CN107513494A - A kind of detection of nucleic acids card and its application method - Google Patents
A kind of detection of nucleic acids card and its application method Download PDFInfo
- Publication number
- CN107513494A CN107513494A CN201710705123.9A CN201710705123A CN107513494A CN 107513494 A CN107513494 A CN 107513494A CN 201710705123 A CN201710705123 A CN 201710705123A CN 107513494 A CN107513494 A CN 107513494A
- Authority
- CN
- China
- Prior art keywords
- detection
- nucleic acid
- lines
- card
- well
- 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.)
- Pending
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
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention relates to foranalysis of nucleic acids detection field, spy is related to a kind of detection of nucleic acids card and its application method.Sampling unit of the present invention includes card body, nucleic acid is collected test paper, well and sealed membrane, well and is arranged on card body, and nucleic acid collects test card on well, and well can be sealed membrane closure；Nucleic acid collects test paper and contains cell cracking agent, detection reagent bar is set gradually and formed by sample pad, pad, NC films and adsorptive pads, NC films are C lines, the supporting body of T lines, the flushing that adsorptive pads are used for after nuclease assay reaction, C lines are control line, T lines are detection line, are coated with different nucleic acid specificity antibody on C lines, T lines respectively.The present invention greatly reduces amplification cost, improves amplification efficiency and automaticity, suitable for fields such as experimental study, clinical detection, food safety detections using DNA strict complementary pairing reactions in itself.
Description
Technical field
It is specifically a kind of to be situated between based on nucleic acid amplification reaction and cohesive end the present invention relates to foranalysis of nucleic acids detection field
The nucleic acid detection apparatus and its application method for the strand replacement reaction led.
Background technology
SARS in 2003 breaks out in most of China area, and the epidemic spreads of bird flu in 2005 and 2014 are non-
Ebola's epidemic situation of high fatal rate is wreaked havoc in continent, and a series of this infectious disease as caused by virus is taught that in the development history of the mankind
The mortality that virus is brought is constantly subjected to threaten.The propagation of virus and the variation well beyond mankind's, it is expected that timely and accurately
Diagnosis virus is for improving patient survival, and control epidemic situation is propagated and effectively disease treatment has very great meaning.
Conventional the pathogenic microorganism examination method is to use Vitro Culture Techniques, still, only about 1% bacterium or virus
It can cultivate, and the complexity of incubation and very long incubation time, it greatly limit the popularization of such technology.With point
The development of sub- biology, detection of nucleic acids are applied among Viral diagnosis, are not only shortened detection time and are also improved detection
Sensitivity and specificity.Nucleic acid molecules are genetic carrier of all pathogen including virus, by specific nucleic acid
Detected, it can be determined that with the presence or absence of corresponding virus in sample.PCR is most common detection of nucleic acids side
Method, the appearance of the technology make the medical science detection of nucleic acid level enter a brand-new stage.Round pcr passes through in different temperatures
Between circulation realize the duplication of DNA molecular, the technology is needed to the accurate control of temperature, therefore dependent on accurate temperature control device
Operated with the personnel of specialty.In resource deficient area relative with manpower, PCR method is difficult to use in Viral diagnosis.
In order to overcome the dependence to temperature control device, a series of isothermal amplification methods arise at the historic moment, for example ring mediated isothermal expands
Increase etc..Ring mediated isothermal amplification can expand at 60-65 DEG C to target nucleic acid.Isothermal amplification method can be not only one
Kind quick, sensitive, accurate detection of nucleic acids mode, while simple to temperature control and detection device requirement, significantly reduce equipment into
This input., can be to sample using the cooperation optical detection of DNA fluorescence indicators or the Turbidity measurement based on magnesium pyrophosphate accessory substance
Product carry out quantitative analysis.
After the strand replacement reaction of cohesive end mediation refers to the single-stranded hybridization of DNA of two complete complementaries or partial complementarity, put
The process of the DNA of one or more prehybridization is changed, the reaction is a dynamic competition process.This reaction starts from target
DNA pendency end is cohesive end.What TSD was relied on is DNA strict base pair complementarity properties in itself, it is not necessary to nuclease
Participation can occur.Therefore under conditions of the direct anneal of DNA double chain, the reaction can pass through the single-stranded pendency of complementation
Region is that cohesive end triggers, and then undergoes a branch migration process and completes.
In clinical case, same illness may be produced by a variety of different pathogen, cause diagnosis with suiting the medicine to the illness
The difficulty write a prescription.Using nucleic acid amplification method, either PCR or isothermal duplication, how to detect simultaneously multiple pathogens or
Target gene is always a significant challenge in clinical detection.Therefore, invention one kind is simple to operate, the degree of accuracy is high, sensitivity
High detection of nucleic acids product, is detected significant while realizing multiple pathogens or target gene.
At present, precisely medical treatment has turned into the study hotspot and development trend of global modern medical service, in the world relevant precisely doctor
The research and achievements conversion for the treatment of surge forward.Plan proposition and greatly develop accurate medical treatment in China 13 so that in this science and technology
Field China there will be an opportunity to realize that bend is overtaken other vehicles, and walk the rank of advanced units in world's medical science technology.Precisely medical treatment includes accurate
Diagnosis and precisely treatment, wherein diagnosis is the basis for the treatment of, and the diagnosis of the molecule based on nucleic acid detection technique is then precisely to diagnose
Core.China or even in the world current molecule diagnose market and are based primarily upon various large-scale instrument and equipments and tediously long complicated tissue
Sample pretreatment means, have the shortcomings that costly, time-consuming, target is inaccurate, popularization is not high.The present invention is to be based on miniflow
Control chip technology novel molecular diagnosing chip, than prior-generation technology have high flux, low cost, rapidly and efficiently, sensitivity
The advantages that high, with strong points, universality is strong, there is very wide application potential and market prospects.Compared with prior art can only
Take the mode of " sweeping entirely " that the full-length genome of diagnosis and treatment object is sequenced, cause ample resources waste and data it is meaningless
Repeat so that the present situation that Consumer groups hang back to high sequencing expense.
The content of the invention
In view of the shortcomings of the prior art, the strand displacement that the present invention is mediated by nucleic acid amplification reaction or cohesive end is anti-
Should, not only can visually carry out qualitative analysis also can carry out quantitative analysis by optical detector, realize to single sample or
Multiple pathogens or target gene in multisample detect simultaneously, detect and take short, the degree of accuracy and high sensitivity, and profit
Nucleic acid amplification enzyme and triphosphate deoxy-nucleotide usually required in nucleic acid amplification reaction are avoided with TSDR reaction principles, is utilized
DNA strict complementary pairing reactions in itself, greatly reduce amplification cost, improve amplification efficiency and automaticity, be applicable
In fields such as experimental study, clinical detection, food safety detections.
The technical scheme is that：A kind of detection of nucleic acids card, including sampling unit, detection unit, it is characterised in that：Institute
The sampling unit stated includes card body, nucleic acid is collected test paper, well and sealed membrane, well and is arranged on card body, and nucleic acid is collected
For test card on well, well can be sealed membrane closure；Described nucleic acid collects test paper and contains cell cracking agent, described
Detection unit include detection reagent bar, described detection reagent bar is set gradually by sample pad, pad, NC films and adsorptive pads
Composition, pad are the carriers of the specific nucleic acid identification fragment of nanogold particle mark, and sample pad is to be used to carry micro sample
The carrier for the amplified production that this strand replacement reaction mediated through nucleic acid amplification reaction or cohesive end obtains, NC films are C lines, T
The supporting body of line, the flushing that adsorptive pads are used for after nuclease assay reaction, C lines are control line, and T lines are detection line, are divided on C lines, T lines
Different nucleic acid specificity antibody is not coated with.
According to detection of nucleic acids card as described above, it is characterised in that：Described detection unit also includes upper shell, lower house
And insulating cushion, upper shell are provided with detection window, lower house is anterior to include the outpost of the tax office one, the outpost of the tax office two, insulating cushion neck and water suction paper card
Groove, insulating cushion are placed in insulating cushion neck, and blotting paper is placed in blotting paper neck, and the outpost of the tax office is set respectively on the inside of lower house
First, the outpost of the tax office two, the rear portion of lower house is bogey, and detection reagent bar is arranged on bogey, and upper shell is connected to lower casing
After body, detection window face C lines, T lines.
According to detection of nucleic acids card as described above, it is characterised in that：Described card body material is plastics.
According to detection of nucleic acids card as described above, it is characterised in that：Described sealing membrane material is plastics, and sealed membrane passes through
Viscose glue or electrostatic interaction are fixed on card body.
According to detection of nucleic acids card as described above, it is characterised in that：Described pad and sample pad material are glass fibers
Dimension.
According to detection of nucleic acids card as described above, it is characterised in that：Described NC membrane materials are nitrocellulose filter.
According to detection of nucleic acids card as described above, it is characterised in that：Also include reaction reagent, described reaction reagent is core
Sour amplifing reagent, nucleic acid amplification agents include the enzyme needed for primer, nucleic acid amplification reaction, buffer solution, triphosphate deoxy-nucleotide,
The strand replacement reaction reagent of cohesive end mediation includes primer, fluorogen, biotin.
According to detection of nucleic acids card as described above, it is characterised in that：Also include buffer solution, described buffer solution delays for phosphoric acid
Fliud flushing.
According to detection of nucleic acids card as described above, it is characterised in that：Also include detector, described detector is adopted by optics
Collect unit, image-signal processor and electronics peripheral hardware composition, detection of nucleic acids result is read by optical detection, it is included in liquid
On brilliant screen, and print.
The invention also discloses a kind of application method of detection of nucleic acids card, comprise the following steps：
Detection reagent bar is placed on the bogey at lower house rear portion, insulating cushion is placed on insulating cushion neck,
Blotting paper is placed on blotting paper neck, upper shell and lower house are connected together, and card body is enclosed on upper shell and lower house
Front portion, well is in directly over blotting paper neck；
Test paper is collected to nucleic acid by well and is respectively dropped into sample and buffer solution, is dried after standing, card body is pushed away forward
To the outpost of the tax office two, well is set to be in directly over insulating cushion neck；
Reaction solution is instilled into well, reaction solution collects test paper with nucleic acid and insulating cushion contacts, then using sealed membrane
Well is closed, and is positioned in baking oven and reacts；
Reaction is finished, and card body is pushed forward into the outpost of the tax office one, well is in directly over sample pad, removes sealed membrane, to
Buffer solution is instilled in well, then observes the change of C line, T lines.
The beneficial effects of the invention are as follows：Using a detection means, multiple nucleic acid amplification reaction systems are pre-stored in nucleic acid
In detection card, by nucleic acid amplification reaction or the strand replacement reaction of cohesive end mediation, using chromatographing, effect, nucleic acid amplification are anti-
The strand replacement reaction that should be mediated with cohesive end, not only can visually carry out qualitative analysis can also be determined by optical detector
Amount analysis, realize and the multiple pathogens in single sample either multisample or target gene are carried out while detected, detection consumption
When short, the degree of accuracy and high sensitivity, and avoid in nucleic acid amplification reaction usually required nucleic acid using TSDR reaction principles and expand
Increase enzyme and triphosphate deoxy-nucleotide, strict complementary pairing reacts in itself using DNA, greatly reduces amplification cost, improves
Amplification efficiency and automaticity, suitable for fields such as experimental study, clinical detection, food safety detections.
Brief description of the drawings
Fig. 1 is upper shell structural representation of the present invention；
Fig. 2 is lower house structural representation of the present invention；
Fig. 3 is detection reagent bar structural representation；
Fig. 4 is insulating cushion schematic diagram；
Fig. 5 is sampling unit schematic diagram；
Fig. 6 is sealed membrane schematic diagram；
Fig. 7 is blotting paper schematic diagram.
Description of reference numerals：Upper shell 1, detection window 2, lower house 3, the outpost of the tax office 1, the outpost of the tax office 25, insulating cushion neck 6, water suction
Paper card groove 7, nucleic acid collect test paper 8, card body 9, well 10, adsorptive pads 11, C lines 12, T lines 13, NC films 14, pad 15, sample
Product pad 16, insulating cushion 17, blotting paper 18, sealed membrane 19.
Embodiment
Technical scheme is described in further detail below in conjunction with accompanying drawing.
As shown in figure 1, the detection of nucleic acids card of the present invention includes sampling unit, detection unit, in test process, in addition to
Matching used reaction reagent, buffer solution and detector.
As shown in figure 5, sampling unit includes card body 9, nucleic acid collects test paper 8, well 10 and sealed membrane 19, well 10
It is arranged on card body 9, nucleic acid is collected test paper 8 and is stuck on well 10, and well 10 can be sealed film 19 and close.In detection process
In, trace sample, reaction solution, buffer solution add by well 10, for residue and unnecessary buffer solution, can pass through water suction
Paper 18 is drawn, and after the completion of sample-adding, can be sealed well 10 using sealed membrane 19.The material of card body 9 of the present invention is plastics, in core
In sour detection process, card body 9 can be mobile to detection unit direction and positioned by the outpost of the tax office 1 in detection unit or the outpost of the tax office 25, is used
To complete the strand replacement reaction of the nucleic acid amplification reaction of detection of nucleic acids process or cohesive end mediation and nucleic acid specificity/non-spy
The opposite sex combines process color.
Amplifying nucleic acid of the present invention collects test paper 8 and contains cell cracking agent, can make protein denaturation and effective with cell lysis
Protect nucleic acid from nuclease, oxidant and ultraviolet injury in ground.It is anti-nucleic acid amplification can to occur on nucleic acid collection test paper 8 of the present invention
Should, nucleic acid amplification reaction of the invention can be anti-as polymerase chain reaction, the ring mediated isothermal amplification of amplification template using DNA
, rolling circle amplification reaction, the isothermal amplification of unwindase, the isothermal amplification of recombinase-mediated or other nucleic acid should be relied on
Amplified reaction or RT-polymerase chain reaction, reverse transcription loop-mediated isothermal amplified reaction using RNA as amplification template
Or other reverse transcription nucleic acid amplification reactions.
The material of sealed membrane 19 of the present invention can be plastics, and sealed membrane 19 is fixed on card body 9 by viscose glue or electrostatic interaction,
Realize the sealing to well 10.
As shown in Figures 1 to 4, detection unit includes upper shell 1, lower house 3, detection reagent bar with upper shell clamping
With insulating cushion 17, upper shell 1 is provided with detection window 2, and lower house 3 is anterior to include the outpost of the tax office 1, the outpost of the tax office 25, the and of insulating cushion neck 6
Blotting paper neck 7, insulating cushion 17 are placed in insulating cushion neck 6, and blotting paper 18 is placed in blotting paper neck 7, under
The inner side of housing 3 sets the outpost of the tax office 1, the outpost of the tax office 25 respectively, and the outpost of the tax office 1, the outpost of the tax office 25 are used to position card body 9.Lower house 3
Rear portion is bogey, for carrying detection reagent bar, after upper shell 1 is connected to lower house 3, and detection window 2 face C lines 12, T lines
13, so it is easy to directly observation or apparatus measures test result.
As shown in figure 3, the present invention detection reagent bar by sample pad 16, pad 15, NC films 14 and adsorptive pads 11 successively
Composition is set, and pad 15 is the carrier of the specific nucleic acid identification fragment of nanogold particle mark, and sample pad 16 is to be used to hold
Carry the carrier for the amplified production that the strand replacement reaction that trace sample mediates through nucleic acid amplification reaction or cohesive end obtains, NC films
14 be C lines 12, the supporting body of T lines 13, and the flushing that adsorptive pads 11 are used for after nuclease assay reaction, C lines 12 are control line, T lines
13 be detection line, and in the presence of only C lines 12, testing result is feminine gender, when C lines 12, T lines 13 then detect knot in the presence of simultaneously
Fruit is the positive.
The material of sample pad 16 of the present invention is glass fibre, for carrying trace sample through nucleic acid amplification reaction or viscosity
The amplified production that the strand replacement reaction of end mediation obtains, and it is directed to pad 15 and NC films 14, and and NC through adsorptive pads 11
Coated C lines 12 on film 14, coated specificity/non-specific identification nucleic acid fragment reaction on T lines 13；
The material of pad 15 is glass fibre, and fragment is identified for carrying the specific nucleic acid through nanogold particle mark,
To capture the sample nucleic acid fragment amplification product in sample pad 16；
The material of NC films 14 is nitrocellulose filter, C lines 12, the T lines 13 used for carrying detection of nucleic acids colour developing, C lines
12nd, different nucleic acid specificity antibody is coated with T lines 13 respectively, for combining the specific recognition nucleic acid of nanogold particle mark
Fragment and the compound of sample nucleic acid amplified production hybridization.
The matching used reaction reagent of the present invention can be nucleic acid amplification agents, and nucleic acid amplification agents include primer, nucleic acid
Enzyme, buffer solution, triphosphate deoxy-nucleotide needed for amplified reaction, the strand replacement reaction reagent of cohesive end mediation include drawing
Thing, fluorogen, biotin, not including the enzyme and triphosphate deoxy-nucleotide needed for nucleic acid amplification, be pre-installed on detection of nucleic acids card with
In the containers such as outer test tube, in case using.
The buffer solution of the present invention can use phosphate buffer etc., and buffer solution is pre-installed on test tube beyond detection of nucleic acids card etc.
In container, in case using.
The present invention the matching used detector of nucleic acid detection apparatus, by optically detecting unit, image-signal processor and
Associated electrical peripheral hardware forms, and reads detection of nucleic acids result by optical detection, it is included on LCD screen, and prints
Below by a specific embodiment, the present invention is described further, but specific embodiment is not to this
Any restriction is done in invention.
Example 1：The site primer of epithelial growth factor receptor gene 19 in lung ED-SCLC
Nucleic acid detection apparatus square is taken out on experimental bench, 1 drop sample is added into well 10, is stored at room temperature 5 minutes；
1) continue to add 1-2 drop buffer solutions into well 10, after standing 5 minutes, card body 9 is pushed forward to the outpost of the tax office 25
Position；
2) add 1 drop reaction solution into well 10, while sealed membrane 19 is closed into well 10, detection means is placed in 65
1 hour or so in DEG C baking oven；
Take out detection means, remove sealed membrane 19, is continued to push forward at the outpost of the tax office 1, be added dropwise 1 drip buffer solution in
In well 10, the change of C lines 12, T lines 13 is visually can be seen that in 5 minutes.In the presence of only C lines 12, testing result is
Feminine gender, when C lines 12, T lines 13 simultaneously in the presence of then testing result for the positive, can also be put into the matching used inspection of nucleic acid detection apparatus
Survey in device and quantitatively detect corresponding result.
The invention also discloses the application method of detection of nucleic acids card, comprise the following steps：
Detection reagent bar is placed on the bogey at the rear portion of lower house 3, insulating cushion is placed on insulating cushion neck 6
17, blotting paper 18 is placed on blotting paper neck 7, upper shell 1 and lower house 3 are connected together, and card body 9 is enclosed on
The front portion of housing 1 and lower house 3, well 10 is set to be in directly over blotting paper neck 7；
By well 10 to nucleic acid collect test paper 8 be respectively dropped into sample and buffer solution, dried after standing, by card body 9 to
Before shift the outpost of the tax office 25 onto, well 10 is in directly over insulating cushion neck 6；
Then reaction solution is instilled into well 10, reaction solution collects test paper 8 with nucleic acid and insulating cushion 17 contacts, then
Well 10 is closed using sealed membrane 19, and is positioned in baking oven and reacts；
Reaction is finished, and card body 9 is pushed forward into the outpost of the tax office 1, well 10 is in directly over sample pad 16, removes envelope
Membrana oralis 19, buffer solution is instilled into well 10, then observe C lines 12, the change of T lines 13.
The micro-fluidic diagnosing chip technology of high flux that the present invention is taken then has abandoned side used by background technology completely
Formula mode, but pointedly target gene fragment is expanded with the reaction system of " section type " or signal amplifies and carried out
The ultrasensitiveness detection of direct-reading, it is achieved thereby that being complemented each other with the tentative diagnosis of clinician and being accurate clinical diagnosis
Strong reliable foundation is provided, precisely diagnosis is really become a reality with precisely medical treatment.And in emerging pathogen detection field,
The micro-fluidic molecular diagnosis chip technology of high flux high sensitivity that the present invention develops is even more to have conventional molecular diagnostic techniques institute not
Analogous technical advantage.Due to rapid amplifying of this technology based on the extremely strong pathogenic microorganism genetic fragment of purpose rather than
" scanning " to pathogenic microorganism full-length genome, so the molecular diagnosis chip quickly can accurately judge Pathogen category sum
Amount.This achievement by cause China at present still part there is abuse of antibiotics problem thoroughly turn into history, it is market-oriented
There to be immeasurable Social benefit and economic benefit.
The market orientation of the present invention can tentatively be set to：1. pathogen detection.The development side of pathogen detection technology at present
Xing Weiai is still serious the problem of abuse of antibiotics.The present invention can make it that the specific aim of clinical application is more accurate, high degree
Avoid the economic waste of mistaken diagnosis and patient in medication.2. disease gene detects.Different from carrying out full genome survey to human body
The mode of sequence, using the micro-fluidic molecular diagnosis chip technology based on high flux high sensitivity, clinical gene diagnosis and gene wind
Danger diagnosis will enter into huge numbers of families, and miniaturization, rapid, economical and practical type portable gene diagnosis instrument will cause doctor and trouble
Person more understands to its gene risk and disease risk, realizes and the specific aim of disease is accurately prevented and treated.
As it will be easily appreciated by one skilled in the art that the foregoing is merely illustrative of the preferred embodiments of the present invention, not to
The limitation present invention, all any modification, equivalent and improvement made within the spirit and principles of the invention etc., all should be included
Within protection scope of the present invention.
Claims (10)
1. a kind of detection of nucleic acids card, including sampling unit, detection unit, it is characterised in that：Described sampling unit include card body,
Nucleic acid is collected test paper, well and sealed membrane, well and is arranged on card body, and nucleic acid collects test card on well, sample-adding
Hole can be sealed membrane closure；Described nucleic acid collects test paper and contains cell cracking agent, and described detection unit includes detection and tried
Agent bar, described detection reagent bar are set gradually and formed by sample pad, pad, NC films and adsorptive pads, and pad is nanogold
The carrier of the specific nucleic acid identification fragment of particle marker, sample pad be used to carry trace sample through nucleic acid amplification reaction or
The carrier for the amplified production that the strand replacement reaction of cohesive end mediation obtains, NC films are C lines, the supporting body of T lines, and adsorptive pads are used for
Flushing after nuclease assay reaction, C lines are control line, and T lines are detection line, and different nucleic acid specificities is coated with respectively on C lines, T lines
Property antibody.
2. detection of nucleic acids card according to claim 1, it is characterised in that：Described detection unit also includes upper shell, under
Housing and insulating cushion, upper shell are provided with detection window, and lower house front portion includes the outpost of the tax office one, the outpost of the tax office two, insulating cushion neck and water suction
Paper card groove, insulating cushion are placed in insulating cushion neck, and blotting paper is placed in blotting paper neck, are set and are closed respectively on the inside of lower house
Card one, the outpost of the tax office two, the rear portion of lower house is bogey, and detection reagent bar is arranged on bogey, and upper shell is connected to down
After housing, detection window face C lines, T lines.
3. detection of nucleic acids card according to claim 1 or 2, it is characterised in that：Described card body material is plastics.
4. detection of nucleic acids card according to claim 1 or 2, it is characterised in that：Described sealing membrane material is plastics, sealing
Film is fixed on card body by viscose glue or electrostatic interaction.
5. detection of nucleic acids card according to claim 1 or 2, it is characterised in that：Described pad and sample pad material be
Glass fibre.
6. detection of nucleic acids card according to claim 1 or 2, it is characterised in that：Described NC membrane materials are nitrocellulose
Film.
7. detection of nucleic acids card according to claim 1 or 2, it is characterised in that：Also include reaction reagent, described reaction examination
Agent is nucleic acid amplification agents, and nucleic acid amplification agents include the enzyme needed for primer, nucleic acid amplification reaction, buffer solution, triphosphoric acid deoxidation
Nucleotides, the strand replacement reaction reagent of cohesive end mediation include primer, fluorogen, biotin.
8. detection of nucleic acids card according to claim 1 or 2, it is characterised in that：Also include buffer solution, described buffer solution is
Phosphate buffer.
9. detection of nucleic acids card according to claim 1 or 2, it is characterised in that：Also include detector, described detector by
Optically detecting unit, image-signal processor and electronics peripheral hardware composition, detection of nucleic acids result is read by optical detection.
10. a kind of application method of detection of nucleic acids card, comprises the following steps：
Detection reagent bar is placed on the bogey at lower house rear portion, insulating cushion is placed on insulating cushion neck, absorbed water
Blotting paper is placed on paper card groove, upper shell and lower house are connected together, and before card body is enclosed on into upper shell and lower house
Portion, well is set to be in directly over blotting paper neck；
Test paper is collected to nucleic acid by well and is respectively dropped into sample and buffer solution, is dried after standing, card body is pushed forward to pass
Card two, well is set to be in directly over insulating cushion neck；
Reaction solution is instilled into well, reaction solution collects test paper with nucleic acid and insulating cushion contacts, then using sealing membrane closure
Well, and be positioned in baking oven and react；
Reaction is finished, and card body is pushed forward into the outpost of the tax office one, well is in directly over sample pad, sealed membrane is removed, to sample-adding
Buffer solution is instilled in hole, then observes the change of C line, T lines.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710705123.9A CN107513494A (en) | 2017-08-17 | 2017-08-17 | A kind of detection of nucleic acids card and its application method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710705123.9A CN107513494A (en) | 2017-08-17 | 2017-08-17 | A kind of detection of nucleic acids card and its application method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107513494A true CN107513494A (en) | 2017-12-26 |
Family
ID=60723493
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710705123.9A Pending CN107513494A (en) | 2017-08-17 | 2017-08-17 | A kind of detection of nucleic acids card and its application method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107513494A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109198732A (en) * | 2018-08-20 | 2019-01-15 | 苏州晞光生物科技有限公司 | A kind of clothes and test method of the wearable detection human secretion of intelligence |
| CN110218769A (en) * | 2019-07-08 | 2019-09-10 | 中国人民解放军东部战区疾病预防控制中心 | The multi-joint test strips of integration integral type for LAMP or RT-LAMP |
| CN116606727A (en) * | 2023-04-17 | 2023-08-18 | 长庚大学 | Nucleic acid detection device and nucleic acid detection method |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060099596A1 (en) * | 2002-12-06 | 2006-05-11 | Roche Molecular Systems, Inc. | Multiplex assay detection of pathogenic organisms |
| US20110039261A1 (en) * | 2007-12-20 | 2011-02-17 | Aj Innuscreen Gmbh | Mobile rapid test system for nucleic acid analysis |
| CN201788183U (en) * | 2010-03-02 | 2011-04-06 | 珠海市银科医学工程有限公司 | Bacterial vaginosis joint detecting card |
| CN104278079A (en) * | 2013-07-08 | 2015-01-14 | 嘉兴朝云帆生物科技有限公司 | Test strip and method for detecting nucleic acid through nucleic acid chromatographic technique |
| CN105273997A (en) * | 2015-10-21 | 2016-01-27 | 西安交通大学 | Paper-based nucleic acid extraction, amplification and detection integrated detection system and paper-based nucleic acid extraction, amplification and detection integrated detection method |
| CN205024226U (en) * | 2015-04-30 | 2016-02-10 | 美利泰格诊断试剂(嘉兴)有限公司 | Test panel is surveyed to pair spirillum |
| CN106337085A (en) * | 2016-08-24 | 2017-01-18 | 冯晓均 | Nucleic acid test card and application method thereof |
| CN106636387A (en) * | 2016-12-14 | 2017-05-10 | 天津科技大学 | Salmonella nucleic acid rapid detection kit, test strip and detection method |
| CN207512174U (en) * | 2017-08-17 | 2018-06-19 | 湖北伽诺美生物科技有限公司 | A kind of detection of nucleic acids card |
-
2017
- 2017-08-17 CN CN201710705123.9A patent/CN107513494A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060099596A1 (en) * | 2002-12-06 | 2006-05-11 | Roche Molecular Systems, Inc. | Multiplex assay detection of pathogenic organisms |
| US20110039261A1 (en) * | 2007-12-20 | 2011-02-17 | Aj Innuscreen Gmbh | Mobile rapid test system for nucleic acid analysis |
| CN201788183U (en) * | 2010-03-02 | 2011-04-06 | 珠海市银科医学工程有限公司 | Bacterial vaginosis joint detecting card |
| CN104278079A (en) * | 2013-07-08 | 2015-01-14 | 嘉兴朝云帆生物科技有限公司 | Test strip and method for detecting nucleic acid through nucleic acid chromatographic technique |
| CN205024226U (en) * | 2015-04-30 | 2016-02-10 | 美利泰格诊断试剂(嘉兴)有限公司 | Test panel is surveyed to pair spirillum |
| CN105273997A (en) * | 2015-10-21 | 2016-01-27 | 西安交通大学 | Paper-based nucleic acid extraction, amplification and detection integrated detection system and paper-based nucleic acid extraction, amplification and detection integrated detection method |
| CN106337085A (en) * | 2016-08-24 | 2017-01-18 | 冯晓均 | Nucleic acid test card and application method thereof |
| CN106636387A (en) * | 2016-12-14 | 2017-05-10 | 天津科技大学 | Salmonella nucleic acid rapid detection kit, test strip and detection method |
| CN207512174U (en) * | 2017-08-17 | 2018-06-19 | 湖北伽诺美生物科技有限公司 | A kind of detection of nucleic acids card |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109198732A (en) * | 2018-08-20 | 2019-01-15 | 苏州晞光生物科技有限公司 | A kind of clothes and test method of the wearable detection human secretion of intelligence |
| CN110218769A (en) * | 2019-07-08 | 2019-09-10 | 中国人民解放军东部战区疾病预防控制中心 | The multi-joint test strips of integration integral type for LAMP or RT-LAMP |
| CN110218769B (en) * | 2019-07-08 | 2024-04-19 | 中国人民解放军东部战区疾病预防控制中心 | Integrated multi-linked test strip for LAMP or RT-LAMP |
| CN116606727A (en) * | 2023-04-17 | 2023-08-18 | 长庚大学 | Nucleic acid detection device and nucleic acid detection method |
| CN116606727B (en) * | 2023-04-17 | 2024-03-12 | 长庚大学 | Nucleic acid detection device and nucleic acid detection method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102154498B (en) | Nucleic acid detecting method | |
| CN104342503B (en) | A kind of method simultaneously detecting 12 kinds of common Respiroviruses | |
| CN105392896B (en) | Quick and sensitive genotype identification and detection of nucleic acids | |
| CN107475446B (en) | Multiplex PCR detection method for simultaneously detecting multiple respiratory viruses, probe set and kit thereof | |
| CN103074448B (en) | Kit for synchronously detecting twenty-three meningitis pathogens and detection method of kit | |
| CN105400907A (en) | Kit for nucleic acid combined detection of influenza virus A, influenza virus B and respiratory syncytial virus | |
| Bearinger et al. | Development and initial results of a low cost, disposable, point-of-care testing device for pathogen detection | |
| CN103074451A (en) | Kit for synchronously detecting twenty-two respiratory tract pathogens and detection method of kit | |
| CN103074434A (en) | CYP2C19 gene polymorphyism detection kit and detection method thereof | |
| CN108588277A (en) | A kind of canine distemper virus visualization nucleic acid detection method | |
| CN107513494A (en) | A kind of detection of nucleic acids card and its application method | |
| CN106434996A (en) | Kit and method for detecting Acinetobacter baumannii DNA | |
| CN103074452A (en) | Kit for synchronously detecting fifteen hemorrhagic fever pathogens and detection method of kit | |
| CN106337085A (en) | Nucleic acid test card and application method thereof | |
| CN104561372A (en) | Combined primer for amplification and typing of human papilloma virogenes and application of combined primer | |
| CN106148484B (en) | A kind of kit that diagnosis Y chromosome is micro-deleted | |
| Gou et al. | The colorimetric isothermal multiple-self-matching-initiated amplification using cresol red for rapid and sensitive detection of porcine circovirus 3 | |
| CN105803062A (en) | Nucleic acid (DNA/RNA) real-time constant-temperature gene amplification detection method | |
| ITVT20110002A1 (en) | METHOD OF DETERMINING THE ORIGIN OF FLUIDS OR BIOLOGICAL TRACKS AND REAGENT KITS FOR THEIR IDENTIFICATION IN A SAMPLE. | |
| CN105734158A (en) | Fluorescent PCR (polymerase chain reaction) detection kit for babesia caballi disease | |
| CN109355406B (en) | A kind of kit of the detection mycobacterium tuberculosis based on blood free nucleic acid | |
| CN108531627A (en) | One kind is for detecting the streptococcic RPA fluorescent quantitations primer pair of B races, probe, kit and detection method | |
| CN108048589A (en) | The Real-time PCR specific primers and probe and detection kit and detection method of ox two-pressure humidity generator | |
| Li et al. | Visual and rapid detection of porcine epidemic diarrhea virus (PEDV) using reverse transcription loop-mediated isothermal amplification method | |
| CN101591713B (en) | Gosling plague virus LAMP detection kit and detection method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| AD01 | Patent right deemed abandoned |
Effective date of abandoning: 20221230 |
|
| AD01 | Patent right deemed abandoned |