CA2137282A1 - Process for obtaining a non-liquid cell sample - Google Patents
Process for obtaining a non-liquid cell sampleInfo
- Publication number
- CA2137282A1 CA2137282A1 CA002137282A CA2137282A CA2137282A1 CA 2137282 A1 CA2137282 A1 CA 2137282A1 CA 002137282 A CA002137282 A CA 002137282A CA 2137282 A CA2137282 A CA 2137282A CA 2137282 A1 CA2137282 A1 CA 2137282A1
- Authority
- CA
- Canada
- 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.)
- Abandoned
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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Pathology (AREA)
- Veterinary Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Heart & Thoracic Surgery (AREA)
- Medical Informatics (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- Public Health (AREA)
- Surgery (AREA)
- Microbiology (AREA)
- Animal Behavior & Ethology (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
A simple, non-invasive process for obtaining a non-liquid cell sample for genetic analysis using a brush is described.
Description
`-'?~'I/03~ 1;37~8~ PCl/US93/07382 .
PROCESS FOR BTAINING A NON-LIQUID CELL SAMPLE
, , Back~ound of the Imention Advances in recombinant DNA technology have made possible the molecular analysis and prediction of several genetic disorders. At the present time, ~enetic analyses are carried out mainly usin~ blood ieukocytes, which are 3 obtained from whole blood (e.g. via centrifu~ation). Whole blood, in turn must be obtained clinically by a physician or phlebotomist employing an invasive technique (e.g~ venipuncture).
i A non-invasive method for obtaining genetic material using a mouthwash has been describeci (Lench, N., et. al., The Lancet l: l356-8 (June l8, l988)). In this method, an individual rinses his mouth with saline which is then emitted from the mouth into o container, The container is centrifuged and cells, which are pelleted upon centrifugatlon, are collected. The ease with which sampies can be obtained is a major attraction of this method. However, samples so obtained are in a liquid format and therefore present probiems with regard to handling, 2 o processing and contamination.
DNA for genetic analysis has also been obtained from cervicovaginal cells ~Burk, R.D, ond C, Spitzer, Am. J. Obstet. GYnecol.l62:~52-4 (1990)). In astudy which compared the quantatlve yieid of cellular material obtained by 2 5 various methods, cervicovaginal lavage was found to be superior to scrape, swab or brush methocls for obtaining cervicovaginal cells (Goldberg, (~L et. al., Am. J. Obstet. GYnecol~ 16l: l 169-72 (1989); V~rmund, SH et. ai., Am. J. Obstet.
Gvnecol. 160: 304-8 (1989)). However, cervicovaginal cells obtained by lavage are in a liquid format and therefore present the same problems with handling, ; 1 ` ' 30 proaessing and contamination as scsmples obtained by mouthwashing techniques.
~: ' j;
WO 94/031 1 1 ~ 1 3 7 2 ~ 2 P~/US93/0738 '~=
For forensic purposes. DNA has been o~taine~ from epithelial cells, spun from urine (Gaspc~rinin, P. et. al., N. Enal. J. Med. 320: 80~ (1989)); and from root and surrounding sheath cells obtained from plucked hairs (Higuchi, R. et. al., Nature 332:543-6 (1988)). However, samples so obtained often do not contain S enough DNA for genetic analysis. Therelore hair and urine samples are not ' optimal for clinical ana~sis.
A simple, noninvasive technique for obtaining a non-liquid cell sample having an appropriate amount of genetic material for clinical analysis ~ould be 1 0 useful.
Summar,r o~ the InvenHon The invention relates to a simple, noninvasive process 'or obtaining a non-liquid cell sample from an animal for genetic analysis. The process involves inserting an appropriately srzed brush into an animal's oral cavily, contacting the brush with an oral membrane, agr~ating the brush against the membrane so that cells are captured within the bristles of the brush, removing the brush, which contains the captured ce!ls and isolating the cells from the bristles of the brush for use in genetic analysis. In a preferred embodiment, the brush is spiral, so thatcells can be obtained by rotating the brush against an oral mernbrane.
Cells so obtaineci can then be analyzed using any of a number of known genetic techniques. For example, cells obtained according to the disclosed method can be Iysed 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 arlalysis.
A major advantage of using the process described herein for obtaining ~ I 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 prane to contamination (e.g by bacteria~ and is more stable.
, J--`-'94~03ll~ 21~7X82 Pcr/US93/0738'`
l` ` '.
Detailed ~e~ ot the Invention In general, the invention relates to a process for obtainlng 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. Exarr ples of cells which can be obtained from an animal's oral cavity include epithelial cells of the inner cheek or tongue.
Accordin~ to the method of the invention, a brush is inserted into an anlmal'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.
To be suitable for use in the subject invention, a brush must be of an appropriate size to enable insertion into the animal's oral cavity. In addition, the brush bristles should be free of potentially contaminating material. For example, a brush comprising mammalian hair should not be used in the subject method, 2 0 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 docron) bristles. An especially preferr~d brush for use in ths invention is the nylon bristle cytology brush, (CytosottTM Medical Packaging Corporation, Camarillo, CA), which is currently 2 5 used for Pap Smear and Chlamydia testing.
Once removed from the mouth, a brush which contains captured oral cells can be analyzed directly or transported andlor stored for subsequent genetic analysis. At thls point, measures aimed at avoiding the dislodging of ~ I captured cells from a brush should be taken. For example, the brush can be pl~ced in a container (e.g. an envelope) during transportation and/or storage.
Cells may t~ obtained from a brush by any method which avoids loss of captured cells. A preferred method of obtaining cells from th t brush is by :
,:
,.~ , ~ ~ .
:`
WO 94/031 1 1 PCr/US93/0738'~
2137282 '~
contacting the cells with a solution. For example, if DNA is to be isolated from the cells for amplification, the cells c~n be obtained from the brush using a solution which Iyses cells (e.~. sodium hydroxide, potassium hydroxide).
Genetic material from oral celis so obtained can then be analyzed genetically using any of a number of known genetic techniques, such as di~estion with restriction endonucleases, ultraviolet light visualization of ethidium i~romld~ stalned a~arose g81s, DNA sequencing, or hybridization with detectable nucloic acid sequences ~e.~, labelled probes). In addi~ion, the genetic materialfrom cells obtalned accordins to the disclosed method can be amplified using known arnplification techniques, such as Q-beta-replicase and the polymerase chain reaction (PCR) (Saiki, R.K. et. al~, Science 239:487-491 (1~88); Wong, C., et.
al., Nature 330, 384-386 ~1987~ and U.S. Patent 4,683,202 to Mullis et. al.) to further increase the amount of genetic material available for genetic analysis.
The invention will now be illustrateci in the following Example, which is not intended to be limiting in any way.
Example 1: Obtaining Cells From Oral Cavities Using either a Brush or a Wooden 2 0 Spatula The oral cavities of five human subJects were scraped with wooden spatulas for about 30 seconds, and in five human subjects, the CytosottTM nylon bristle cytology brushes (Medical Packa~ing Corporation; Camarilo, CA) were 2 5 rotatad 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, a~ter whlch one tenth volume of 1 M TRIS (pH8) was added to the test tubes to neutralke.
PROCESS FOR BTAINING A NON-LIQUID CELL SAMPLE
, , Back~ound of the Imention Advances in recombinant DNA technology have made possible the molecular analysis and prediction of several genetic disorders. At the present time, ~enetic analyses are carried out mainly usin~ blood ieukocytes, which are 3 obtained from whole blood (e.g. via centrifu~ation). Whole blood, in turn must be obtained clinically by a physician or phlebotomist employing an invasive technique (e.g~ venipuncture).
i A non-invasive method for obtaining genetic material using a mouthwash has been describeci (Lench, N., et. al., The Lancet l: l356-8 (June l8, l988)). In this method, an individual rinses his mouth with saline which is then emitted from the mouth into o container, The container is centrifuged and cells, which are pelleted upon centrifugatlon, are collected. The ease with which sampies can be obtained is a major attraction of this method. However, samples so obtained are in a liquid format and therefore present probiems with regard to handling, 2 o processing and contamination.
DNA for genetic analysis has also been obtained from cervicovaginal cells ~Burk, R.D, ond C, Spitzer, Am. J. Obstet. GYnecol.l62:~52-4 (1990)). In astudy which compared the quantatlve yieid of cellular material obtained by 2 5 various methods, cervicovaginal lavage was found to be superior to scrape, swab or brush methocls for obtaining cervicovaginal cells (Goldberg, (~L et. al., Am. J. Obstet. GYnecol~ 16l: l 169-72 (1989); V~rmund, SH et. ai., Am. J. Obstet.
Gvnecol. 160: 304-8 (1989)). However, cervicovaginal cells obtained by lavage are in a liquid format and therefore present the same problems with handling, ; 1 ` ' 30 proaessing and contamination as scsmples obtained by mouthwashing techniques.
~: ' j;
WO 94/031 1 1 ~ 1 3 7 2 ~ 2 P~/US93/0738 '~=
For forensic purposes. DNA has been o~taine~ from epithelial cells, spun from urine (Gaspc~rinin, P. et. al., N. Enal. J. Med. 320: 80~ (1989)); and from root and surrounding sheath cells obtained from plucked hairs (Higuchi, R. et. al., Nature 332:543-6 (1988)). However, samples so obtained often do not contain S enough DNA for genetic analysis. Therelore hair and urine samples are not ' optimal for clinical ana~sis.
A simple, noninvasive technique for obtaining a non-liquid cell sample having an appropriate amount of genetic material for clinical analysis ~ould be 1 0 useful.
Summar,r o~ the InvenHon The invention relates to a simple, noninvasive process 'or obtaining a non-liquid cell sample from an animal for genetic analysis. The process involves inserting an appropriately srzed brush into an animal's oral cavily, contacting the brush with an oral membrane, agr~ating the brush against the membrane so that cells are captured within the bristles of the brush, removing the brush, which contains the captured ce!ls and isolating the cells from the bristles of the brush for use in genetic analysis. In a preferred embodiment, the brush is spiral, so thatcells can be obtained by rotating the brush against an oral mernbrane.
Cells so obtaineci can then be analyzed using any of a number of known genetic techniques. For example, cells obtained according to the disclosed method can be Iysed 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 arlalysis.
A major advantage of using the process described herein for obtaining ~ I 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 prane to contamination (e.g by bacteria~ and is more stable.
, J--`-'94~03ll~ 21~7X82 Pcr/US93/0738'`
l` ` '.
Detailed ~e~ ot the Invention In general, the invention relates to a process for obtainlng 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. Exarr ples of cells which can be obtained from an animal's oral cavity include epithelial cells of the inner cheek or tongue.
Accordin~ to the method of the invention, a brush is inserted into an anlmal'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.
To be suitable for use in the subject invention, a brush must be of an appropriate size to enable insertion into the animal's oral cavity. In addition, the brush bristles should be free of potentially contaminating material. For example, a brush comprising mammalian hair should not be used in the subject method, 2 0 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 docron) bristles. An especially preferr~d brush for use in ths invention is the nylon bristle cytology brush, (CytosottTM Medical Packaging Corporation, Camarillo, CA), which is currently 2 5 used for Pap Smear and Chlamydia testing.
Once removed from the mouth, a brush which contains captured oral cells can be analyzed directly or transported andlor stored for subsequent genetic analysis. At thls point, measures aimed at avoiding the dislodging of ~ I captured cells from a brush should be taken. For example, the brush can be pl~ced in a container (e.g. an envelope) during transportation and/or storage.
Cells may t~ obtained from a brush by any method which avoids loss of captured cells. A preferred method of obtaining cells from th t brush is by :
,:
,.~ , ~ ~ .
:`
WO 94/031 1 1 PCr/US93/0738'~
2137282 '~
contacting the cells with a solution. For example, if DNA is to be isolated from the cells for amplification, the cells c~n be obtained from the brush using a solution which Iyses cells (e.~. sodium hydroxide, potassium hydroxide).
Genetic material from oral celis so obtained can then be analyzed genetically using any of a number of known genetic techniques, such as di~estion with restriction endonucleases, ultraviolet light visualization of ethidium i~romld~ stalned a~arose g81s, DNA sequencing, or hybridization with detectable nucloic acid sequences ~e.~, labelled probes). In addi~ion, the genetic materialfrom cells obtalned accordins to the disclosed method can be amplified using known arnplification techniques, such as Q-beta-replicase and the polymerase chain reaction (PCR) (Saiki, R.K. et. al~, Science 239:487-491 (1~88); Wong, C., et.
al., Nature 330, 384-386 ~1987~ and U.S. Patent 4,683,202 to Mullis et. al.) to further increase the amount of genetic material available for genetic analysis.
The invention will now be illustrateci in the following Example, which is not intended to be limiting in any way.
Example 1: Obtaining Cells From Oral Cavities Using either a Brush or a Wooden 2 0 Spatula The oral cavities of five human subJects were scraped with wooden spatulas for about 30 seconds, and in five human subjects, the CytosottTM nylon bristle cytology brushes (Medical Packa~ing Corporation; Camarilo, CA) were 2 5 rotatad 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, a~ter whlch one tenth volume of 1 M TRIS (pH8) was added to the test tubes to neutralke.
3 0 , I The cheok c811 Iysates obtained using the spatulas or the brushes were used directiy in à po!ymerase chain reaction ~PCR) to amplify one or mofe of théfollowing exons of the Cystic Fibrosis transmembrane regulator gene: exons 4, 10, 11, 20 anci 21. Apropriats primers for the PCR were deYeloped based on the nuclelc acid sequence cilsclosed in Riordan, J.R. et al., Science 245:1066-1073 ~1989).
~`-` ') 94/03]i 1 1 PCI`/US93/07382 . l `` 2 t~282 PCR amplification reactions were performed using AmpliTaq DNA
polymerase and a DNA Thermal Cycler (Perkin Elmer). Cheek Iysates were jl amplified for 28 cy~les of 1û seconcis at 94, 10 seconds 5~C, and 30 seconds at 74C, with a final soak for 10 minutes at 74C. The reaction buffer used was described previousiy (Saiki, R.K. et ai., Science 239:487-491 {1988)~.
Ampiified materiai was then blotted onto nyion membranes (Biotrans Plus, iCN i3iomedical) and PCR products were analyzed on 1% agarose gels. No PCR
products were detected for any of the five samples in which cells were obtained usin~ a wooden spatula. In contrast, PCR products were detected for all five samples obtained using a brush. These PCR products were hybridized to sequence specific probes to identify known mutations (Kerem. B. et al., Science 245:1073-1080 (1989); Lemna et al., N. Ena. J. Med. 322:291-296 (1990); Kerern et ai., Proc. Natl. Acad. Sci. USA 87:8447-8451 (1990).
Equivalents:
Those skilled in the art will recognize or be ab!e to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of 2 0 the invention dascribed specificaiiy herein. Such equivalents are intended to be encompassed In the scope of the foilowing cic~ims.
t'.
~: .
, ~
S
; : q : - 5 -` :~
~ ~,
~`-` ') 94/03]i 1 1 PCI`/US93/07382 . l `` 2 t~282 PCR amplification reactions were performed using AmpliTaq DNA
polymerase and a DNA Thermal Cycler (Perkin Elmer). Cheek Iysates were jl amplified for 28 cy~les of 1û seconcis at 94, 10 seconds 5~C, and 30 seconds at 74C, with a final soak for 10 minutes at 74C. The reaction buffer used was described previousiy (Saiki, R.K. et ai., Science 239:487-491 {1988)~.
Ampiified materiai was then blotted onto nyion membranes (Biotrans Plus, iCN i3iomedical) and PCR products were analyzed on 1% agarose gels. No PCR
products were detected for any of the five samples in which cells were obtained usin~ a wooden spatula. In contrast, PCR products were detected for all five samples obtained using a brush. These PCR products were hybridized to sequence specific probes to identify known mutations (Kerem. B. et al., Science 245:1073-1080 (1989); Lemna et al., N. Ena. J. Med. 322:291-296 (1990); Kerern et ai., Proc. Natl. Acad. Sci. USA 87:8447-8451 (1990).
Equivalents:
Those skilled in the art will recognize or be ab!e to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of 2 0 the invention dascribed specificaiiy herein. Such equivalents are intended to be encompassed In the scope of the foilowing cic~ims.
t'.
~: .
, ~
S
; : q : - 5 -` :~
~ ~,
Claims (9)
1. A process for obtaining a non-liquid animal cell sample comprising the steps of:
a) inserting a brush having bristles and capable of being inserted into an animal's oral cavity;
b) contacting the bristles of the brush with an oral membrane comprising ceils:
c) agitating the brush against the membrane so that ceils from the membrane are captured in the brush bristles;
d) removing the brush containing captured cells from the oral cavity.
thereby obtaining a non-liquid cell sample.
a) inserting a brush having bristles and capable of being inserted into an animal's oral cavity;
b) contacting the bristles of the brush with an oral membrane comprising ceils:
c) agitating the brush against the membrane so that ceils from the membrane are captured in the brush bristles;
d) removing the brush containing captured cells from the oral cavity.
thereby obtaining a non-liquid cell sample.
2. A process of Claim 1 wherein the brush is spiral.
3. A process of Claim 1 wherein the animal is a human.
4. A non-liquid cell sample obtained according to the method of Claim 1.
5. A process for conducting a genetic analysis of an animal comprising the steps of:
a) inserting a brush having bristles and capable of being inserted into an animal's oral cavity;
b) contacting the bristles of the brush with an oral membrane comprising cells;
c) agitating the brush against the membrane so that cells from the membrane are captured in the brush bristles;
d) removing the brush containing captured cells from the oral cavity;
and e) lysing the cells obtained in step d), thereby yielding genetic material;
f) conducting a genetic analysis on the genetic material.
? 94/03111 PCT/US93/07382
a) inserting a brush having bristles and capable of being inserted into an animal's oral cavity;
b) contacting the bristles of the brush with an oral membrane comprising cells;
c) agitating the brush against the membrane so that cells from the membrane are captured in the brush bristles;
d) removing the brush containing captured cells from the oral cavity;
and e) lysing the cells obtained in step d), thereby yielding genetic material;
f) conducting a genetic analysis on the genetic material.
? 94/03111 PCT/US93/07382
6. A process of Claim 5 wherein the animal is a human.
7. Genetic material obtained according to the method of Claim 5.
8. A process for detecting the presence of a particular nucleic acid sequence of interest in genetic material contained in cells obtained from an animal, comprising the steps of:
a) inserting a brush having bristles and capable of being inserted into a mammal's oral cavity;
b) contacting the bristles of the brush with an oral membrane comprising cells;
c) agitating the brush against the membrane so that cells from the membrane are captured in the brush bristles;
d) removing the brush containing captured cells from the oral cavity;
e) lysing the cells obtained in step d), thereby yielding genetic material;
g) contacting the genetic material with a detectable nucleic acid sequence complementary to the nucleic acid sequence of interest;
h) allowing the genetic sequences and the detectable nucleic acid sequence to hybridize; and i) detecting hybridization as an indication of the presence of the nucleic acid sequence of interest.
a) inserting a brush having bristles and capable of being inserted into a mammal's oral cavity;
b) contacting the bristles of the brush with an oral membrane comprising cells;
c) agitating the brush against the membrane so that cells from the membrane are captured in the brush bristles;
d) removing the brush containing captured cells from the oral cavity;
e) lysing the cells obtained in step d), thereby yielding genetic material;
g) contacting the genetic material with a detectable nucleic acid sequence complementary to the nucleic acid sequence of interest;
h) allowing the genetic sequences and the detectable nucleic acid sequence to hybridize; and i) detecting hybridization as an indication of the presence of the nucleic acid sequence of interest.
9. A process of Claim 8 wherein the animal is a human.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US92987192A | 1992-08-07 | 1992-08-07 | |
| US07/929,871 | 1992-08-07 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2137282A1 true CA2137282A1 (en) | 1994-02-17 |
Family
ID=25458604
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002137282A Abandoned CA2137282A1 (en) | 1992-08-07 | 1993-08-05 | Process for obtaining a non-liquid cell sample |
Country Status (4)
| Country | Link |
|---|---|
| EP (1) | EP0653920A1 (en) |
| JP (1) | JPH08501228A (en) |
| CA (1) | CA2137282A1 (en) |
| WO (1) | WO1994003111A1 (en) |
Families Citing this family (6)
| 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 (en) * | 1995-11-15 | 2000-07-05 | Genelink Inc | Non-invasive identification system |
| JP4748628B2 (en) * | 1999-02-23 | 2011-08-17 | シーディーエックス ラボラトリーズ インコーポレーテッド | A minimally invasive device for examining oral and similar epithelial lesions |
| EP3669747B1 (en) | 2014-05-02 | 2022-03-30 | United States Endoscopy Group, Inc. | Cleaning device for an endoscopic device |
| JP2016045103A (en) * | 2014-08-25 | 2016-04-04 | 日東電工株式会社 | Sample sampling material for gas chromatography-mass spectrometry |
| JP2016045102A (en) * | 2014-08-25 | 2016-04-04 | 日東電工株式会社 | Sample sampling material for high-performance liquid chromatography |
Family Cites Families (3)
| 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 |
| AU643086B2 (en) * | 1990-10-26 | 1993-11-04 | Sangtec Molecular Diagnostics Ab | Method for evaluating the adequacy of clinical specimens for hybridization assays and kit for performing the evaluation |
-
1993
- 1993-08-05 EP EP93919898A patent/EP0653920A1/en not_active Withdrawn
- 1993-08-05 JP JP6505572A patent/JPH08501228A/en active Pending
- 1993-08-05 CA CA002137282A patent/CA2137282A1/en not_active Abandoned
- 1993-08-05 WO PCT/US1993/007382 patent/WO1994003111A1/en not_active Application Discontinuation
Also Published As
| Publication number | Publication date |
|---|---|
| EP0653920A1 (en) | 1995-05-24 |
| WO1994003111A1 (en) | 1994-02-17 |
| JPH08501228A (en) | 1996-02-13 |
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