CN105969899A - Method and device for enriching trace target nucleic acid based on perforated plate - Google Patents
Method and device for enriching trace target nucleic acid based on perforated plate Download PDFInfo
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- CN105969899A CN105969899A CN201610580044.5A CN201610580044A CN105969899A CN 105969899 A CN105969899 A CN 105969899A CN 201610580044 A CN201610580044 A CN 201610580044A CN 105969899 A CN105969899 A CN 105969899A
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- 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
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Abstract
The invention relates to a method and device for enriching trace target nucleic acid based on a perforated plate. The method for enriching the trace target nucleic acid based on the perforated plate comprises the following steps: (1) injecting a certain amount of nucleic acid crude extract or a high-purity nucleic acid sample into reaction holes of the perforated plate; (2) absorbing the nucleic acid crude extract or high-purity nucleic acid sample in one reaction hole of the perforated plate by using magnetic particles, and then removing liquid; (3) quantitatively transferring the nucleic acid crude extract or high-purity nucleic acid sample in other reaction holes into the reaction hole used in the step (2), absorbing the nucleic acid crude extract or high-purity nucleic acid sample by using the magnetic particles, and then removing liquid; (4) repeating the steps (2) and (3) until the amount of the trace target nucleic acid absorbed by the magnetic particles reaches an analyzable amount; and (5) eluting the nucleic acid absorbed on the magnetic particles. The method and the device take the perforated plate as a carrier and are capable of automatically completing the enrichment for the trace target nucleic acid, thus enabling full-automatic detection of automatic microbial molecular detection equipment for trace samples to become possible.
Description
Technical field
The invention belongs to mensuration or the method for inspection technical field of microorganism, particularly to a kind of base
The enrichment method of trace target nucleic acids and device in porous plate.
Background technology
It is conventional nucleic acid purification method with magnetic particle absorption nucleic acid, such as Application No.
200980123037.0 innovation and creation disclosed in a kind of side of purifying nucleic acid from fixing organization
Method mainly have employed by magnetic particle absorption the thought of eluting nucleic acid.
For needing the sample processed in batch, porous plate is a kind of conventional carrier.Application number
Be 201380018553.3 innovation and creation disclose a kind of for automatically analyzing Biosample
Equipment and method, it have employed porous plate test kit, a porous plate test kit when purification
The amount of the sample to be tested being built into is the least, if the content of target nucleic acids is the highest, the most existing
Technology cannot use automation equipment to detect.That is: for trace sample, a porous plate
The target nucleic acids contained by nucleic acid cleavage liquid in reacting hole does not reaches the content being detected, existing skill
Automatic detection system in art not yet has solution.This becomes automatization's microbial molecules
The bottleneck of detection equipment development.
Summary of the invention
It is an object of the invention to overcome the deficiencies in the prior art, it is provided that a kind of trace based on porous plate
The enrichment method of amount target nucleic acids, makes the trace sample being contained in porous plate to conveniently realize
The enrichment of nucleic acid.
The present invention solves the scheme of described technical problem: a kind of trace target based on porous plate
The enrichment method of nucleic acid, comprises the steps:
(1) in the reacting hole of porous plate, a certain amount of nucleic acid crude extract is injected or through purification
After high-purity nucleic acid sample of obtaining；
(2) the nucleic acid crude extract in the reacting hole of porous plate or high-purity nucleic acid sample are carried out magnetic
Property particle absorption operation, remove liquid afterwards；
(3) the nucleic acid crude extract in other reacting holes or high-purity nucleic acid sample are moved into quantitatively
In the reacting hole of step (2), after carrying out magnetic particle adsorption operations, remove liquid；
(4) step (2)-(3) are repeated, until the trace target nucleic acids of magnetic particle absorption
Till reaching analyzable amount；
(5) nucleic acid eluents is added, eluting absorption nucleic acid on magnetic particle.
As improvement, described nucleic acid cleavage liquid obtains through the following step that processes: (1) takes necessarily
Porous plate put into by the sample of quality or volume；(2) add cell pyrolysis liquid, carry out as required
Heating and/or oscillation treatment；(3) nucleic acid crude extract is obtained after filtering.
As improvement, described nucleic acid crude extract or through high-purity nucleic acid sample of obtaining after purification
Movement is carried out by multichannel liquid-transfering gun.
As improvement, the nucleic acid crude extract of same sample or through the high-purity core obtained after purification
Acid sample is positioned at same a line of same porous plate or with on string.
As improvement, the nucleic acid crude extract of same sample or through the high-purity core obtained after purification
Acid sample is positioned in the same position of different porous plate.
As improvement, the step of described magnetic particle adsorption operations is: magnetic particle is added by (1)
Enter the reacting hole to porous plate, and shake；(2) applying magnetic field around porous plate reacting hole；
(3) on the premise of magnetic field maintains magnetic particle, liquid is removed.
It is a further object of the present invention to provide a kind of device realizing said method, its technical scheme
For:
The enriching apparatus of a kind of trace target nucleic acids based on porous plate, including porous plate, magnetic field
Generator, the multichannel liquid-transfering gun controlled by mechanical arm, magnetic field generation device is in porous
Panel area produces magnetic field, mechanical arm the multichannel liquid-transfering gun controlled is for adding porous by liquid
In the reacting hole of plate or liquid is removed from the reacting hole of porous plate.
As improvement, described magnetic field generation device is electromagnetic type magnetic field generation device, and magnetic field produces
Depression for housing porous plate is arranged at the top of device.
As improvement, the quantity of described porous plate is more than 2 pieces, the same position of every block of porous plate
Place the nucleic acid crude extract of sample of the same race or through the high-purity nucleic acid sample obtained after purification.
The application is with porous plate as carrier, and its main thought is: by certain mass or the examination of volume
In sample is placed in the reacting hole of porous plate and make sample complete cracking to obtain nucleic acid cleavage liquid, nucleic acid splits
Solving liquid and obtain nucleic acid crude extract after filtering, nucleic acid crude extract carries through the absorption of magnetic particle
High-purity nucleic acid sample is obtained after pure.The object of the application enrichment both can be nucleic acid crude extract, also
It can be the high-purity nucleic acid sample obtained after purification.Same sample may be located at same porous
Same a line of plate or same string, same sample may be alternatively located at the same position of different porous plate,
To facilitate multichannel liquid-transfering gun to be moved in reacting hole added with magnetic particle by sample of the same race, make through
Cross the nucleic acid contained by magnetic particle after repeatedly absorption and reach detectable dosage.The application makes position
Nucleic acid in porous plate reacting hole is enriched with, and once can be to the nucleic acid of multiple samples
It is enriched with, improves work efficiency, make automatization of the prior art microbial molecules detection
Equipment carries out fully-automated synthesis to trace sample and is possibly realized.
Accompanying drawing explanation
Fig. 1 is the structural representation of device of the present invention.
In figure: 1, porous plate；2, magnetic field generation device；21, depression；3, mechanical arm；4、
Multichannel liquid-transfering gun.
Detailed description of the invention
Embodiment 1
The enrichment method of a kind of trace target nucleic acids based on porous plate, comprises the steps:
(1) multichannel liquid-transfering gun is used to inject a certain amount of nucleic acid in the reacting hole of porous plate
Crude extract or through the high-purity nucleic acid sample obtained after purification, the nucleic acid crude extract of same sample
Or the high-purity nucleic acid sample through obtaining after purification is positioned at same a line of same porous plate or same
On string, the nucleic acid crude extract of same sample or through the high-purity nucleic acid sample obtained after purification
May be alternatively located in the same position of different porous plate；
(2) to the nucleic acid crude extract in the reacting hole of porous plate or through the height obtained after purification
Pure nucleic acid sample carries out magnetic particle adsorption operations, uses multichannel liquid-transfering gun to remove liquid afterwards
Body；
(3) by the nucleic acid crude extract in other reacting holes or through the high-purity core obtained after purification
Acid sample uses multichannel liquid-transfering gun to move into quantitatively in the reacting hole of step (2), carries out magnetic
Property particle absorption operation after use multichannel liquid-transfering gun remove liquid；
(4) step (2)-(3) are repeated, until the trace target nucleic acids of magnetic particle absorption
Till reaching analyzable amount；
(5) nucleic acid eluents is added, eluting absorption nucleic acid on magnetic particle.
Described nucleic acid cleavage liquid obtains through the following step that processes: (1) takes certain mass or volume
Sample put into porous plate；(2) add cell pyrolysis liquid, carry out as required heating and/or
Oscillation treatment；(3) nucleic acid crude extract is obtained after filtering.
The step of described magnetic particle adsorption operations is: magnetic particle is added to porous plate by (1)
Reacting hole, and shake；(2) applying magnetic field around porous plate reacting hole；(3) at magnetic
Field uses multichannel liquid-transfering gun to remove liquid on the premise of maintaining magnetic particle.
The device corresponding with the method is as it is shown in figure 1, a kind of trace target core based on porous plate
The enriching apparatus of acid, including porous plate 1, magnetic field generation device 2, is controlled by mechanical arm 3
Multichannel liquid-transfering gun 4, porous plate 1 is positioned at when purifying nucleic acid on magnetic field generation device 2,
Magnetic field generation device 2, for producing magnetic field around porous plate 1, is controlled many by mechanical arm 3
Passage liquid-transfering gun 4 in the reacting hole that liquid is added porous plate 1 or by liquid from porous plate
The reacting hole of 1 removes.
Magnetic field generation device 2 is electromagnetic type magnetic field generation device, produces magnetic field, close after energising
After power supply, magnetic field disappears.Depression for housing porous plate 1 is arranged at the top of magnetic field generation device 2
21。
The quantity of porous plate 1 is more than 2 pieces, and the same position of every block of porous plate 1 places examination of the same race
The nucleic acid crude extract of sample or through the high-purity nucleic acid sample obtained after purification.During enrichment, more than one piece
Orifice plate 1 is positioned at the depression 21 of magnetic field generation device 2, and other porous plates 1 are positioned at operating board
On face, what all porous plates 1 were placed in the reacting hole of same position is the nucleic acid of same sample
Crude extract or through the high-purity nucleic acid sample obtained after purification.
Claims (9)
1. the enrichment method of a trace target nucleic acids based on porous plate, it is characterised in that
Comprise the steps:
(1) in the reacting hole of porous plate, a certain amount of nucleic acid crude extract is injected or through purification
After high-purity nucleic acid sample of obtaining；
(2) the nucleic acid crude extract in the reacting hole of porous plate or high-purity nucleic acid sample are carried out magnetic
Property particle absorption operation, remove liquid afterwards；
(3) the nucleic acid crude extract in other reacting holes or high-purity nucleic acid sample are moved into quantitatively
In the reacting hole of step (2), after carrying out magnetic particle adsorption operations, remove liquid；
(4) step (2)-(3) are repeated, until the trace target nucleic acids of magnetic particle absorption
Till reaching analyzable amount；
(5) nucleic acid eluents is added, eluting absorption nucleic acid on magnetic particle.
2. the enrichment method of trace target nucleic acids based on porous plate as claimed in claim 1,
It is characterized in that, described nucleic acid crude extract obtains through the following step that processes: (1) takes certain matter
Porous plate put into by the sample of amount or volume；(2) add cell pyrolysis liquid, add as required
Heat and/or oscillation treatment；(3) nucleic acid crude extract is obtained after filtering.
3. the enrichment method of trace target nucleic acids based on porous plate as claimed in claim 1,
It is characterized in that, described nucleic acid crude extract or the shifting of the high-purity nucleic acid sample through obtaining after purification
Fortune is carried out by multichannel liquid-transfering gun.
4. the enrichment method of trace target nucleic acids based on porous plate as claimed in claim 1,
It is characterized in that, the nucleic acid crude extract of same sample or through the high-purity nucleic acid obtained after purification
Sample is positioned at same a line of same porous plate or with on string.
5. the enrichment method of trace target nucleic acids based on porous plate as claimed in claim 1,
It is characterized in that, the nucleic acid crude extract of same sample or through the high-purity nucleic acid obtained after purification
Sample is positioned in the same position of different porous plate.
6. the enrichment method of trace target nucleic acids based on porous plate as claimed in claim 1,
It is characterized in that, the step of described magnetic particle adsorption operations is: magnetic particle is added by (1)
To the reacting hole of porous plate, and shake；(2) applying magnetic field around porous plate reacting hole；
(3) on the premise of magnetic field maintains magnetic particle, liquid is removed.
7. the enriching apparatus of a trace target nucleic acids based on porous plate, it is characterised in that
Including porous plate (1), magnetic field generation device (2), the multichannel that controlled by mechanical arm (3)
Liquid-transfering gun (4), magnetic field generation device (2) is used for producing magnetic field around porous plate (1),
The multichannel liquid-transfering gun (4) controlled by mechanical arm (3) for adding porous plate (1) by liquid
Reacting hole in or liquid is removed from the reacting hole of porous plate (1).
8. the enriching apparatus of trace target nucleic acids based on porous plate as claimed in claim 7,
It is characterized in that, described magnetic field generation device (2) is electromagnetic type magnetic field generation device, magnetic field
Depression (21) for housing porous plate (1) is arranged at the top of generator (2).
9. the enriching apparatus of trace target nucleic acids based on porous plate as claimed in claim 7,
It is characterized in that, the quantity of described porous plate (1) is more than 2 pieces, every piece of porous plate (1)
Same position places the nucleic acid crude extract of sample of the same race or through the high-purity nucleic acid obtained after purification
Sample.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610580044.5A CN105969899A (en) | 2016-07-19 | 2016-07-19 | Method and device for enriching trace target nucleic acid based on perforated plate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610580044.5A CN105969899A (en) | 2016-07-19 | 2016-07-19 | Method and device for enriching trace target nucleic acid based on perforated plate |
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| CN105969899A true CN105969899A (en) | 2016-09-28 |
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| CN201610580044.5A Pending CN105969899A (en) | 2016-07-19 | 2016-07-19 | Method and device for enriching trace target nucleic acid based on perforated plate |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102417902A (en) * | 2011-09-16 | 2012-04-18 | 广西出入境检验检疫局检验检疫技术中心 | Kit for extracting nucleic acid by magnetic bead-microtiter plate method and method for extracting nucleic acid by same |
| CN103282782A (en) * | 2010-10-27 | 2013-09-04 | 株式会社百奥尼 | Automatic real-ime pcr system for the various analysis of biological smaple |
| CN104204229A (en) * | 2012-02-10 | 2014-12-10 | 株式会社百奥尼 | Apparatus and method for automatically analyzing biological samples |
-
2016
- 2016-07-19 CN CN201610580044.5A patent/CN105969899A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103282782A (en) * | 2010-10-27 | 2013-09-04 | 株式会社百奥尼 | Automatic real-ime pcr system for the various analysis of biological smaple |
| CN102417902A (en) * | 2011-09-16 | 2012-04-18 | 广西出入境检验检疫局检验检疫技术中心 | Kit for extracting nucleic acid by magnetic bead-microtiter plate method and method for extracting nucleic acid by same |
| CN104204229A (en) * | 2012-02-10 | 2014-12-10 | 株式会社百奥尼 | Apparatus and method for automatically analyzing biological samples |
Non-Patent Citations (1)
| Title |
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| 罗英: "磁珠法核酸自动提取仪在分子生物学领域的应用", 《蚕学通讯》 * |
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| TA01 | Transfer of patent application right |
Effective date of registration: 20200825 Address after: 1213-177, 12 / F, building 3, No. 1366, Hongfeng Road, Kangshan street, Huzhou Economic and Technological Development Zone, Huzhou City, Zhejiang Province Applicant after: Original microbial detection technology (Huzhou) Co.,Ltd. Address before: 313008, room two, building 1888, building E, No. 003 Dai Shan Road, Wuxing District, Zhejiang, Huzhou, China Applicant before: HUZHOU HIGH-INNO BIOTECHNOLOGY Co.,Ltd. |
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Application publication date: 20160928 |