CN203569116U - Multiple (Polymerase Chain Reaction) detection kit - Google Patents
Multiple (Polymerase Chain Reaction) detection kit Download PDFInfo
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- CN203569116U CN203569116U CN201320686173.4U CN201320686173U CN203569116U CN 203569116 U CN203569116 U CN 203569116U CN 201320686173 U CN201320686173 U CN 201320686173U CN 203569116 U CN203569116 U CN 203569116U
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- 238000001514 detection method Methods 0.000 title claims abstract description 47
- 238000003752 polymerase chain reaction Methods 0.000 title abstract description 4
- 239000013642 negative control Substances 0.000 claims abstract description 53
- 239000013641 positive control Substances 0.000 claims abstract description 52
- 239000011148 porous material Substances 0.000 claims abstract description 38
- 238000006243 chemical reaction Methods 0.000 claims description 37
- 230000003321 amplification Effects 0.000 claims description 24
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 24
- 239000012745 toughening agent Substances 0.000 claims description 24
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- 238000012360 testing method Methods 0.000 description 15
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- 238000000034 method Methods 0.000 description 7
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- 102000039446 nucleic acids Human genes 0.000 description 3
- 238000003753 real-time PCR Methods 0.000 description 3
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- 206010011831 Cytomegalovirus infection Diseases 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 2
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- 241000282414 Homo sapiens Species 0.000 description 2
- 241000589248 Legionella Species 0.000 description 2
- 208000007764 Legionnaires' Disease Diseases 0.000 description 2
- 241000351643 Metapneumovirus Species 0.000 description 2
- 241000202934 Mycoplasma pneumoniae Species 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- 241000725643 Respiratory syncytial virus Species 0.000 description 2
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Images
Abstract
The utility model provides a multiple (Polymerase Chain Reaction) detection kit which is applicable to the technical field of biomedicine. The multiple (Polymerase Chain Reaction) detection kit comprises a kit body, a kit cover, a supporting seat, a negative control liquid tube, a positive control liquid tube and a 96-pore PCR plate; the supporting seat is located above the kit body; a negative control liquid tube hole, a positive control liquid tube hole and a 96-pore PCR plate hole are formed in the supporting seat, wherein the negative control liquid pipe hole and the positive control liquid pipe hole are formed in the same side of the 96-pore PCR plate hole; the negative control liquid tube, the positive control liquid tube and the 96-pore PCR plate hole are separably positioned in the negative control liquid pipe hole, the positive control liquid pipe hole and the 96-pore PCR plate hole, respectively. The multiple detection kit has the advantages of short cycle and multiple detections, and avoids pollution, etc.
Description
Technical field
The utility model belongs to field of biomedicine technology, relates in particular to a kind of multiple PCR detection kit.
Background technology
PCR(polymerase chain reaction) detection technique is the conventional detection means of molecular biology, be generally used for detecting the existence of certain section of object nucleotide sequence, and fluorescence quantitative PCR detection is due to highly sensitive, sense cycle is short, level of automation height etc., and advantage is widely used.Fluorescence quantitative PCR detection utilizes quantitative real time PCR Instrument to carry out conventionally, and various reaction reagents are mixed in PCR reaction tubes, then PCR reaction tubes is placed in to quantitative real time PCR Instrument and starts reaction.Step the most loaded down with trivial details in PCR testing process is mixed various samples exactly in PCR reaction tubes, because will add PCR damping fluid, archaeal dna polymerase, primer, MgCl
2, dNTP and water etc., and these reagent are conventionally in dispersion state, these reagent of buying are all separately to store.For fear of reagent contamination, while adding different reagent, all to change liquid transfer gun head at every turn, the loss of these rifle heads has also increased testing cost.This application of sample flow process has consumed the time has increased testing cost.Be necessary thus to provide a kind of detection kit that testing process reduces testing cost simultaneously that shortens.
Utility model content
The purpose of this utility model is to provide a kind of multiple PCR detection kit, and being intended to solve in prior art multiple fluorescence quantitative PCR, to detect in operation the application of sample cycle long, the problem more than liquid transfer gun head loss.
The utility model is to realize like this, a kind of multiple PCR detection kit, comprise box body, lid, supporting seat, negative controls pipe, positive control solution pipe, 96 hole PCR plates, this supporting seat is positioned at this box body top, on this supporting seat, be provided with negative controls pore, positive control solution pore, 96 hole PCR plate holes, this negative controls pore and positive control solution pore are positioned at 96 PCR plate hole the same sides, hole, and this negative controls pipe, positive control solution pipe and 96 hole PCR plates are positioned at respectively this negative controls pore separably, in positive control solution pore and 96 hole PCR plate holes.
Particularly, be formed in one structure or demountable structure of the structure of this 96 hole PCR plate.
Particularly, described demountable structure is comprised of support and 12 eight unions.
Particularly, this multiple PCR detection kit also comprises amplification toughener pipe, and on this supporting seat, be provided with amplification toughener pore, this amplification toughener pore and this negative controls pore are positioned at 96 PCR plate hole the same sides, hole, and this amplification toughener pipe is arranged in this amplification toughener pore separably.
Particularly, this 96 hole PCR plate comprises 96 PCR reaction tubess, in this PCR reaction tubes, PCR reaction solution is all housed.
Particularly, in this negative controls pipe, negative controls is housed, in positive control solution pipe, positive control solution is housed.
Particularly, this negative controls pipe is different from this positive control solution pipe specification.
Particularly, in this amplification toughener pipe, amplification toughener is housed.
Compared with prior art, beneficial effect is the utility model: use the detection of test kit of the present utility model to clinical sample, can in 1.5 hours, complete PCR and detect, sense cycle is short; Each PCR pipe in 96 hole PCR plates can hold different PCR reaction reagents, and a plurality of PCR detection reaction are carried out simultaneously, makes the flux of detection higher; Test kit comprises positive control and negative control, avoids occurring the false positive signal former thereby that occur such as being polluted or due to the false negative signal that reagent inactivation etc. is former thereby occur due to reagent; The PCR testing process of this detection kit adopts fully closed detection, can effectively avoid the environmental pollution of PCR product.
Accompanying drawing explanation
Fig. 1 is the structural representation of the detection kit of the utility model embodiment;
Fig. 2 is the single PCR reaction tubes schematic diagram of the utility model embodiment;
Fig. 3 is negative controls pipe/positive control solution pipe schematic diagram of the utility model embodiment;
Fig. 4 is the sensitivity analysis result schematic diagram that the detection kit of the utility model embodiment is carried out PCR detection.
Embodiment
In order to make the purpose of this utility model, technical scheme and advantage clearer, below in conjunction with drawings and Examples, the utility model is further elaborated.Should be appreciated that specific embodiment described herein is only in order to explain the utility model, and be not used in restriction the utility model.
The utility model embodiment provides a kind of multiple PCR detection kit, comprise box body, lid, supporting seat, negative controls pipe, positive control solution pipe, 96 hole PCR plates, this supporting seat is positioned at this box body top, on this supporting seat, be provided with negative controls pore, positive control solution pore, 96 hole PCR plate holes, this negative controls pore and positive control solution pore are positioned at 96 PCR plate hole the same sides, hole, and this negative controls pipe, positive control solution pipe and 96 hole PCR plates are positioned at respectively this negative controls pore separably, in positive control solution pore and 96 hole PCR plate holes.
Particularly, above-mentioned negative controls pipe is arranged in corresponding negative controls pore separably, and positive control solution pipe is arranged in corresponding positive control solution pore separably, 96 hole PCR plates are arranged in 96 corresponding hole PCR plate holes.Wherein negative controls pore is set to circle, and coordinates with negative controls pipe, and positive control solution pore is also set to circle and coordinates with positive control solution pipe.In specific embodiment, negative controls pipe and positive control solution pipe can be the reagent bottles of same size, also can be the reagent bottle of different size, negative controls pipe and positive control solution pipe can be, but be not limited to one or both in 0.5ml, 1.5ml, 2ml and 5ml screw socket pipe/centrifuge tube.In preferred embodiment, negative controls pipe and positive control solution pipe vary in size to identify and can prevent the mishandles such as application of sample mistake.
Particularly, above-mentioned 96 hole PCR plate holes are rectangular configuration and coordinate so that it is fixed with 96 hole PCR plates.This 96 orifice plate comprises 96 PCR reaction tubess, the structure of this 96 orifice plate can be integrated formed structure, also can be demountable structure, be preferably integrated formed structure so that put into quantitative real time PCR Instrument all detecting PCR reaction tubes simultaneously, in this integrated formed structure, 96 PCR reaction tubess are corresponding with PCR reaction tubes patchhole in quantitative real time PCR Instrument.For demountable structure, this 96 orifice plate is comprised of support and 12 eight unions, can take out in such cases wherein part eight unions and carry out PCR detection and other eight unions are preserved for to later detection.In this 96 orifice plate, for detection of one or more pathogenic agent/microorganism PCR pipes, can represent with different colours or different mark, to avoid wrong application of sample, improve application of sample efficiency.
Fig. 1 has shown a specific embodiment of multiple PCR detection kit structure of the present utility model, wherein this detection kit integral body is rectangular shape, and it comprises box body 1, lid 2, supporting seat 3, negative controls pipe 4, positive control solution pipe 5, amplification toughener pipe 6,96 hole PCR plates 7.Wherein, lid 2 is positioned at box body 1 top, and lid 2 has opening to cover on box body 1 with respect to a side of box body 1.This supporting seat 3 is positioned at box body 1 top, itself and box body 1 are one-body molded, and on this supporting seat 3, are provided with negative controls pore, positive control solution pore, amplification toughener pore and 96 hole PCR plate holes and correspond respectively to negative controls pipe 4, positive control solution pipe 5, amplification toughener pipe 6 and 96 hole PCR plates 7.Negative controls pipe 4, positive control solution pipe 5 and amplification toughener pipe 6 can have identical specification, also can have different specifications, and in Fig. 1, negative controls pipe 4, positive control solution pipe 5 are identical with toughener pipe 6 specifications that increase.In specific embodiment, for ease of distinguishing, prevent wrong application of sample, preferably negative controls pipe 4, positive control solution pipe 5 and amplification toughener pipe 6 are set to different size.The amplification toughener using in the utility model is mainly used in improving pcr amplification product content, can improve PCR detection sensitivity, because amplification toughener consumption is less, therefore preferably the specification of amplification toughener pipe 6 is less than negative controls pipe 4 and positive control solution pipe 5.
Fig. 2 has shown single PCR reaction tubes in the utility model embodiment, and its volume is 200 μ l.In use, eight this kind PCR reaction tubes point-blank can be linked togather, or in the mode of 8x12,96 PCR reaction tubess be linked togather.
Fig. 3 has shown an embodiment of negative controls pipe/positive control solution pipe of the present utility model, and in this embodiment, this negative controls pipe/positive control solution pipe is EP pipe, is 1.5ml specification, is not limited to above-mentioned specification in specific embodiment.
In specific embodiment, above-mentioned negative controls pipe 4, positive control solution pipe 5, amplification toughener pipe 6 sizes and putting in order are not limited to shown in Fig. 1, can adopt other array modes easy to use.
Particularly, in the PCR reaction tubes of above-mentioned negative controls pipe 4, positive control solution pipe 5,96 hole PCR plates 7, corresponding reagent is all housed,, in negative controls pipe 4, negative controls is housed, in positive control solution pipe 5, positive control solution is housed, in the PCR reaction tubes of 96 hole PCR plates 7, PCR reaction solution is housed, this PCR reaction solution comprises PCR damping fluid, MgCl
2, dNTPs, at least one fluorescent probe, at least one pair of PCR primer and archaeal dna polymerase.The DNA sequence dna that contains target detect thing in above-mentioned positive control solution, in negative controls, do not contain this primer sequence, this positive control solution and negative controls the false positive signal former thereby that occur such as are polluted or due to the false negative signal that reagent inactivation etc. is former thereby occur for avoiding occurring due to reagent.Due in above-mentioned PCR reaction tubes, be preinstalled with for detection of PCR reaction solution, therefore after user directly adds testing sample, can detect, shorten like this application of sample flow process, save the time, improve detection efficiency, also avoided user's pollution problem that application of sample may cause repeatedly simultaneously.
Particularly, in each PCR reaction tubes of the multiple PCR detection kit of the utility model embodiment, can comprise the multipair PCR primer for detection of different pathogens,, in each PCR reaction tubes, continue multi-PRC reaction, on the other hand, also can in the different PCR reaction tubess of 96 orifice plates, carry out different PCR reactions, make the flux of detection higher.For example, when detecting for respiratory pathogen, this multiple PCR detection kit can detect 10-12 sample simultaneously, each pattern detection 15-23 kind respiratory pathogen.
Take respiratory pathogen detection below as example, by specific embodiment, the utility model is further set forth.
Embodiment 1PCR specific detection
1. pathogenic micro-organism to be detected comprises as follows: experimental group: respiratory syncytial virus A, metapneumovirus MPV, mycoplasma pneumoniae, legionella; Control group: hepatitis A virus, rotavirus, human cytomegalic inclusion disease virus and hepatitis B virus.
2. the fluorescent PCR of respiratory pathogen detects
(1) clinical samples collection, preservation and transportation
Experimenter is carried out to oropharyngeal swab specimen collection.During collection to be advisable early morning.With clear water, gargle, auxiliary with spatula by examiner, throat swab is crossed to the root of the tongue, arrive the lesion of isthmus faucium portion, repeatedly smear for several times, avoid contacting tongue and oral mucosa etc. and locate during taking-up, the physiological saline of the cotton swab after sampling being put into about 1ml fully washs, then adherent extracting, abandons cotton swab.
(2) sample disposal:
The separation and Culture thing that gathers patient's throat swab extracts nucleic acid from separation and Culture thing.
(3) take patient's nucleic acid carries out RT-PCR reaction as template
In each PCR reaction tubes of 96 hole PCR plates, add nucleic acid samples, in these PCR reaction tubess, be preinstalled with PCR reaction solution.This 96 hole PCR plate is placed in to full-automatic fluorescent PCR detector, and with reference to instrumentation, yin and yang attribute contrast is set in explanation, testing sample parameter is carried out PCR reaction, records sample and puts order.
Wherein, response procedures is set as: the first step: 37 ℃ of 45min, 94 ℃ of 1min; Second step: 94 ℃ of 15sec, 55 ℃ of 15sec, 72 ℃ of 20sec, 40 circulations.
(4) result is judged:
Negative control Ct value is none, and positive control Ct value is less than or equal to 30.It is 35 positive that the Ct value of sample to be tested is less than or equal to, and it is 35 negative that Ct value is greater than.Result shows that test kit specificity is relatively good, to hepatitis A virus, rotavirus, human cytomegalic inclusion disease virus and hepatitis B virus without amplification.Detect respiratory syncytial virus A, metapneumovirus MPV in sample, mycoplasma pneumoniae, a legionella part positive, result does not show.
Above-mentioned testing process can complete in 1.5 hours.
Positive control, by after certain copy number doubling dilution, is detected through pcr amplification until can't detect signal, and this copy number is lowest detectable limit, namely the sensitivity of test kit.Use the maximum sensitivity of test kit of the present utility model can reach 100 copies, see Fig. 4.
The foregoing is only preferred embodiment of the present utility model; not in order to limit the utility model; all any modifications of doing within spirit of the present utility model and principle, be equal to and replace and improvement etc., within all should being included in protection domain of the present utility model.
Claims (8)
1. a multiple PCR detection kit, comprise box body, lid, supporting seat, negative controls pipe, positive control solution pipe, 96 hole PCR plates, described supporting seat is positioned at described box body top, on described supporting seat, be provided with negative controls pore, positive control solution pore, 96 hole PCR plate holes, described negative controls pore and positive control solution pore are positioned at 96 PCR plate hole the same sides, hole, and described negative controls pipe, positive control solution pipe and 96 hole PCR plates are arranged in respectively described negative controls pore, positive control solution pore and 96 hole PCR plate holes separably.
2. multiple PCR detection kit as claimed in claim 1, is characterized in that, the structure of described 96 hole PCR plates be formed in one structure or demountable structure.
3. multiple PCR detection kit as claimed in claim 2, is characterized in that, described demountable structure is comprised of support and 12 eight unions.
4. multiple PCR detection kit as claimed in claim 1, it is characterized in that, also comprise amplification toughener pipe, and on described supporting seat, be provided with amplification toughener pore, described amplification toughener pore and described negative controls pore are positioned at 96 PCR plate hole the same sides, hole, and described amplification toughener pipe is arranged in described amplification toughener pore separably.
5. the multiple PCR detection kit as described in any one in claim 1-4, is characterized in that, described 96 hole PCR plates comprise 96 PCR reaction tubess, in described PCR reaction tubes, PCR reaction solution are all housed.
6. multiple PCR detection kit as claimed in claim 5, is characterized in that, in described negative controls pipe, negative controls is housed, and in positive control solution pipe, positive control solution is housed.
7. multiple PCR detection kit as claimed in claim 5, is characterized in that, described negative controls pipe is different from described positive control solution pipe specification.
8. multiple PCR detection kit as claimed in claim 4, is characterized in that, in described amplification toughener pipe, amplification toughener is housed.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201320686173.4U CN203569116U (en) | 2013-10-31 | 2013-10-31 | Multiple (Polymerase Chain Reaction) detection kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201320686173.4U CN203569116U (en) | 2013-10-31 | 2013-10-31 | Multiple (Polymerase Chain Reaction) detection kit |
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| Publication Number | Publication Date |
|---|---|
| CN203569116U true CN203569116U (en) | 2014-04-30 |
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| CN201320686173.4U Expired - Lifetime CN203569116U (en) | 2013-10-31 | 2013-10-31 | Multiple (Polymerase Chain Reaction) detection kit |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104263640A (en) * | 2014-10-15 | 2015-01-07 | 常州百代生物科技有限公司 | Method for fixing primers or primers and probes on solid-phase carrier for PCR (polymerase chain reaction) |
| CN111647497A (en) * | 2020-05-14 | 2020-09-11 | 南京达伯可特生物科技有限公司 | Multiple closed nucleic acid amplification product rapid detection device |
-
2013
- 2013-10-31 CN CN201320686173.4U patent/CN203569116U/en not_active Expired - Lifetime
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104263640A (en) * | 2014-10-15 | 2015-01-07 | 常州百代生物科技有限公司 | Method for fixing primers or primers and probes on solid-phase carrier for PCR (polymerase chain reaction) |
| CN111647497A (en) * | 2020-05-14 | 2020-09-11 | 南京达伯可特生物科技有限公司 | Multiple closed nucleic acid amplification product rapid detection device |
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| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CX01 | Expiry of patent term |
Granted publication date: 20140430 |
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| CX01 | Expiry of patent term |