WO1999057310A2 - Instrument d'analyse et de diagnostic - Google Patents
Instrument d'analyse et de diagnostic Download PDFInfo
- Publication number
- WO1999057310A2 WO1999057310A2 PCT/EP1999/002918 EP9902918W WO9957310A2 WO 1999057310 A2 WO1999057310 A2 WO 1999057310A2 EP 9902918 W EP9902918 W EP 9902918W WO 9957310 A2 WO9957310 A2 WO 9957310A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- analysis chip
- analysis
- matrix
- chip
- biomolecule
- Prior art date
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
Definitions
- the invention relates to an analysis and diagnostic instrument for examining biological samples, in particular DNA molecules.
- analysis chips A number of analytical and diagnostic instruments are known in the field of biological and medical laboratory technology. In certain configurations, these are usually also referred to as analysis chips. If the analysis chip is intended for the investigation of DNA molecules, it is called a DNA analysis chip, DNA array or DNA grid.
- analysis chip and analysis and diagnostic instrument are used synonymously in the following.
- Conventional analysis chips usually consist essentially of a carrier material, in the surface of which wells are etched, in which, in the case of a DNA analysis, various DNA molecules are incorporated.
- hybridization By bringing a DNA sample to be examined into contact with the bound DNA molecules can lead to hybridization, which provide signals about the presence or absence and possibly the concentration of nucleic acids in the hybridization solution.
- a special analyzer then reads out the hybridization data.
- the detection of the hybridization reaction is conventionally carried out by optical methods, such as by coupling light through a grating. The mass accumulation disturbs the evanescent field, whereby a disturbance in the far field can be measured.
- Conventional analysis chips generally use silicon as the carrier material, which contains a non-transparent medium on which the DNA molecules are bound in the wells.
- Analog, not highly integrated systems, such as ELISA systems have in particular the disadvantages that the biological sample to be examined has to be applied individually to each reference molecule in the wells, the coupling of light to detect the hybridization reaction is inefficient, and an adjustment of the analysis chip in the analyzer is difficult , the administration and processing of data and parameters is complex and the costs for production and operation are very high.
- the object of the invention is therefore to provide an improved analysis or diagnostic instrument or method.
- the invention is based on the basic idea of providing an analysis chip for examining biological samples, which essentially consists of a carrier, on the surface of which at least one biomolecule matrix composed of individual dots on a bottom of a microfluidic structure, such as, for example a meander structure, or a planar surface is applied.
- the biological sample to be examined is applied to one end of the microfluidic structure and flows along this structure to the other end, the sample flowing over the respective matrix points, each of which has an individual represent viduelle molecular species, triggers certain reactions.
- this reaction is usually a hybridization reaction.
- molecules can be detected in the sample which have a greater or lesser degree of similarity or homology with the complementary molecular species on the surface or a part thereof.
- hybridization conditions depend, for example, on the temperature or the salt concentration in the sample and can be set by the person skilled in the art using customary methods. For example, the salt concentration in a sample can be adjusted.
- the setting of hybridization conditions is known in the prior art and can e.g. based on the teaching in Harnes and Higgins, "Nucleic acid hybridization, a practical approach", IRL Press, Oxford 1985.
- the molecule to be detected in the biological sample is a protein or peptide
- a number of possibilities are available to the person skilled in the art for how he can detect this with the analysis chip according to the invention.
- One possibility is the incorporation of the molecule into an antibody sandwich, the antibody (or a derivative thereof) that is not fixed to the surface preferably being marked in a detectable manner.
- biological sample means in the broadest sense that the sample contains biological material such as nucleic acids or proteins or derivatives or fragments thereof.
- nucleic acids can be modified by methods of recombinant DNA technology. adorned, fragmented, etc.
- proteins may have been enriched from a natural source prior to analysis.
- the sample is transferred from a natural source directly to the analysis.
- the analysis chip also has a storage medium for easier data management and processing.
- the analysis or diagnostic instrument has the advantages over the prior art in particular that the sample to be examined only has to be applied to the microfluidic structure at one point, as a result of which the process time can be considerably reduced and it can be produced in a simple and inexpensive manner , since the microfluidic structure can be produced simultaneously with the carrier, the theoretical signal strength can be increased by the microstructuring in the area of the samples, a more efficient reaction detection is possible, and / or production-related and / or other data, such as Configuration, production lot, zero control data, patient data, expiry date, analysis results and the like can be saved directly on the analysis chip.
- Another advantage is that the results of the analysis can be determined with considerably greater certainty due to a matrix that is preferably at least double redundant.
- the analysis is carried out with a high integration density and high parallelism. Only small volumes of the samples are required. Further advantages are the miniaturization of the analysis chip, the possibility of real-time and / or online evaluation at the same time as the hybridization reaction.
- FIG. 1 shows a spatial image of an analysis chip according to the invention
- FIG. 2 shows a section along the surface II -II of FIG. 1 to illustrate the coupling of light for the detection of fluorochrome molecules
- 3 shows the detail III from FIG. II;
- Fig. 4 shows the principle of light coupling for the detection of mass accumulation.
- the analysis or diagnostic instrument 2 - in short analysis chip - shown in Fig. 1 consists essentially of a carrier or support frame 4, which in cross section e.g. has a U-shaped profile which is closed at its ends, so that a cavity 8 is provided under a surface 6.
- a microfluidic structure e.g. a meander structure 10 is provided, which is embedded in the carrier 6 or preferably in at least one preferably transparent medium 12 and 14.
- This transparent medium 12 and 14 is e.g. an insert made of glass, quartz glass, plastic, silicon, silicates.
- the meander structure 10 is designed such that dots 16 of a biomolecule matrix can be applied therein, each point 16 of the matrix preferably representing an individual molecular species (DNA, RNA, proteins, peptides, polysaccharides, etc.) At least one of the dots 16 is present on the analysis chip 2, namely on the medium 12, but preferably at least twice redundantly, ie also at least on the medium 14 or other media (not shown).
- the surface of the analysis chip or the medium 12 and 14 can be planar (without a microfluidic structure) and to provide the at least one biomolecule matrix thereon.
- a surface opposite the biomolecule matrix which contains the fluid volume with the sample to be examined, has the microfluidic structure.
- the analysis chip 2 then becomes Analysis placed on this surface.
- the microfluidic structure 10 can furthermore be provided both on the surface 6 of the analysis chip 2 and on the opposite surface.
- the carrier 4 of the analysis chip is preferably a plastic, which is produced by micro injection molding.
- Derivatives of polycarbonate are particularly suitable as carrier materials.
- the covalent coupling of biomolecules is preferably carried out on semifluidic, gel-like polymer surfaces in the area of the samples (e.g. polymer brush).
- the polymer surfaces have their own hydrodynamics, which gives the polymer surfaces special hydrodynamic properties and has a positive influence on the relationship between signal and background noise.
- Semifluidic polymer surfaces are synthesized directly on the surface 6 of the analysis chip and can generally be used as nano- to microstructuring of surfaces. are referred to as polymer brushes.
- the theoretical signal strength is enhanced by the semi-fluidic surface structuring in the area of the samples.
- genetic material is isolated from a biological sample to be examined, specifically amplified and, if necessary, marked.
- the probe material obtained in this way is applied to one end of the meander structure 10 of the analysis chip 2 by means of a device, for example a pipette or syringe.
- the biological sample to be examined or the genetic material flows over the individual points 16 of the biomolecule matrix along the meandering structure 10, each of which represents an individual molecule species, and hybridizes in the case of at least partially complementary biomolecules.
- a number of methods are available to the person skilled in the art for analyzing and evaluating the data of the analysis chip 2. Two basic methods are preferred according to the invention.
- the evaluation can be carried out by a real-time measurement of the hybridization by detecting the mass accumulation by providing the analysis chip with a grating coupler, which causes a disturbance in the evanescent field.
- the evaluation can be carried out by a real-time measurement of the hybridization by detecting the mass accumulation by providing the analysis chip with a grating coupler, which causes a disturbance in the evanescent field.
- the analysis chip according to the invention shown in FIGS. 1 to 3 is particularly suitable for fluorochrome-marked systems.
- the analysis chip shown in FIG. 4 is suitable for evaluation with a hybridization detection by mass accumulation.
- the analysis chip 2 has adjustment means 18 on the outer edge of the carrier 4, e.g. Adjusting lugs or indentations, so that when inserted into an analyzer (not shown) for evaluating the examination is precisely adjustable.
- adjustment means 18 on the outer edge of the carrier 4, e.g. Adjusting lugs or indentations, so that when inserted into an analyzer (not shown) for evaluating the examination is precisely adjustable.
- the individual dots 16 of the biomolecule matrix as well as the loops of the meandering structure 10 are preferably themselves relatively close together, e.g. with a distance between 100 and 500 ⁇ m. It is ensured that the individual points 16 of the analysis chip 2 correspond to corresponding evaluation elements of the analyzer.
- the analysis chip 2 also has at least one storage medium which can store operating data, production-relevant data such as configuration, production lot, zero control data, entered patient data, the expiry date, analysis results and other auxiliary data.
- the storage medium is preferably an electronic one Memory chip 20, a magnetic strip 22 and / or a bar code 24, however, can also be any other known storage medium, such as ROM or RAM.
- the analysis chip 2 shown in FIGS. 2 and 3 has, in addition to the features described with reference to FIG. 1, the features which are advantageous for the detection of fluorochrome molecules.
- the analysis of a biological sample in the meander structure 10 of the analysis chip 2 takes place in that in an analyzer the analysis chip 2 from the side facing away from the meander structure 16, i.e. is illuminated by the cavity 8, which is shown schematically in the form of arrow 26.
- the underside 28 of the surface 6 is preferably provided with an optical lens field 30, which enables confocal illumination.
- the lens array 30 is preferably designed such that it is located exactly below the meandering structure 10, in particular exactly below the dots 16, so that the illumination of the individual dots 16 shown in FIG. 3 is made possible.
- the lens field 30 and the carrier 4 of the analysis chip 2 can consist of a uniform material, which in one step e.g. can be produced by micro injection molding. However, the lens field 30 is preferably located on the underside 28 of the transparent medium 12 or 14 inserted into the carrier 4.
- the illumination 26 from the underside 28 of the analysis chip 2 has the particular advantage that the intensity of the light source can be reduced since the light beams do not have to pass through a beam splitter.
- the incident light 26 is focused by the lenses of the lens array 30 onto the individual points 16 of the biomolecule matrix, as a result of which the dots 16, which have caused a reaction with the sample to be examined, cause fluorescence through the fluorochrome molecules.
- Other ways to prove the Molecules are based on the principle of bioluminescence, eg phosphorescence, radioactive labels, which can be detected by applying X-ray films.
- a suitable evaluation circuit for example an optoelectronic circuit, can then be used to further process the data obtained with respect to the sample to be analyzed. This data can be further processed together with the data stored in the storage medium and / or written into the memory of the analysis chip 2.
- FIG. 4 shows the coupling of light for analysis chips in which the hybridization detection takes place via the detection of the mass accumulation.
- an optical method is used in which the light is coupled in via a large number of gratings 32, as a result of which the evanescent field is influenced. This influence can be measured in the far field 39.
- Exactly one optical grating 32 is assigned to each point 16 of the biomolecule matrix in the microfluidic structure 10, so that the efficiency of the light coupling can be increased considerably.
- the light 34 from an illumination source is focused on the grating 32 arranged in the matrix via a two-dimensional lens field 36 or via a two-dimensional Fresnel lens field, for example a hologram or diffractive optics.
- the analysis data obtained can be further processed and / or written into the memory of the analysis chip 2 together with the data stored in the storage medium in an analyzer (not shown) by using suitable evaluation circuits, for example optoelectronic circuits.
- the optical gratings 32 were preferably introduced into the microfluidic structure 10 together with the respective points 16 of the biomolecule matrix, which each represent an individual molecular species (DNA, RNA, proteins, peptides, polysaccharides, etc.).
- the biological sample to be examined passes through the microfluidic structure 16 (meandering structure), for example in the direction of the arrows 38.
- the lens field 36 is located on the side of the grid 32 facing away from the biomolecule matrix. It can, similarly as previously with reference to FIGS. 2 and 3 described, also be provided in the cavity 8 of the analysis chip 2. Likewise, it can consist of a uniform material together with the carrier 4, or, preferably, be provided on the transparent media 12 and 14.
- analysis chip according to the invention is also suitable for further evaluation methods not described in detail.
- microfluidic structure 16 as a meandering structure is only exemplary; rather, any other microfluidic structures, such as e.g. a tree structure possible.
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- Health & Medical Sciences (AREA)
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
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- Biomedical Technology (AREA)
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- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
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- Analytical Chemistry (AREA)
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Abstract
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AU38252/99A AU3825299A (en) | 1998-04-30 | 1999-04-29 | Instrument for analysis and diagnostics |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE19819537.0 | 1998-04-30 | ||
DE1998119537 DE19819537A1 (de) | 1998-04-30 | 1998-04-30 | Analyse- und Diagnostikinstrument |
Publications (2)
Publication Number | Publication Date |
---|---|
WO1999057310A2 true WO1999057310A2 (fr) | 1999-11-11 |
WO1999057310A3 WO1999057310A3 (fr) | 2000-03-09 |
Family
ID=7866413
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/EP1999/002918 WO1999057310A2 (fr) | 1998-04-30 | 1999-04-29 | Instrument d'analyse et de diagnostic |
Country Status (3)
Country | Link |
---|---|
AU (1) | AU3825299A (fr) |
DE (1) | DE19819537A1 (fr) |
WO (1) | WO1999057310A2 (fr) |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2003008974A1 (fr) * | 2001-07-18 | 2003-01-30 | Micronas Gmbh | Biodetecteur et procede de detection d'analytes grace a la luminescence a resolution temporelle |
WO2003080789A1 (fr) * | 2002-03-27 | 2003-10-02 | Micronas Gmbh | Dispositif et procede de detection de processus cellulaires par des mesures de luminescence |
WO2003082730A1 (fr) * | 2002-03-31 | 2003-10-09 | Gyros Ab | Dispositifs microfluidiques efficaces |
WO2003104384A1 (fr) * | 2002-06-05 | 2003-12-18 | Bioprocessors Corporation | Systemes reacteurs pourvus d'un composant interagissant avec la lumiere |
WO2004052540A3 (fr) * | 2002-12-05 | 2004-09-16 | Protasis Corp | Ensemble de substrat micro-fluidique adaptable |
WO2007050040A1 (fr) * | 2005-10-28 | 2007-05-03 | Agency For Science, Technology And Research | Unité d'immobilisation et dispositif pour l'isolement de molécules d'acides nucléiques |
Families Citing this family (9)
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JP2001186881A (ja) * | 1999-10-22 | 2001-07-10 | Ngk Insulators Ltd | Dnaチップの製造方法 |
DE10120035B4 (de) | 2001-04-24 | 2005-07-07 | Advalytix Ag | Verfahren und Vorrichtung zur Manipulation kleiner Flüssigkeitsmengen auf Oberflächen |
DE10312670A1 (de) * | 2003-03-21 | 2004-10-07 | Friz Biochem Gmbh | Substrat zur kontrollierten Benetzung vorbestimmter Benetzungsstellen mit kleinen Flüssigkeitsvolumina, Substratabdeckung und Flusskammer |
US8221843B2 (en) | 2005-02-15 | 2012-07-17 | Lisa Dräxlmaier GmbH | Methods and compositions for coating interior components of motor vehicles and interior components of motor vehicles coated using same |
DE102005010024B4 (de) | 2005-03-04 | 2015-06-11 | Lisa Dräxlmaier GmbH | Fahrzeugtürverkleidung und Verfahren zu deren Herstellung |
DE102005010025B4 (de) | 2005-03-04 | 2008-01-24 | Lisa Dräxlmaier GmbH | Fahrzeugtürverkleidung und Verfahren zu deren Herstellung |
DE102005013477B4 (de) | 2005-03-23 | 2008-12-04 | Lisa Dräxlmaier GmbH | Kraftfahrzeugverkleidungsteil mit Schwächungsstellen und Verfahren zu dessen Herstellung |
DE102005034354B4 (de) | 2005-07-22 | 2014-08-28 | Lisa Dräxlmaier GmbH | Airbagabdeckung |
DE102021212505A1 (de) | 2021-11-08 | 2023-05-11 | Robert Bosch Gesellschaft mit beschränkter Haftung | Optische Vorrichtung zum Anregen einer Probe, Analysegerät und Verfahren zum Anregen einer Probe |
Citations (8)
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EP0535242A1 (fr) * | 1991-03-18 | 1993-04-07 | Institut Molekulyarnoi Biologii Imeni V.A. Engelgardta Akademii Nauk Sssr | Procede et dispositif de determination de la sequence de nucleotides d'adn |
US5284622A (en) * | 1991-10-02 | 1994-02-08 | Boehringer Mannheim Gmbh | Test carrier for the analysis of fluids |
US5661028A (en) * | 1995-09-29 | 1997-08-26 | Lockheed Martin Energy Systems, Inc. | Large scale DNA microsequencing device |
US5770721A (en) * | 1993-08-11 | 1998-06-23 | University Of Chicago | Method of manufacturing a matrix for the detection of mismatches |
WO1998045481A1 (fr) * | 1997-04-04 | 1998-10-15 | Caliper Technologies Corporation | Analyseurs biochimiques fonctionnant en boucle fermee |
WO1998056956A1 (fr) * | 1997-06-09 | 1998-12-17 | Caliper Technologies Corporation | Dispositifs et procedes pour la correction des variations de vitesse dans les systemes microfluidiques |
US5851772A (en) * | 1996-01-29 | 1998-12-22 | University Of Chicago | Microchip method for the enrichment of specific DNA sequences |
WO1999015888A1 (fr) * | 1997-09-19 | 1999-04-01 | Aclara Biosciences, Inc. | Appareil et procede d'electrofluidite capillaire |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE3709296C1 (de) * | 1987-03-20 | 1988-09-22 | Draegerwerk Ag | Kolorimetrisches Gasmessgeraet |
DE19621179C2 (de) * | 1996-05-25 | 2000-09-07 | Nonnenmacher Klaus | Transporteinheit für Proben |
JP3739537B2 (ja) * | 1997-03-26 | 2006-01-25 | 大日本印刷株式会社 | 光学的分析装置用測定チップ |
-
1998
- 1998-04-30 DE DE1998119537 patent/DE19819537A1/de not_active Ceased
-
1999
- 1999-04-29 WO PCT/EP1999/002918 patent/WO1999057310A2/fr active Application Filing
- 1999-04-29 AU AU38252/99A patent/AU3825299A/en not_active Abandoned
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0535242A1 (fr) * | 1991-03-18 | 1993-04-07 | Institut Molekulyarnoi Biologii Imeni V.A. Engelgardta Akademii Nauk Sssr | Procede et dispositif de determination de la sequence de nucleotides d'adn |
US5284622A (en) * | 1991-10-02 | 1994-02-08 | Boehringer Mannheim Gmbh | Test carrier for the analysis of fluids |
US5770721A (en) * | 1993-08-11 | 1998-06-23 | University Of Chicago | Method of manufacturing a matrix for the detection of mismatches |
US5661028A (en) * | 1995-09-29 | 1997-08-26 | Lockheed Martin Energy Systems, Inc. | Large scale DNA microsequencing device |
US5851772A (en) * | 1996-01-29 | 1998-12-22 | University Of Chicago | Microchip method for the enrichment of specific DNA sequences |
WO1998045481A1 (fr) * | 1997-04-04 | 1998-10-15 | Caliper Technologies Corporation | Analyseurs biochimiques fonctionnant en boucle fermee |
WO1998056956A1 (fr) * | 1997-06-09 | 1998-12-17 | Caliper Technologies Corporation | Dispositifs et procedes pour la correction des variations de vitesse dans les systemes microfluidiques |
WO1999015888A1 (fr) * | 1997-09-19 | 1999-04-01 | Aclara Biosciences, Inc. | Appareil et procede d'electrofluidite capillaire |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2003008974A1 (fr) * | 2001-07-18 | 2003-01-30 | Micronas Gmbh | Biodetecteur et procede de detection d'analytes grace a la luminescence a resolution temporelle |
WO2003080789A1 (fr) * | 2002-03-27 | 2003-10-02 | Micronas Gmbh | Dispositif et procede de detection de processus cellulaires par des mesures de luminescence |
US7767443B2 (en) | 2002-03-27 | 2010-08-03 | Micronas Gmbh | Device and method for detecting cellular processes by means of luminescence measurements |
WO2003082730A1 (fr) * | 2002-03-31 | 2003-10-09 | Gyros Ab | Dispositifs microfluidiques efficaces |
WO2003104384A1 (fr) * | 2002-06-05 | 2003-12-18 | Bioprocessors Corporation | Systemes reacteurs pourvus d'un composant interagissant avec la lumiere |
WO2004052540A3 (fr) * | 2002-12-05 | 2004-09-16 | Protasis Corp | Ensemble de substrat micro-fluidique adaptable |
WO2007050040A1 (fr) * | 2005-10-28 | 2007-05-03 | Agency For Science, Technology And Research | Unité d'immobilisation et dispositif pour l'isolement de molécules d'acides nucléiques |
Also Published As
Publication number | Publication date |
---|---|
AU3825299A (en) | 1999-11-23 |
DE19819537A1 (de) | 2000-03-16 |
WO1999057310A3 (fr) | 2000-03-09 |
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