EP2596129A1 - Verfahren, vorrichtung und testkit für molekularbiologische reaktionen - Google Patents
Verfahren, vorrichtung und testkit für molekularbiologische reaktionenInfo
- Publication number
- EP2596129A1 EP2596129A1 EP11751837.3A EP11751837A EP2596129A1 EP 2596129 A1 EP2596129 A1 EP 2596129A1 EP 11751837 A EP11751837 A EP 11751837A EP 2596129 A1 EP2596129 A1 EP 2596129A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- reaction
- components
- pcr
- amplification
- carrying
- 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- Withdrawn
Links
Classifications
-
- 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
- C12Q1/6804—Nucleic acid analysis using immunogens
-
- 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
- C12Q1/6813—Hybridisation assays
- C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
-
- 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6848—Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
-
- 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
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
-
- 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
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
-
- 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
- C12Q2565/00—Nucleic acid analysis characterised by mode or means of detection
- C12Q2565/60—Detection means characterised by use of a special device
- C12Q2565/625—Detection means characterised by use of a special device being a nucleic acid test strip device, e.g. dipsticks, strips, tapes, CD plates
Definitions
- the invention relates to reagent components for carrying out molecular genetic investigations, in particular under field conditions. These reagent formulations are also storage stable at ambient temperature. Possible applications include polymerase chain reaction (PCR), reverse transcription (RT) enzyme-substrate, antigen-antibody interactions, and protein synthesis.
- PCR polymerase chain reaction
- RT reverse transcription
- a number of different components is required. These are e.g. Amplification buffers, salts, enzymes, dNTPs, nucleic acids, etc.
- all components for the detection reaction including the biomolecules, must be stable over a longer period of time, if possible.
- a compilation of multiple solutions (kit components) will only last as long as its most delicate component.
- This critical component is often biocatalysts or proteins that must be present in native and / or active form according to the application.
- dNTPs nucleotides
- polydextrose, mannitol or sorbitol may also be used as humectants be used because they functionally replace water.
- This type of stabilization is also often linked to a cooling possibility, so that the degradation of the biomolecule over a longer period can not be excluded.
- Successful storage of enzyme preferably a Taq HOT start polymerase
- primer, probe, dNTPs and an internal positive control is described in US patent application US 02005 006 98 98A1.
- the corresponding chemicals were applied together with HEPES on a bead with mannitol, which is dissolved in water for PCR and then used.
- 4,891,319 describes the protection of proteins and other biological molecules by adding trehalose to an aqueous solution, thus achieving successful stabilization while maintaining the activity of the components, for example, for 6 months at 40 ° C for blood coagulation factors (US Patent 7501493). and other molecules (Paz-Alfaro KJ, Ruiz-Granados YG, Uribe-Carvajal S, Sampedro JG, Trehalose-mediated thermal stabilization of glucose oxidase from Aspergillus niger. J Biotechnol., 2009 May 20; 141 (3-4): 130 Epub 2009 Mar 25 and Morana A, Stiuso P, Colonna G, Lamberti M, Carteni M, De Rosa M.
- biomolecules can be freeze-dried and rehydrated before use. However, here too the dried biomolecules must be stored cool. In this connection are also the 2.6 nm large, lyophilized particles of the company Cepheid (Sunnyvale, CA) mentions.
- dNTPs, polymerase, magnesium chloride and buffer are combined in one particle and can be used for further applications (PCR) after addition of water and primers.
- Cepheid relies on a freeze-drying process developed by ABAXIS Inc., the so-called "Orbos Technology" (http://www.abaxis.com/about_us/history.html) .Disadvantage of the method of drying is the rather high equipment costs for a freeze-drying.
- the Ready-to-go product line is a partially stabilized PCR component, with small beads containing Taq polymerase, dNTPs, and amplification buffer, which are stable at room temperature for an extended period of time is that the primer and probe must be freshly added by the customer, which makes this method unsuitable for use outside of a laboratory
- Klatser et al Klatser et al (Klatser PR, Sjoukje Kuijper, van Ingen CW, Kolk AHL Stabilized, Freeze-dried PCR Mix for detection of mycobacteria. J Clin Microb, 1998 Jun 36 (6): 1798-1800 1998).
- primers, buffers, dNTPs, uracil DNA glycosylase and polymerase are freeze-dried with 5% trehalose.
- trehalose When stored at 4 or 20 ° C, they were still active for up to one year after drying. Storage at 37 ° C reduced the storage time to 3 months or 56 ° C to 1 week. The authors used 2 different polymerases. It turned out that a higher content of glycerol negatively influences the storage stability since glycerol has a hygroscopic effect.
- the disadvantage here, however, is that trehalose inhibits a series of amplification reactions and in particular reactions which couple an amplification reaction with a probe hybridization. Thus, such an agent is in any way universally applicable.
- the stabilization of the reagents mediated by trehalose can be increased even more by adding prionex, a polypeptide from porcine skin collagen, as in this case.
- the stability of the reagents was four times as long after addition of the polypeptide as with trehalose alone, instead of 30 days at 37 ° C.
- Another method for stabilizing biomolecules is to bind them to a mold carrier. Via a coupling reaction with an amino group, the biomolecule stably and permanently binds to the carrier material and can thus be used for detection reactions.
- This method is patented and disclosed (European Patent Application EP0511559).
- a modified, also patented method for stable immobilization of antibodies U.S. Patent 4,948,836) and oligonucleotides (German Patent DE 10053553C2) is also described.
- the drying takes place partially after incubation in a protein-sugar solution. This process is very cumbersome and time consuming and can not be realized in this form for a mixture of different substances, as they are necessary for a PCR.
- dNTPs nucleotide triphosphates
- primers primers
- buffer an enzyme
- Taq or reverse transcriptase an enzyme
- dNTPs One of the critical components for the storage stability of the individual solutions are the dNTPs, which decompose at a physiological pH (pH 7.0 to 7.5) to corresponding diphosphates and monophosphates.
- pH 7.0 to 7.5 physiological pH
- the content of functional dNTPs already decreases by 2-3% after 10 days at 37 ° C.
- the dNTPs can be stored in the presence of an appropriate stabilizer.
- adenosine triphosphate could be confirmed as stabilizers in the presence of EDTA (pH 8.3 to 9.2), guanidine / aminoguanidine (pH 9.0 to 10.0) or glycerol / hydrogen phosphate (pH 3.7) (EP0941370 bl).
- EDTA pH 8.3 to 9.2
- guanidine / aminoguanidine pH 9.0 to 10.0
- glycerol / hydrogen phosphate pH 3.7
- the prior art also includes the documents US 3645853 A, DE 2800437 AI, DE1220083 A, WO 98/37229 AI, EP0733714A2, EP0823972 AI and EP0925113 AI.
- U.S. Patent 3,645,853 describes a diagnostic composition and method for detecting nitrate reduction.
- a spongy material containing at least four impregnated zones is disclosed.
- the compartmentalization is carried out by applying solutions individually to paper and then assembling them manually after complete drying. There is no compatibility by nature. Only from one side substance is replaced. Other components are not dissolved, but a direct color change occurs after mixing the solutions on the strip.
- DE 2800437 Al mentions a device for obtaining a non-toxic atmosphere for use in growing anaerobic micro-organisms, wherein the surface area and the permeability of a membrane are selected such that the rate of access of the liquid and thus the rate of generation of the gas atmosphere is controlled.
- EP 0 733 714 A2 describes a nucleic acid amplification method and apparatus.
- a sample is pumped from the sample application to the reaction area.
- the reaction region is preferably described as a continuous channel: "Although it is preferred to provide the sample area and reaction area in the form of a continuous, undivided channel as shown, it is within the scope of the present invention to provide partitions or barriers It is therefore within the scope of the invention to provide partitions or barriers between the decontamination zone and the amplification zone of the reaction area. " However, there is no intention for the authors to separate the products.
- the subject of EP0733714A2 is a diagnostic device in which the release and capture medium form a biphasic chromatographic substrate. Both media are separate and made of different materials.
- the capture medium has no other immobilized components and is used for adsorption.
- the release medium is a hydrophilic material for holding, releasing and transporting the immunological material. The application is carried out by capillary action of the test solutions. Mixing is not specifically prevented.
- EP 0 925 113 AI reports an apparatus and a method for storing and distributing biochemical reagents.
- Each reagent is arranged in a single room, with the rooms separated from each other.
- the reagent cartridge is a single item. Reagent cartridges are combined to cassette cartridge.
- the invention had the object to eliminate the disadvantages mentioned in the prior art.
- the present invention has provided a molecular diagnostic detection method which contains all the required critical reagent components in a form which:
- the spatial separation of the components involved in the amplification or reverse transcription has not been considered in any of the previously disclosed solution.
- the agent according to the invention precisely takes into account the spatial separation of the components which are required for the reactions and thus makes it possible to protect the components involved from one another. This approach thus makes it impossible to initiate non-specific reactions (e.g., formation of primer domains, nonspecific amplification under suboptimal conditions, etc.).
- primer or primer and probes are placed on a porous filter disc (eg made of polyethylene).
- a porous filter disc eg made of polyethylene.
- the components are dried (eg incubation at 37 ° C. for 1 h) or else lyophilized. Subsequently, the filter disc is stored dry. The release of the spent reaction components takes place in that the filter disk is supplied with an aqueous solution which contains all further reaction components required for the amplification reaction / detection reaction in the respectively optimum concentration (dNTPs, amplification buffer, magnesium and optionally further additives).
- This aqueous solution can be prepared in advance as a master mix and may already contain the nucleic acid to be examined.
- a detergent eg, an octylphenol ethoxylate or polysorbate
- This detergent improves the release of the components located on the filter disk.
- the required amount of dNTPs can additionally be compartmentalized on the filter disc and thus spatially separated from the other reaction components.
- the preparation of the storage-stable filter disc and the storage is carried out as described.
- the release of the spent reaction components again takes place by supplying to the filter disk an aqueous solution which contains all further reaction components required for the amplification reaction / detection reaction in the respectively optimum concentration.
- This aqueous solution can be prepared in advance and may already contain the nucleic acid to be examined.
- An advantage of this embodiment is that the dNTPs were also stored stable on the filter disk. Thus, this component of the aqueous solution to release the reaction components no longer needs to be included.
- the agent according to the invention can also be used for carrying out a reverse transcriptase reaction for the investigation of a target RNA.
- the reverse transcriptase, dNTPs, the concentrated buffer and primers required for the reaction are spent on the filter disk.
- RNA to be rewritten possibly a detergent and RNA-stabilizing reagents.
- this method has another advantage: The amount of RNA used can be considerably increased because the necessary components are not added as an aqueous solution but are dissolved directly in the RNA to be rewritten. This increases the sensitivity of the test method.
- the reaction buffer, the enzyme mix, the dNTPs and the primer and probe required for the reaction are spent and stabilized on the filter disc.
- the detachment of the components is carried out as in the case of a two-step PCR (cDNA synthesis and PCR in different reaction vessels) with an aqueous solution containing the RNA to be examined.
- compartmentalization and storage-stable immobilization of individual reaction components in different mixtures and enzyme-catalyzed reactions can be used. It is always ideal if the required reaction partners are not allowed to interact with each other before a reaction.
- reaction components which are required for a reaction in spatially separated arrangement can also be done without porous material on spatially separated areas of a vessel.
- the individual components are brought together by washing out with an aqueous solution immediately before the beginning of the reaction.
- the filter disc of the present invention provides greater protection against exogenous factors (e.g., oxygen, moisture, light).
- reaction molecular biological reactions
- biological molecules include any substances and compounds of essentially biological origin which possess properties which are scientifically, diagnostic and / or pharmaceutical applications are relevant.
- native molecules are included, as can be isolated from natural sources, but also derived forms, fragments and derivatives, as well as recombinant forms and artificial molecules, provided they have at least one property of the native molecules.
- RNA in DNA include the amplification, detection or transcription of nucleic acids, enzyme-substrate, antigen-antibody interactions and protein synthesis.
- PCR polymerase chain reaction
- RT reverse transcription
- primer or “probes” the skilled person understands oligonucleotides, wherein primers for amplification or reverse transcription are necessary. The singles can be marked. Labels of probes are known to those skilled in the art.
- enzymes includes polymerase and reverse transcriptase and other enzymes useful in molecular biology, such as peroxidases, dehydrogenases, phosphatases, etc.
- DNTPs are deoxyribose nucleotide triphosphates. These are the building blocks that are added during a PCR to synthesize a new strand. Examples are: dATP (adenosine triphosphate), dUTP (uracyl), dCTP (cytidine), dTTP (thymidine) and dGTP (guanosine).
- dATP adenosine triphosphate
- dUTP uracyl
- dCTP cytidine
- dTTP thymidine
- dGTP guanosine
- buffer stands for reaction buffer. The person skilled in the corresponding reaction buffer known.
- Polyethylene filter disks were used as porous supports.
- FIG. 1 On different zones of the filter disk, the following components were added (FIG. 1):
- FIG. 1 shows a filter disk with different zones and PCR materials.
- the components were applied in the amount corresponding to a ⁇ PCR approach.
- the solutions were placed on the filter disk and dried for about 2 h under physiological temperature conditions (37 ° C). After drying, the filter discs were transferred to a 2 ml screw, sealed with a lid and stored at room temperature for 3 months and tested after this storage time in a comparison reaction with freshly added reaction components.
- the DNA buffer mixture was added to the filter plate in the 2 ml tube and vortexed for 1 min.
- the 100 ⁇ each of the PCR mixture was distributed to 6 wells of a SpeedCycler plate of 15 ⁇ each. After carrying out the amplification reaction, the amplification products were evaluated on an agarose gel (FIG. 2).
- FIG. 2 shows PCR products after amplification of genomic DNA with fresh and storage-stable reagents.
- the compartmentalized reagents were prepared for the performance of a probe-based LFA assay.
- the compartmentalized transfer of the individual reaction components should prevent an irregular start of reaction and thus prevent the occurrence of false-positive results.
- Polyethylene filter disks were used as porous supports.
- FIG. 3 On different zones of the filter disk, the following components were added (FIG. 3):
- FIG. 3 shows the top view of a filter disk. Compartmentation of storage-stable components for a probe-based LFA assay.
- the reagents were applied in the amount corresponding to a 100 ⁇ PCR approach.
- the solutions were pipetted onto the filter disc and dried for approximately 2 h under physiological temperature conditions of 37.degree. After drying, the filter discs were placed in a 2 ml screw, sealed with a lid provided and stored at RT for 3 months and tested in a comparison reaction with freshly added reaction components.
- 2x15 ⁇ of the batch were pipetted off as a negative control, to the remainder of the master mix 5 ⁇ S. Enterica DNA were added and of which 2x15 ⁇ of the PCR mixture were added to the well of a PCR plate for the SpeedCycler.
- the DNA buffer mixture was placed on the filter disc in the 2 ml tube, capped and vortexed for 1 min. 2x15 ⁇ of the batch were pipetted off as a negative control, 5 ⁇ S. enterica DNA was added to the remainder of the master mix, and 2 ⁇ 15 ⁇ of the PCR mixture were added to the well of a PCR plate for the SpeedCycler. 3) "Fresh + Primer / Probe Mix"
- primer and probe were mixed together in the given amount (see “Frisch” approach) 2 hours prior to the reaction and stored at room temperature to favor dimer formation 2x15 ⁇ of the batch was pipetted off as a negative control to which The remainder of the master mix was added to 5 ⁇ S. enterica DNA and 2x15 ⁇ of the PCR mix was added to the well of a PCR plate for the SpeedCycler.
- Figure 4 shows an amlification hybridization reaction (RAH technology) after evaluation of the PCR approach on the gel (A) and on an LFA (B).
- RH technology amlification hybridization reaction
- Embodiment 3 Preparation of a compartmentalized storage-stable reaction mixture and use thereof for OneStep cDNA synthesis and amplification of an influenza A virus.
- Polyethylene filter disks were used as porous supports.
- FIG. 5 shows the top view of a filter disk. Compartmentation of storage-stable components for a OneStep PCR.
- buffer containing dNTPs was dried on the bottom of the reaction vessel (Herculase® II RT-PCR 2x Mastermix, Agilent).
- the chemicals were applied in the amount corresponding to a ⁇ approach.
- the solutions were placed on the filter disc or on the bottom of a 2 ml screw, screwed with a lid and dried for about 2 h at physiological temperature conditions of 37 ° C.
- the filter discs were placed in a 2 ml screw and stored at RT for 3 months and tested a comparative reaction with freshly added reaction components.
- RNA samples [0069] A dilution series of RNA samples from cultured influenza A viruses was prepared. The samples were diluted so that 12 ⁇ RNA each corresponded to 10 5 , 10 4 , 10 3 , 10 2 and 10 1 virus particles.
- RNA solution was placed on the filter plate in the 2 ml tube and vortexed for 1 min.
- FIG. 6 shows different virus dilutions were amplified with freshly prepared components (fresh) or storage-stable components (dried) on a filter disc in a OneStep RT-PCR and detected on a lateral flow strip.
- the example illustrates that the use of the storage-stable coated filter disk even with a OneStep RT-PCR reaction shows no loss of activity compared to freshly added reaction components.
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Analytical Chemistry (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Pathology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE102010038330A DE102010038330A1 (de) | 2010-07-23 | 2010-07-23 | Verfahren, Vorrichtung und Testkit für Molekularbiologische Reaktionen |
PCT/EP2011/062692 WO2012010708A1 (de) | 2010-07-23 | 2011-07-22 | Verfahren, vorrichtung und testkit für molekularbiologische reaktionen |
Publications (1)
Publication Number | Publication Date |
---|---|
EP2596129A1 true EP2596129A1 (de) | 2013-05-29 |
Family
ID=44545680
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP11751837.3A Withdrawn EP2596129A1 (de) | 2010-07-23 | 2011-07-22 | Verfahren, vorrichtung und testkit für molekularbiologische reaktionen |
Country Status (4)
Country | Link |
---|---|
US (1) | US20130280695A1 (de) |
EP (1) | EP2596129A1 (de) |
DE (1) | DE102010038330A1 (de) |
WO (1) | WO2012010708A1 (de) |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2013005796A (ja) | 2011-05-26 | 2013-01-10 | Arkray Inc | 乾燥試薬、乾燥試薬キット、試薬器、乾燥試薬の製造方法 |
RU2535333C1 (ru) * | 2013-05-31 | 2014-12-10 | Мераб Георгиевич Чикобава | Устройство для полимеразной цепной реакции |
US9705470B1 (en) | 2014-02-09 | 2017-07-11 | Sitime Corporation | Temperature-engineered MEMS resonator |
DE102016101948A1 (de) * | 2016-02-04 | 2017-08-10 | Leibniz-Institut Für Pflanzenbiochemie (Ipb) | Verfahren, Träger und Kit zur Ein-Topf-Ein-Schritt-Assemblierung von DNA-Molekülen |
CN111684077A (zh) | 2018-02-06 | 2020-09-18 | 简·探针公司 | 远红染料探针调配物 |
Family Cites Families (25)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE1220083B (de) * | 1965-02-16 | 1966-06-30 | Rudolf Siegert Dr | Vorrichtung zur Pruefung biochemischer Reaktionen an Bakterien |
US3645853A (en) * | 1969-06-24 | 1972-02-29 | Warner Lambert Co | Diagnostic composition and method for the detection of nitrate reduction |
GB1544995A (en) * | 1977-01-11 | 1979-04-25 | Oxoid Ltd | Gas-producing device |
US4891319A (en) | 1985-07-09 | 1990-01-02 | Quadrant Bioresources Limited | Protection of proteins and the like |
DE3738721C2 (de) | 1987-11-14 | 1995-10-05 | Roehm Gmbh | Immobilisierte Antikörper |
EP0511559A1 (de) | 1991-04-30 | 1992-11-04 | F.Hoffmann-La Roche & Co. Aktiengesellschaft | Reagens für eine Oligonukleotidprobe |
US5725831A (en) * | 1994-03-14 | 1998-03-10 | Becton Dickinson And Company | Nucleic acid amplification apparatus |
US7253262B2 (en) | 1995-01-19 | 2007-08-07 | Quandrant Drug Delivery Limited | Dried blood factor composition comprising trehalose |
US5556771A (en) * | 1995-02-10 | 1996-09-17 | Gen-Probe Incorporated | Stabilized compositions of reverse transcriptase and RNA polymerase for nucleic acid amplification |
US6319676B1 (en) | 1995-05-02 | 2001-11-20 | Carter Wallace, Inc. | Diagnostic detection device and method |
CN1095693C (zh) * | 1996-09-16 | 2002-12-11 | 阿尔法海利克斯股份公司 | 存储和发放试剂用的药筒和方法 |
US5861251A (en) * | 1996-10-15 | 1999-01-19 | Bioneer Corporation | Lyophilized reagent for polymerase chain reaction |
DE19647055A1 (de) | 1996-11-14 | 1998-05-20 | Boehringer Mannheim Gmbh | Stabilisierte wässrige Nukleosidtriphosphat-Lösungen |
US5905029A (en) * | 1997-02-19 | 1999-05-18 | Fritz Berthold | Method for rapid hygiene testing |
US5939259A (en) * | 1997-04-09 | 1999-08-17 | Schleicher & Schuell, Inc. | Methods and devices for collecting and storing clinical samples for genetic analysis |
AU2001262483A1 (en) * | 2000-05-31 | 2001-12-11 | Tepnel Medical Limited | Formulation for polymerase chain reaction and vessel containing same |
DE10053553C2 (de) | 2000-10-28 | 2002-09-19 | Bag Biolog Analysensystem Gmbh | Vorrichtung zum Nachweis und zum Anreichern von Biomolekülen |
US20020197631A1 (en) * | 2001-04-26 | 2002-12-26 | Lawrence Nathan P. | Multichamber device and uses thereof for processing of biological samples |
FR2855177B1 (fr) | 2003-05-23 | 2005-07-08 | Raymond Robert | Composition contenant des acides nucleiques lyophilises en presence de collagene et de disaccharides |
US20050069898A1 (en) | 2003-09-25 | 2005-03-31 | Cepheid | Lyophilized beads containing mannitol |
DE102004021822B3 (de) | 2004-04-30 | 2005-11-17 | Siemens Ag | Verfahren und Anordnung zur DNA-Amplifikation mittels PCR unter Einsatz von Trockenreagenzien |
FI20040768A0 (fi) * | 2004-06-04 | 2004-06-04 | Teemu Korpimaeki | Menetelmä määritysreagenssien stabiloimiseksi, stabilisoituja määritysreagensseja sisältävä reagenssisäiliö ja sen käyttö |
ES2401879T3 (es) * | 2004-08-31 | 2013-04-25 | Eiken Kagaku Kabushiki Kaisha | Método de análisis de ácidos nucleicos |
EP2089543B1 (de) * | 2006-11-15 | 2020-07-01 | BioFire Diagnostics, LLC | Hochdichtes und eigenständiges biologisches analyseverfahren |
DE102006056790B3 (de) | 2006-12-01 | 2008-01-17 | IfP Privates Institut für Produktqualität GmbH | Laboreinmalartikel für Analyse und Diagnosik |
-
2010
- 2010-07-23 DE DE102010038330A patent/DE102010038330A1/de not_active Withdrawn
-
2011
- 2011-07-22 WO PCT/EP2011/062692 patent/WO2012010708A1/de active Application Filing
- 2011-07-22 EP EP11751837.3A patent/EP2596129A1/de not_active Withdrawn
- 2011-07-22 US US13/811,841 patent/US20130280695A1/en not_active Abandoned
Non-Patent Citations (2)
Title |
---|
None * |
See also references of WO2012010708A1 * |
Also Published As
Publication number | Publication date |
---|---|
DE102010038330A1 (de) | 2012-03-01 |
WO2012010708A1 (de) | 2012-01-26 |
US20130280695A1 (en) | 2013-10-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP7009011B2 (ja) | 直接核酸増幅キット、試薬及び方法 | |
EP2430195B1 (de) | Zusammensetzungen und verfahren zur lagerung biologischer proben | |
Wendel | Assessing viability and stress tolerance of probiotics—a review | |
JP3087907B2 (ja) | ポリメラーゼチェイン反応の特異性と簡便性の改良 | |
DE69628959T2 (de) | Stabilisierte Zusammensetzungen von Reverser Transkriptase und RNA Polymerase für Nukleinsäureamplifikation | |
DE69819124T2 (de) | Lyophilisation von kultivierten, menschlichen zellen zur konservierung der rna und dna | |
AU2005258951B2 (en) | Method for stabilising reagents which are useful for nucleic acid amplification | |
DE102017201810B4 (de) | Getrocknete amplifikationszusammensetzungen | |
EP3548895B1 (de) | Aufbereitung lebendiger, mikrobieller proben und mikroorganismen für anschliessende massenspektrometrische messung und auswertung | |
DE102006056790B3 (de) | Laboreinmalartikel für Analyse und Diagnosik | |
US20120100522A1 (en) | Stabilized chemical dehydration of biological material | |
WO2012010708A1 (de) | Verfahren, vorrichtung und testkit für molekularbiologische reaktionen | |
US10590408B2 (en) | Method and composition for biomolecule stabilization | |
US20120064536A1 (en) | Dried and Stabilized Ready-to-Use Composition Containing Nucleic Acid Polymerization Enzymes for Molecular Biology Applications | |
DE112018002597B4 (de) | Getrocknete zusammensetzungen, die flap-endonuklease enthalten | |
DE102020103957B4 (de) | Verfahren zum Nachweis von Mikroorganismen und ein fluidisches Kanalsystem | |
DE2711754C2 (de) | Verfahren zum Stabilisieren eines Enzyms, das in wäßrigem Medium instabil ist | |
RU171152U1 (ru) | Дозированная форма для полимеразной цепной реакции | |
WO2013120853A1 (de) | Anordnung und verfahren zum nachweis von mikroorganismen in einem kulturgefäss | |
DE102012014981B3 (de) | Verfahren zur Differenzierung zwischen lebenden und toten Zellen | |
DE102009054098A1 (de) | Nachweis von Mykobakterien und deren Resistenzgenen |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 20130225 |
|
AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR |
|
RIN1 | Information on inventor provided before grant (corrected) |
Inventor name: HILLEBRAND, TIMO Inventor name: DASKALOW, KATJANA Inventor name: GRASER, ELMARA |
|
DAX | Request for extension of the european patent (deleted) | ||
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: EXAMINATION IS IN PROGRESS |
|
17Q | First examination report despatched |
Effective date: 20171109 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: EXAMINATION IS IN PROGRESS |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN |
|
18D | Application deemed to be withdrawn |
Effective date: 20220201 |