CN202193789U - Chip used for typing dengue virus - Google Patents
Chip used for typing dengue virus Download PDFInfo
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- CN202193789U CN202193789U CN2011202570124U CN201120257012U CN202193789U CN 202193789 U CN202193789 U CN 202193789U CN 2011202570124 U CN2011202570124 U CN 2011202570124U CN 201120257012 U CN201120257012 U CN 201120257012U CN 202193789 U CN202193789 U CN 202193789U
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Abstract
The utility model discloses a chip used for typing a dengue virus. The chip used for detecting the serotype of the dengue virus comprises a solid matrix and a probe coating layer arranged on the solid matrix. The chip can detect a plurality of samples simultaneously; therefore, the high throughput detection of the samples is realized, a great quantity of labor cost is saved, and the problem of low throughput in the prior art is solved. Therefore, the chip is of great importance.
Description
Technical field
The utility model relates to a kind of chip that is used for the dengue virus somatotype.
Background technology
Dengue virus (dengue virus) belongs to a serotype subgroup in the flaviviridae Flavivirus, and morphological structure is similar with encephalitis b virus, but volume is less, about 17～25nm.The dengue virus geneome RNA contains 11000 Nucleotide approximately, its 5 ' hold to be I type cap sequence, 3 ' end lacks poly (A) tail, genomic 5 ' end and 3 ' hold one section non-coding region is all arranged.Genome has only an opening code-reading frame, its 5 ' end, 1/4 coding virus 3 structural protein (C, PrM and E), 3 ' end, 3/4 coding 7 Nonstructural Proteins (NS1, NS2a, NS2b, NS3, NS4a, NS4b and NS5).Antigenicity according to virus envelope protein E is different, and dengue virus is divided into 1～4 serotype (I, II, III, IV).Antigenicity has intersection between each C-type virus C, but does not have intercrossing with other antigen crowd of flaviviridae.The antigenic determinant of virus envelope protein E both can induce the host to produce the neutralizing antibody and the hemagglutination inhibition antibody of protectiveness, also possibly participate in the generation of dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS).Detection for dengue virus at present mainly contains viral separation, serodiagnosis and viral nucleic acid detection.Widespread use at present mainly be the RT-PCR method, this method has easy and simple to handle, quick; Susceptibility is high; High specificity; Characteristics low to the parent material specification of quality, but have the high shortcoming of false positive have solved this problem based on the fluorescence quantifying PCR method of TaqMan probe, but still have had the low shortcoming of flux.
The utility model content
The purpose of the utility model provides a kind of chip that is used for the dengue virus somatotype.
The chip that is used to detect dengue virus serotype that the utility model provides comprises that solid substrate and the probe that is arranged on the said solid substrate encapsulate layer.
Wherein, said probe encapsulates the distribution of layer on said solid substrate and is the square formation array.
The column pitch of said square formation array is 10.8mm.
The chip of the utility model can detect a plurality of samples simultaneously, has realized the high throughput testing of sample, has saved a large amount of labour costs, has solved the low problem of prior art flux.Therefore, the utlity model has significance.
The utility model chip can directly be accredited as dengue virus, I type dengue virus, II type dengue virus, III type dengue virus and IV type dengue virus with the dengue virus classification; The experiment proof; This chip has been obtained good effect, and True Positive Rate reaches 100%, and the true negative rate is 100%.Differentiate the rate of accuracy reached to 100% of four type dengue viruss.
Description of drawings
Fig. 1 is the structural representation of chip.Mark is following among the figure: 1 expression solid substrate, 2 expression probes encapsulate layer.
Fig. 2 is the arrangement mode of probe layer on each square formation.
Fig. 3 detects principle for chip uses.
Embodiment
Employed experimental technique is ordinary method like no specified otherwise among the following embodiment.
Used material, reagent etc. like no specified otherwise, all can obtain from commercial sources among the following embodiment.
The structure of embodiment 1, chip
The structure of chip is as shown in Figure 1.
This chip comprises that solid substrate 1 and the probe that is arranged on the solid substrate 1 encapsulate layer 2.And probe encapsulates the distribution of layer 2 on solid substrate 1 and is the square formation array, and the column pitch of square formation array is 10.8mm.
The application of embodiment 2, chip
With the present embodiment is example, and the concrete application method of this chip is described.
One, the kind of probe on the chip
Probe layer comprises as follows: detection probes layer, positive quality control probe layer (PC), negative Quality Control probe layer (NC) and magnetic mark layer;
Said detection probes layer is as follows:
1), be used to detect the general probe of dengue virus, its core hybridization sequences is shown in the SEQ ID NO:5;
2), be used to detect the probe of I type dengue virus, its core hybridization sequences is shown in the SEQ ID NO:1,
3), be used to detect the probe of II type dengue virus, its core hybridization sequences is shown in the SEQ ID NO:2,
4), be used to detect the probe of III type dengue virus, its core hybridization sequences is shown in the SEQ ID NO:3,
5), be used to detect the probe of IV type dengue virus, its core hybridization sequences is shown in the SEQ ID NO:4.
The core hybridization sequences of positive quality control probe is shown in the SEQ ID NO:16
The core hybridization sequences of negative Quality Control probe is shown in the SEQ ID NO:17;
Magnetic target sequence is shown in the SEQ ID NO:18;
Be connected with the straight chain dna fragmentation that is formed by connecting 15 T at 5 of the core hybridization sequences of each probe ' end, and be connected a primary amino with 5 ' end of the core hybridization sequences of each detection probes.
Detection probes, positive quality control probe, negative Quality Control probe and magnetic mark form covalent linkage through the primary amino and the aldehyde radical on the sheet base of its 5 ' end respectively, thereby it is fixed on the sheet base.
The arrangement mode of probe layer is as shown in Figure 2 on each square formation.I is for detecting the probe DEN-1P of dengue virus I type among the figure, and II is for detecting the probe DEN-2P of dengue virus II type, and III is for detecting the probe DEN-3P of dengue virus III type, and IV is for detecting the probe DEN-4P of dengue virus IV type, and T is a general probe.PC is the negative Quality Control probe of hybridization for hybridization positive quality control probe, NC, and B is the magnetic mark.
Solid substrate is the slide sheet base of aldehyde radicalization, available from Boao Biological Co., Ltd (article No. 420022).
Two, preparation method
Be dissolved in the distilled water earlier the probe in the synthetic step 1 well respectively; Obtain various probe solutions; Change the above-mentioned probe solution of 5 μ L respectively in 384 orifice plates or 96 orifice plates; It is (rich difficult to understand biological to add 5 μ L 2 * crystalline substance
gene chip sampling liquid respectively; Catalog number: 440010), and, promptly can be used for gene chip sample applying with behind the pipettor thorough mixing.
Hybridization positive quality control (PC) probe and hybridization negative Quality Control (NC) probe and magnetic mark are dissolved in respectively in the distilled water to final concentration according to also elder generation and are 40 μ M, are used for gene chip sample applying.
With the general and typing probes of above-mentioned detection dengue virus, hybridization positive quality control (PC) probe is set simultaneously, hybridizes negative Quality Control (NC) probe and press point sample step point sample to aldehyde radical sheet base by brilliant core
the micro-array chip point sample system of Boao Biological Co., Ltd's production with magnetic mark (Biotin).
Excellent base of point spends the night at 37 ℃ of water boxes; Washed with de-ionized water 2 times; Chip is put into borate solution (1.0g NaBH4+300ml PBS+100ml absolute ethyl alcohol) hatched 5 minutes; Washed with de-ionized water 2 times behind the air drying, promptly obtains detecting the gene chip of dengue virus general probe and typing probes.The PBS damping fluid of per 1 liter of pH7.4 prepares according to following method: in 1000ml water, add 8.5g NaCl, 2.2g Na
2HPO
4And 0.2g NaH
2PO
4, using HCl or NaOH to transfer PH is 7.4, obtains the PBS damping fluid of pH 7.4.
Chip after preparation is accomplished is through crystalline substance
LuxScan
TMThe 10K micro-array chip scanner scans under the sweep parameter of PMT/Power=90/900, in same chip, and same pin mark appearance, the spot diameter deviation is less than 20%.Dot matrix is neat, does not have obviously to connect a phenomenon, proves that promptly chip is qualified.The result shows that the gene chip of steps A preparation is qualified.
Three, use
(1) reagent that uses is following:
1, in the amplification sample primer of purpose band to compsn, shown in I to V:
Primer is right shown in I, SEQ ID NO:8 and the SEQ ID NO:9; This primer can be hybridized with probe DEN-1 the amplified production of I type dengue virus.
Primer is right shown in II, SEQ ID NO:10 and the SEQ ID NO:11; This primer can be hybridized with probe DEN-2 the amplified production of II type dengue virus.
Primer is right shown in III, SEQ ID NO:12 and the SEQ ID NO:13; This primer can be hybridized with probe DEN-3 the amplified production of III type dengue virus.
Primer is right shown in IV, SEQ ID NO:14 and the SEQ ID NO:15; This primer can be hybridized with probe DEN-4 the amplified production of IV type dengue virus.
Primer is right shown in V, SEQ ID NO:6 and the SEQ ID NO:7; This primer can be hybridized with probe DEN-T the amplified production in dengue virus genome 3 ' non-transcribed zone (UTR).
The right upstream primer 5 ' of above-mentioned primer is held with vitamin H (Biotin) mark.
All added the enzyme recognition site (CTGCAG) of PstI at 5 of forward primer ' end, 5 of reverse primer ' end has added SacI enzyme recognition site (GAGCTC), and has all added the protection base at restriction enzyme site 5 ' end.Add that restriction enzyme site and protection base can improve PCR reaction amplification efficiency.
2, the magnetic bead that encapsulates of Streptavidin becomes biological (112-06D) available from Beijing Si Er.
3, hybridization solution: contain 3 * SSC, 0.2% (quality percentage composition) SDS, 25% (quality percentage composition) methane amide, the solution of 5 * Denhardt ' s.SSC is the damping fluid that contains 0.15M sodium-chlor and 0.015M Trisodium Citrate; Denhardt ' s solution is for containing the mixing solutions of 0.02% (quality percentage composition) ficoll, 0.02% (quality percentage composition) Vinylpyrrolidone polymer and 0.02% (quality percentage composition) BSA.
The number-average molecular weight of ficoll (or weight-average molecular weight) is 4 * 10
5
4, chip scavenging solution:
Chip scavenging solution: washing lotion I: contain 2 * SSC, the aqueous solution of 0.2% (quality percentage composition) SDS; The aqueous solution of washing lotion II:0.2 * SSC.
5, signal reaction damping fluid: the solution that mixes according to 4: 10: 1 volume ratio by sheep blood serum, TEN damping fluid and 1% (quality percentage composition) BSA-PBS solution.Per 1 liter of TEN damping fluid prepares according to following method: Tris-HCl damping fluid, 1m mol EDTA and the 2.0mol NaCl of 10mM, pH7.5 are mixed, supply volume with the Tris-HCl damping fluid of 10mM, pH7.5.Per 1 liter 1% (quality percentage composition) BSA-PBS solution prepares according to following method: 1g BSA is dissolved in the PBS damping fluid of pH 7.4, supplies volume with the PBS damping fluid.PBS damping fluid (pH 7.4) prepares according to following method: in 1000ml water, add 8.5g NaCl, 2.2g Na
2HPO
4And 0.2g NaH
2PO
4, using HCl or NaOH to transfer PH is 7.4, obtains the PBS damping fluid.
6, hybridization positive control sample: its nucleotide sequence is specially shown in the SEQ ID NO:18, and 5 ' end biotin labeling of this dna fragmentation.Use deionized water to set up concentration and be 1ug/ul.
(2) application principle
As shown in Figure 3; Detailed process is with biotin labeled primer sample to be increased; If contain dengue virus in this sample, its corresponding primer dengue virus gene fragment that can increase and obtain biotin labeled corresponding type then, product and chip hybridization that amplification is obtained; The magnetic bead that encapsulates with Streptavidin then carries out mark; If probe can be hybridized combination with amplified production, the marked by magnetic bead that the biotin labeled amplified production that then combines can be encapsulated by Streptavidin, directly visual inspection of reaction result; Or adopt ordinary optical photographic means photographing imaging observations, also can under Power/PMT=90/800, carry out scan image with brilliant core LuxScan 10K/A micro-array chip scanner.Whether contain the dengue virus of this probe in detecting in this sample and can carry out somatotype if the imaging reflecting point is arranged in corresponding position then can confirm it; Otherwise, then do not have.
(3) Application of I
I type dengue virus positive sample: infect I type dengue virus and do not infect the people's of other type dengue viruss serum specimen, take from the Center of Diseases Prevention & Control, Shenzhen City, all pass through patient's agreement; Authentication method is referring to KR Gurukumar; D Priyadarshini, JA Patil, A Bhagat; A Singh.Development of real time PCR for detection and quantitation of Dengue Viruses.Virology Journal; 2009,1,23:1-8).
II type dengue virus positive sample: infect II type dengue virus and do not infect the people's of other types viruses serum specimen, take from the Guangdong Prov. Disease Prevention-control Center, all pass through patient's agreement; Authentication method is referring to KR Gurukumar; D Priyadarshini, JA Patil, A Bhagat; A Singh.Development of real time PCR for detection and quantitation of Dengue Viruses.Virology Journal; 2009,1,23:1-8).
Concrete steps are following:
1, viral RNA extracts and the pcr amplification target gene
1) virus total RNA is extracted
Get 5 parts of I type dengue virus positive sample and 5 parts of II type dengue virus positive sample respectively, get 100ul respectively for every part, add 200ul TRIzol reagent, thermal agitation 60 seconds; Add the 100ul chloroform, thermal agitation 15 seconds, room temperature control 5 minutes, centrifugal 15 minutes then in 12000g; Water is transferred in the new centrifuge tube, added the 500ul Virahol, 4 ℃; Centrifugal 10 minutes of 12000g abandons supernatant, precipitates with 70% washing with alcohol RNA; 12000g is centrifugal 2 minutes then, dry 10 minutes, deposition is dissolved in the DEPC water.
2) reverse transcription
Adopt Omniscript RT kit test kit (the magnificent Sheng Ke in Beijing Bioisystech Co., Ltd; Article No. Qiagen-205111) carries out reverse transcription; Concrete grammar is with random primer 2 μ L, dNTP Mix (10mM) (each 2.5mM) 1 μ L, the total RNA of sample (1ng to 5 μ g) 6 μ L mixings; 65 ℃ of heat denatured 5 minutes, ice bath 5 minutes; Add 10 * First-Strand Buffer, 2 μ L then, MgCL
2(25mM) 4 μ L, DTT (0.1M) 2 μ L, RNaout 1 μ L, mixing, 25 ℃ of heating 2min add SuperScript then
TMII ThermoScript II (5U/uL) 1 μ L, mixing, 25 ℃ of 10min, 42 ℃ of 50min, 70 ℃ of 15min obtain the cDNA of reverse transcription.
3) universal dengue virus gene fragment of pcr amplification and four type dengue virus gene fragments
With above-mentioned steps 2) cDNA that obtains is template, with primer to carrying out pcr amplification reaction.
The PCR reaction system is: 10 * PCR Buffer, 2 μ L, 25mM MgCL
22 μ L; 2.5mM dNTP mix 1.6 μ L; Downstream primer (5 μ M) the 0.5 μ L of downstream primer of dengue virus gene fragment universal amplification primer (5 μ M) and four type dengue virus gene fragment amplification primers; Upstream primer (biotin labeling 40 μ M) the 0.5 μ L of upstream primer of dengue virus gene fragment universal amplification primer (biotin labeling 40 μ M) and four type dengue virus gene fragment amplification primers; The cDNA 2 μ L that the reverse transcription of dengue virus positive sample obtains, LA-TagE (5U/ML) 0.2 μ L, H
2O 11.2 μ L.
The pcr amplification program is: 94 ℃ of 10min of elder generation; 94 ℃ of 30sec then, 51 ℃ of 30sec, 72 ℃ of 30sec, 40 circulations; Last 72 ℃ of 5min.
2, PCR product chip hybridization
According to base complementrity paired principle; Under appropriate reaction conditions; Have biotin labeled PCR product and combine to form stable two strands with complementary probe on the chip, the magnetic bead reaction that encapsulates with Streptavidin again is through the affinity interaction of Streptavidin and vitamin H; After results of hybridization on the chip carried out marked by magnetic bead, obtain the check and analysis result.
Concrete grammar is described below:
1) chip hybridization
Gene chip is put in the hybridizing box; Get the 7.8ul hybridization solution, 0.2ul is hybridized positive control sample, obtains general with 7ul step 2 respectively and the mixing of somatotype dengue virus positive sample PCR product; With the centrifugal 30s of 3000rpm behind the pipettor mixing, 95 ℃ of thermally denature 3min (in the PCR appearance).Ice bath quenching 1min.Annotate on the dot matrix of gene chip through the aperture on the cover plate of hybridizing box with pipettor then, make it cover the dot matrix on the chip.After confirming that hybridization solution covers the dot matrix on the chip, the tight hybridization of lid lid is put into 42 ℃ of thermostat water baths, hybridizes 2 hours, and sample and probe are fully reacted.Annotate: during hybridization bacteria PCR product, need two system PCR products to mix back hybridization.Each dot matrix 15 μ L.Chip totally 4 dot matrix, can hybridize four increments this.
2) chip cleans
The solution of chip cleaning use different ionic strength is the rinsing chip successively, to remove the sample of free and non-specific binding.
With step 2) chip after hybridization finishes; From hybridizing box, take out (cover plate can reuse) fast, chip is transferred in the cleaning box that holds the washing lotion I that is preheated to 42 ℃, hybridization surface up; Be placed on the horizontal shaking table and clean 4min, with flush away not with the sample of probe specific combination.
After in washing lotion I, cleaning end, chip is transferred in the cleaning box that holds the washing lotion II that is preheated to 42 ℃ with tweezers rapidly, hybridization surface is placed on the horizontal shaking table and cleans 4min up.Then, chip was placed in the 50mL taper centrifuge tube 1500rpm centrifugal 1 minute.
3, signal reaction
1) with step 2 each lattice point 12 μ L sheep blood serum of chip after hybridization, hatched 30 minutes for 37 ℃, (prescription is for adding 8.5g NaCl, 2.2g Na respectively in 1000ml water to use 1 * PBS (pH 7.4) then
2HPO
4And 0.2g NaH
2PO
4, using HCl or NaOH to transfer PH is 7.4) and clean centrifugal 1 minute of 1500rpm 2 times.
2) signal reaction
Marked by streptavidin: get the magnetic bead that the Streptavidin after the cleaning encapsulates and mix with 1: 3 volume ratio with the Streptavidin hybridization buffer, put then on the chip that step 1) handled, the point sample amount is the 12ul/ dot matrix, 37 ℃ of hybridization 10 minutes.Use 1 * PBS (to add 8.5g NaCl, 2.2g Na in the 1000ml water respectively then
2HPO
4And 0.2g NaH
2PO
4, using HCl or NaOH to transfer PH is 7.4 solution that obtain) clean 2 times, centrifugal 1 minute of 1500rpm promptly obtains the chip that total overall reaction finishes.
4, interpretation of result
The chip hybridization result can with the naked eye directly distinguish, has reached visual detection requirement directly perceived.The result also can adopt direct optical imaging apparatus to take a picture, like demonstrations such as CCD photo, general camera photos.The result shows that it is positive that the result of 5 parts of I type dengue virus positive sample is I type dengue virus; It is positive that the detected result of 5 parts of II type dengue virus positive sample is II type dengue virus.
(4), Application of I I
According to the method for step (three) to I type dengue virus positive sample (infect I type dengue virus and do not infect the people's of its alloytype dengue virus serum specimen); Detect simultaneously for parallel four times, relatively whether four detected results are investigated chip detecting method and are stablized; The result shows; Detected result all is that positive its alloytype dengue virus of I type dengue virus is negative, between four results and there was no significant difference, proves the chip detecting method quite stable.
(5), Application of I II
With the dna profiling initial amount is that benchmark has carried out preliminary study to the detection sensitivity of this chip.Get I type dengue virus positive, extract viral RNA after reverse transcription, with cDNA carry out successively gradient dilution (100ng, 50ng, 25ng, 10ng), one method is hybridized detection with test kit set by step.CDNA amount detected result shows that the fluorescent signal detection sensitivity is template amount 25ng, and chip detection of the present invention lacks a site when the template amount is reduced to 25ng, and detection sensitivity is template amount 50ng.The result shows that the visual chip detection sensitivity of method of the present invention is cDNA template amount 50ng.
(6), Application of I V
Get clinical detection sample 25 examples (human serum sample is provided by Guangdong Province Disease Control and Prevention Center, all agrees through the patient), detect with test kit; Operate same step 1, detect with PCR method simultaneously that (the PCR detection method is referring to KR Gurukumar, D Priyadarshini; JA Patil, A Bhagat, A Singh.Development of real time PCR for detection and quantitation of Dengue Viruses.Virology Journal; 2009,1,23:1-8).
The result is as shown in table 1 below, and the result shows that the True Positive Rate that detection method of the present invention detects dengue virus is 5/5=100%, and the true negative rate is 20/20=100%, and dengue virus is carried out the rate of accuracy reached to 100% that somatotype detects.
The detected result of table 1. actual sample
| Detection method | PCR detects | Test kit of the present invention |
| The I C-type virus C is positive | ?1 | 1 |
| II type dengue virus is positive | 2 | 2 |
| III type dengue virus is positive | 1 | 1 |
| IV type dengue virus is positive | 1 | 1 |
| Positive sum | 5 | 5 |
| Negative | 20 | 20 |
Sequence table
< 110>Shenzhen Academy of Inspection and Quarantine
< 120>a kind of chip that is used for the dengue virus somatotype
<160>19
<210>1
<211>20
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>1
agagggtgtt?taaagagaaa 20
<210>2
<211>26
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>2
cctaacaatc?ccaccaacag?caggga 26
<210>3
<211>15
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>3
ttggcgaaga?gattc 15
<210>4
<211>15
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>4
atggtgctag?cattc 15
<210>5
<211>27
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>5
agacccccgg?cataacaata?aacagca 27
<210>6
<211>25
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>6
gctgtatcct?ggtggtaagg?actag 25
<210>7
<211>20
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>7
tggtctctcc?cagcgtcaat 20
<210>8
<211>21
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>8
gacaccacac?cctttggaca?a 21
<210>9
<211>17
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>9
tggtgtgcgc?gtgtcaa 17
<210>10
<211>18
<212>DNA
< 213>artificial sequence
<220>
<221>misc_feature
<222>(2)..(2)
< 223>k is g or t
<400>10
tkgtggcgtt?ccttcgtt 18
<210>11
<211>25
<212>DNA
< 213>artificial sequence
<220>
<221>misc_feature
<222>(19)..(19)
< 223>y is t or c
<400>11
tttaattgtt?ccccatctyt?tcagt 25
<210>12
<211>22
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>12
aaccgtgtgt?caactggatc?ac 22
<210>13
<211>20
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>13
gccgttcagc?aatcctcttg 20
<210>14
<211>21
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>14
ttctgggaaa?ggacccttac?g 21
<210>15
<211>20
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>15
ggaaaggact?cgcaaaaacg 20
<210>16
<211>22
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>16
gccgaaccta?ccatacattg?ag 22
<210>17
<211>22
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>17
gccgaaccta?gcatacattg?ag 22
<210>18
<211>171
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>18
agtacccaca?gcacgagaca?cgttcgtcaa?ttcctcgatc?aaggcggctg?ttgattgtgg 60
gtttgacggc?ctggatttac?agtggttgta?cccatcttct?ccgactgata?tgcaaaattt 120
taaaaccctg?attgtgaagt?ggcggcaggc?ggctgtggac?gacgctagaa?a 171
<210>19
<211>15
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>19
tttttttttt ttttt ?15
Claims (3)
1. chip that is used to detect dengue virus serotype is characterized in that: it comprises that solid substrate and the probe that is arranged on the said solid substrate encapsulate layer.
2. chip according to claim 1 is characterized in that: said probe encapsulates the distribution of layer on said solid substrate and is the square formation array.
3. chip according to claim 2 is characterized in that: the column pitch of said square formation array is 10.8mm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011202570124U CN202193789U (en) | 2011-07-20 | 2011-07-20 | Chip used for typing dengue virus |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011202570124U CN202193789U (en) | 2011-07-20 | 2011-07-20 | Chip used for typing dengue virus |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN202193789U true CN202193789U (en) | 2012-04-18 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011202570124U Expired - Fee Related CN202193789U (en) | 2011-07-20 | 2011-07-20 | Chip used for typing dengue virus |
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| Country | Link |
|---|---|
| CN (1) | CN202193789U (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103898240A (en) * | 2014-04-03 | 2014-07-02 | 河北国际旅行卫生保健中心 | Primers, probes and method for detecting dengue virus and types of dengue virus |
| CN104480221A (en) * | 2014-11-21 | 2015-04-01 | 中华人民共和国上海出入境检验检疫局 | Quick high-flux dengue fever virus detection typing method |
-
2011
- 2011-07-20 CN CN2011202570124U patent/CN202193789U/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103898240A (en) * | 2014-04-03 | 2014-07-02 | 河北国际旅行卫生保健中心 | Primers, probes and method for detecting dengue virus and types of dengue virus |
| CN103898240B (en) * | 2014-04-03 | 2016-04-27 | 河北国际旅行卫生保健中心 | Detect the primer of dengue virus and somatotype thereof, probe and method |
| CN104480221A (en) * | 2014-11-21 | 2015-04-01 | 中华人民共和国上海出入境检验检疫局 | Quick high-flux dengue fever virus detection typing method |
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