CN106978333A - Genetic test and parting based on microwell plate - Google Patents
Genetic test and parting based on microwell plate Download PDFInfo
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- CN106978333A CN106978333A CN201611196221.6A CN201611196221A CN106978333A CN 106978333 A CN106978333 A CN 106978333A CN 201611196221 A CN201611196221 A CN 201611196221A CN 106978333 A CN106978333 A CN 106978333A
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- 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
- C12Q1/6837—Enzymatic or biochemical coupling of nucleic acids to a solid phase using probe arrays or probe chips
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
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Abstract
The present invention relates to a kind of device, the multi-well microplate device of especially a kind of high flux genetic test and parting；The device of the patent is made up of multi-well microplate and the genetic chip that is embedded in micropore, and multi-well microplate can be various passes, the microwell plate of different hole counts, including circle, square or other passes 12 holes, 24 holes, 48 holes, 96 holes, 384 holes etc., the genetic chip being embedded in micropore can also be with nylon membrane, silicon chip, sheet glass or sheet are matrix, contain the specialized probe arrays for Genotyping；The present invention has reformed the single means of traditional genetic test, fully combining genetic chip once can be while detects the advantage of multiple genes and the advantage that can be once detected using multi-well microplate to multiple sample in a sample, genetic chip is embedded in multi-well microplate and is configured to high pass amount detecting device, with simple to operate, time saving and energy saving province's reagent, cross pollution is reduced, detection is accurate, the advantages of as a result judging directly perceived.
Description
Technical field
This patent is related to a kind of device, is based particularly on genetic test and the parting of microwell plate, belongs to biological technical field.
Background technology
Genetic chip is to hybridize using known nucleic acid sequence as probe with complementary target nucleotide sequences, by subsequent
Signal detection carries out quantification and qualification.Genetic chip is integrated with substantial amounts of molecular recognition in a small substrate surface and visited
Pin, can within the same time the substantial amounts of gene of parallel analysis, carry out large information capacity screening and detection and analysis.Current clinic
Detection is much applied to biochip technology with gene diagnosis, can be quickly detected from the table of thousands of genes in sample
Up to situation.Although genetic chip can same time parallel analysis lots of genes, up to the present, the base of commercialization on the market
Because detection chip is substantially the design for a person-portion, testing staff is also only limited to every time with a genechip detection portion
The expression conditions of sample, often detect a sample, the step needed for detection is performed one time must again again, time-consuming to take
Power, and because the complicated step of multiple sample detections repeats to cause to have when muscle power and spirit consumption, failure and the detection of mistake
Occur.So far multiple sample can also be detected simultaneously with lots of genes chip without extraordinary product on the market.
Patent content
The purpose of this patent is one device of structure, can carry out genetic test and parting to multiple sample with high throughput,
Time saving and energy saving province's reagent is reached, accurate purpose is detected.
This patent is characterized by, and the device is made up of multi-well microplate and the genetic chip that is embedded in micropore, is embedded in micro-
Genetic chip in hole, contains the specialized probe arrays for genetic test and parting；The genetic chip can with nylon membrane,
Silicon chip, sheet glass, sheet is matrix.Preferred substrate is nylon membrane, it is easy to cut, and cost is low, simple to operate, as a result background
It is low.Multi-well microplate can be various passes, the microwell plate of different hole counts, including circle, square or other passes 12 holes, 24
Hole, 48 holes, 96 holes, 384 holes etc.；Microwell plate selection can according to it is required detection genotype number depending on；According to what is used
Film quality chip, can be cut into correspondingly sized and embedded micropore by the micropore size of different shaped jet holes or the microwell plate of different hole counts,
It is built into the genetic chip device of three-dimensional.In this device, microwell plate also serves as gene inspection both as the carrier of multiple chips
The reaction vessel of survey.
This patent has reformed the single means of traditional genetic test, by the genetic chip and microwell plate of existing two-dimensional structure
With reference to the device for being built into three-dimensional, fully combining genetic chip once can be while detects the advantage of multiple genes in a sample
With the advantage that can be once detected using multi-well microplate to multiple sample, genetic chip is embedded in multi-well microplate and constructed
Into high pass amount detecting device, with simple to operate, time saving and energy saving province's reagent, cross pollution is reduced, detection is accurate, as a result judge straight
The advantages of sight.
Brief description of the drawings
Fig. 1 patent device schematic diagrames.
Fig. 2 left figures：The arrangement of genetic matrix, right figure on genetic chip：Minute of individual gene matrix middle probe and Quality Control point
Cloth (including different genotype probe ,+ve:Position Quality Control point ,-ve:：Negative Quality Control point, HB：Sample DNA is extracted and PCR processes
Positive quality control point).
Embodiment
The device of this patent is described further with reference to the drawings and specific embodiments, but does not limit this in any form
Patent：
This example of the structure of the microplate apparatus of the high flux genetic test of embodiment one and parting is hardened using 96 square hole micropores
It is the genetic chip that 6.5 millimeters of square nylon membranes are matrix to close the length of side.
Nylon membrane is cut into suitable size, is fixed on point sample instrument, and according to the size of required matrix, point sample instrument can be once
Complete gene matrix 384-500 pieces.The number of probes of design is simultaneously arranged into matrix, sets full-automatic in point sample instrument
Program carries out point sample.For the ease of the fixation of probe, probe has carried out poly T at 5 ends and amido modified, and in the work of catalyst
Under, it is fixed on by covalent bond on nylon membrane, other sites will be inactivated with chemical action, gene is formed on genetic chip 3
Matrix 4.
By being chemically treated and the genetic chip 3 that finishes of point sample just can have been used, be cut into 6.5 millimeters it is square simultaneously
In the micropore 2 for being put into 96 square hole microwell plates 1.The microplate apparatus of high flux genetic test and parting just completes to build, and it is illustrated
Figure can be found in Fig. 1.The reaction reagent consumption of each micropore 2 only needs 100-200 microlitres, and this microplate apparatus can detect 96 people simultaneously
Part.
Application of the device that embodiment two is built in the genetic test and parting of human papilloma virus in this instance,
The device form of embodiment one is continued to use, in genetic test and parting that this device is applied to human papilloma virus.
There are 100 Multi-genotypes, and kind has close relationship with cervix cancer and condyloma acuminatum wherein more than 40, is selected in this example
29 kinds of common genotype are taken, including are 6,11,16,18,26,31,33,35,39,40,42,43,44,45,51,52,53,
54,55,56,57,58,59,66,67,68,69,73,82.
The design alternative of probe is the variable region positioned at HPV L1 regions, specificity spy of the design for 29 kinds of genotype
Pin.This genetic chip, specific probe is arranged as 6x6 matrixes with Quality Control point on nylon membrane, as shown in Fig. 2 left figures, matrix dot
At intervals of 600 microns between point, the distribution of its specific specific probe and Quality Control point is as shown in Fig. 2 right figures.
HPV micropore plate hybridization testing process combination PCRs, target dna is expanded with multiple.In HPV genes
The L1 regions design universal primer of group, primer is with biotin labeling, after amplification, and the PCR primer with biotin labeling passes through alkali
Denaturation, can be captured after neutralization by the corresponding specific probe being fixed on genetic chip.After the completion of hybridization, pass through peroxide
The Streptavidin of compound enzyme mark and its specific substrate TMB (TMB) chromogenic reaction are drawn
As a result.IMAQ and analysis are carried out to gene chip results, gene type data is provided in a short time.
96 parts of different samples can be detected simultaneously using the device in this, with utilizing traditional parting hybridizing method
(Roche HPV parting kits) equally 96 parts of samples of detection, take, reagent dosage, the comparison made in terms of accuracy in operation
It is as shown in the table：
| Comparison other | Operating time (hour) | Reagent dosage (%) | Detection unsuccessfully counts (part) |
| The present invention | 2.5 | 100 | 1 |
| Roche HPV parting kits | At least 24 | 5000 | 7 |
Claims (6)
1. genetic test and parting device based on microwell plate, it is characterised in that the device is by multi-well microplate and is embedded in micropore
Genetic chip composition.
2. high flux genetic test as claimed in claim 1 and the microplate apparatus of parting, it is characterised in that described is porous
Microwell plate can be square hole plate or circular hole slab or the plate of other passes.
3. high flux genetic test as claimed in claim 1 or 2 and the microplate apparatus of parting, it is characterised in that described is more
Hole microwell plate can be 12 holes or 24 holes or 48 holes or 96 holes or 384 hole microwell plates.
4. high flux genetic test as claimed in claim 1 and the microplate apparatus of parting, it is characterised in that described gene
Chip can be using nylon membrane as matrix, or using silicon chip as matrix, or using sheet glass as matrix, or using sheet as matrix.
5. high flux genetic test as claimed in claim 1 and the microplate apparatus of parting, it is characterised in that described gene
Chip is using nylon membrane as matrix.
6. high flux genetic test as claimed in claim 1 and the microplate apparatus of parting, it is characterised in that described gene
Chip contains the specialized probe arrays for genetic analysis and parting.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611196221.6A CN106978333A (en) | 2016-12-22 | 2016-12-22 | Genetic test and parting based on microwell plate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611196221.6A CN106978333A (en) | 2016-12-22 | 2016-12-22 | Genetic test and parting based on microwell plate |
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| Publication Number | Publication Date |
|---|---|
| CN106978333A true CN106978333A (en) | 2017-07-25 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611196221.6A Pending CN106978333A (en) | 2016-12-22 | 2016-12-22 | Genetic test and parting based on microwell plate |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110079450A (en) * | 2019-05-10 | 2019-08-02 | 广州安必平医药科技股份有限公司 | It is a kind of for the solid phase carrier chip system of in situ hybridization and its application |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN201280550Y (en) * | 2008-05-15 | 2009-07-29 | 港龙生物技术（深圳）有限公司 | Micro-pore panel apparatus for high flux gene detection and parting |
-
2016
- 2016-12-22 CN CN201611196221.6A patent/CN106978333A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN201280550Y (en) * | 2008-05-15 | 2009-07-29 | 港龙生物技术（深圳）有限公司 | Micro-pore panel apparatus for high flux gene detection and parting |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110079450A (en) * | 2019-05-10 | 2019-08-02 | 广州安必平医药科技股份有限公司 | It is a kind of for the solid phase carrier chip system of in situ hybridization and its application |
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Application publication date: 20170725 |