucleic acid analysis
EP0956083A4 (fr) * 1995-11-15 2000-07-05 Genelink Inc Dispositif non invasif d'identification
ATE472775T1 (de) * 1999-02-23 2010-07-15 Cdx Lab Inc Minimal-invasiver apparat und verfahren zum test zum auffinden von läsionen in der mundhöhle und in ähnlichem epithel
EP3669747B1 (fr) 2014-05-02 2022-03-30 United States Endoscopy Group, Inc. Dispositif de nettoyage pour un dispositif endoscopique
JP2016045103A (ja) * 2014-08-25 2016-04-04 日東電工株式会社 ガスクロマトグラフィー質量分析用試料サンプリング材
JP2016045102A (ja) * 2014-08-25 2016-04-04 日東電工株式会社 高速液体クロマトグラフィー用試料サンプリング材

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2839049A (en) * 1954-03-25 1958-06-17 Kenneth S Maclean Abrasive cytologic brush
GB2159420B (en) * 1984-05-29 1988-05-11 Medscand Ab Endocervical sampling brush and smear method
CA2071935C (fr) * 1990-10-26 1996-06-11 Arja Kallio Methode permettant d'evaluer la convenance des specimens cliniques pour les essais d'hybridation et trousse necessaire a l'valuation

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9403111A1 *

Also Published As

Publication number Publication date
CA2137282A1 (fr) 1994-02-17
WO1994003111A1 (fr) 1994-02-17
JPH08501228A (ja) 1996-02-13

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