CN105385766B - A kind of Streptococcus suis Serotypes liquid phase chip reagent box - Google Patents
A kind of Streptococcus suis Serotypes liquid phase chip reagent box Download PDFInfo
- Publication number
- CN105385766B CN105385766B CN201510954076.2A CN201510954076A CN105385766B CN 105385766 B CN105385766 B CN 105385766B CN 201510954076 A CN201510954076 A CN 201510954076A CN 105385766 B CN105385766 B CN 105385766B
- Authority
- CN
- China
- Prior art keywords
- seq
- specific
- phase chip
- streptococcus suis
- specificity
- 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.)
- Active
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
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
-
- 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
-
- 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
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/16—Primer sets for multiplex assays
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (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
A kind of Streptococcus suis Serotypes liquid phase chip reagent box of the present invention, the upstream primer probe sequence of the upstream primer probe, specificity that contain specificity is as shown in NO:41 ~ 60 SEQ ID；The sequence of the downstream primer probe of downstream primer probe and specificity also containing specificity is that added one section of biotin at 5 ' ends of any one of specific downstream primer probe shown in SEQ ID NO:2,4,6,8,10,12,14,16,18,20,22,24,26,28,30,32,34,36,38,40；Also containing specific microballoon, the sequence of specific microballoon is as shown in NO:61 ~ 80 SEQ ID.The present invention, which is realized, can carry out the detection of liquid-phase chip parting to 20 boar streptococcus serum types simultaneously in a reaction system, have the characteristics that high speed, high-throughput, high specific and accuracy are high.
Description
Technical field
The invention belongs to field of biological genes, are related to a kind of Serotypes liquid phase chip reagent box of bacterium, specific next
Say it is a kind of Streptococcus suis Serotypes liquid phase chip reagent box.
Background technique
Nineteen fifty-one, Holland reported Streptococcus suis type earliest, hereafter had generation in the other countries of aquaculture prosperity.Pig
Streptococcosis just has generation early in 20th century 50, the sixties, falls ill and increases after the seventies, the Xianyang region of Sichuan in 2005 is broken out
Streptococcus suis 2-type epidemic situation, and the infection and death of related personnel are caused, so that Streptococcus suis has been a great concern.
Currently, the method for Streptococcus suis Serotypes mainly has serum agglutination or coagglutination, but the method is deposited
In the shortcomings that time-consuming, as a result hardly possible judges, in screening a large amount of clinical samples, workload is very big, and introduces a set of pig hammer
Bacterium blood grouping serum price wants more than 20 ten thousand, and is only sufficient to identification 50 times.Molecules context of detection is also only directed to serotype 1 so far
(14), 2 (1/2), 7,9 types PCR detection method, with the complication of epidemic situation and further expanding for research contents, only this 6
The detection method of a serotype has been unable to meet needs.On current veterinary clinic and public health prevention and control, urgent need is established a kind of same
When detect the high-flux detection methods of more serotypes.Recent U.S. CDC uses liquid phase chip reagent box technical method, by right
The detection of O-antigen of salmonella, H antigen, AT antigen gene, the xMAP for developing 90% common Salmonella serogroup of covering are husky
Door Salmonella serotype fast typing kit, can obtain the serotype of salmonella in 4 hours, be the pre- of salmonella infection
Anti- and control provides strong tool.
Summary of the invention
For above-mentioned technical problem in the prior art, the present invention provides a kind of Streptococcus suis Serotypes liquid phase cores
Piece kit, this Streptococcus suis Serotypes liquid phase chip reagent box solve detection pig hammer in the prior art
It is the method heavy workload of bacterium Serotypes, at high cost, and the technical problem of result inaccuracy.
The invention reside in a kind of Streptococcus suis Serotypes liquid phase chip reagent box is provided, contain:
1) specific upstream primer probe, the sequence such as SEQ ID NO:41 of the upstream primer probe of the specificity
Shown in~60；
2) specific downstream primer probe, the sequence of the downstream primer probe of the specificity be SEQ ID NO:2,
SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14,
SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:
26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID
Shown in NO:38, SEQ ID NO:40, one section of biology added at 5 ' ends of any one of specific downstream primer probe
Element；
3) specific microballoon, the sequence of the specific microballoon is as shown in SEQ ID NO:61-80.
Further, design has one section of ATG label mutually complementary with microballoon in any one specific forward primer probe
And separately with 12 C.
Further, in the sequence of any one of specific microballoon and corresponding specific forward primer probe
TAG label is mutually complementary.
It further, also include hybrid night, Streptavidin (SAPE).
It further, also include multiplex PCR Mix 1, multiplex PCR Mix 2, multiplex PCR Mix 3, multiplex PCR Mix
4 and Streptococcus suis Serotypes liquid phase chip reagent box operation instructions.
Working principle of the present invention is: according to the difference of Streptococcus suis different serotypes capsule gene (Cps Gene), from
20 specific primers for Streptococcus suis specific serotype parting are designed on NCBI-Gene Bank；In specificity
Trip primer 5 ' end plus the preceding paragraph TAG sequence, centre are used 12 carbon bridges as termination signal, are added at the end of specificity downstream primer 5 '
Upper biotin (Biotin)；Select the microballoon mutually complementary with its TAG label (having Anti-TAG sequence)；Drawn with specific probe
Object goes amplification sample to be examined, and the PCR product expanded is then taken to hybridize with microballoon；Upper machine carries the specific probe of microballoon
By flow cytometer under the power of sheath fluid flowing, while machine issues twice laser, and red laser passes through to coding microball
Identification distinguish the number of microballoon, the biotin that green laser is used to identify that downstream primer 5 ' links is fixed as reporter gene
Amount finally obtains a MFI value (using 300 as threshold value).
Kit for Streptococcus suis Serotypes liquid-phase chip of the invention is a kind of detection technique of high throughput,
Collecting type cytometer technologies, fluorescence-encoded micro-beads, laser, Digital Signal Processing and traditional biochemical technology are a kind of in one
Multi-functional analysis platform.Liquid-phase chip technology accuracy and specificity are high, and the coincidence rate with serum agglutination and regular-PCR is
100%, while liquid-phase chip technology detection cycle is short, this kit can be completed in 3-4 hours to 20 kinds of serotype pig hammers
The screening of bacterium serotype；Compared with regular-PCR detection, not only detection time is short for this kit, and avoids harm of the EB to people,
High sensitivity, flux is big, and minimal detectable concentration can reach 1pg/ μ L, can be completed at the same time the detection of 20 kinds of serotype.This kit
It is combined as probe with specific microballoon using multiple PCR products, improves the specificity of detection, by flow cytometer and sharp
Optical scanning technology finally provides a MFI value, qualitative can not only can also carry out relative quantification to sample.Using the reagent
Box carries out Serotypes to clinical Streptococcus suis, has for the detection, epidemiological survey and risk assessment of Streptococcus suis
Important application value.
The present invention is compared with prior art, and technological progress is significant.Pig hammer is carried out using kit of the invention
When the liquid-phase chip of bacterium Serotypes, has many advantages, such as that quick, high-throughput, susceptibility is high, high specificity, greatly shorten
The application prospect of the kit is opened up experimental period.
Detailed description of the invention
Fig. 1 is the schematic diagram that liquid-phase chip instrument reads data.
Fig. 2 is the result expanded with the septuple PCR of this experimental design to Streptococcus suis serotype 1,2,3,7,9,11,16.
Fig. 3 is the result that the five weight PCR of this experimental design expand Streptococcus suis serotype 5,8,12,19,25.
Fig. 4 be this experimental design two Quadruple- PCRs respectively to Streptococcus suis serum 21,27,28,30 and serotype 13,
15, the result of 23,31 amplifications.
Fig. 5 is to verify liquid-phase chip to the minimal detectable concentration of Streptococcus suis, when detectable concentration is diluted to 105fg/ μ L,
The MFI that liquid-phase chip is shown is 53.
Fig. 6 is the amplification using traditional PCR method to 96 dilution gradients of type DNA of serotype, shows lowest detection
Concentration is 10.5pg/ μ L.
Fig. 7 is the schematic diagram and principle of specific probe and microballoon coupling
Fig. 8 is taken by two beam laser to microsphere surface when passing through flow cytometer under promotion of the microballoon in flowing sheath fluid
The information of band is scanned and reads.
Fig. 9 is specific PCR amplified production under suitable hybridization temperature, is coupled with specific microballoon.
Figure 10 is staphylococcus, the Escherichia coli, haemophilus parasuis, pleura lung for going verifying to be clinically separated with this kit
MFI value is below 300 as the result is shown for scorching Actinobacillus, Pasteurella, Streptococcus suis (respectively corresponding sample 1-6).
Figure 11 is to be clinically separated the result that pig streptococcus bacterial strain carries out serotype screening to 20 plants with this kit.
Specific embodiment
The following example is further intended to description invention, rather than limitation invention in any way, without departing substantially from the present invention
Spirit and principle under, ability made for the present invention or those of ordinary skill's any change easy to accomplish fall within this hair
Within bright pending interest field.
1 reagent box design principle of embodiment
The present invention is first according to the difference of Streptococcus suis different serotypes capsule gene (Cps Gene), from NCBI-Gene
20 NO:1~40 specific primer SEQ ID for Streptococcus suis specific serotype parting are designed on Bank；It is protecting
Card primer specificity and without dimer influence premise by 0 serotype of streptococcus suis 2 be divided into four groups of multiplex PCR systems (by
The reference culture that OIE streptococcus reference laboratory give, see Table 1 for details), first group is serotype 1,2,3,7,9,11,16, sees figure
2；Second group is serotype 5,8,12,19,25, sees Fig. 3；Third group is serotype 21,27,28,30, sees Fig. 4；4th group is blood
Clear type 13,15,23,31, is shown in Fig. 4；Specific forward primer 5 ' end plus the preceding paragraph TAG sequence, centre use 12 carbon bridges as
Termination signal is shown in SEQ ID 41~60 at the end of specificity downstream primer 5 ' plus biological (Biotin)；Selection is marked with its TAG
The complementary microballoon of label phase (having Anti-TAG sequence, SEQ ID 61~80) goes amplification sample to be examined with specific probe primer,
Then the PCR product after taking amplification hybridizes with microballoon (see Fig. 7)；Mixed liquor after hybridization is put into liquid-phase chip instrument and is read
Number, the specific probe for carrying microballoon passes through flow cytometer under the power that sheath fluid flows, while machine issues twice laser,
Red laser is used to identify what downstream primer 5 ' linked by distinguishing the number of microballoon, green laser to the identification of coding microball
Biotin is quantitative (see Fig. 1) as reporter gene, finally obtains a MFI value (using 300 as threshold value).
1 kit of table establishes the reference culture information that multiple PCR method is used
2 kit forms of embodiment
Multiplex PCR reagent: 24.5 μ L of PCR system
Combination 1 (includes 12.5 μ L of multiplex PCR Mix, the primer of serotype 1,2,3,7,9,11,16 each 0.8 μ L, ddH2O
Mend to 24.5 μ L), combination 2 (include 12.5 μ L of multiplex PCR Mix, each 0.8 μ L of the primer of serotype 5,8,12,19,25,
ddH2O mend to 24.5 μ L), combination 3 (comprising 12.5 μ L of multiplex PCR Mix, each 0.8 μ L of the primer of serotype 21,27,28,30,
ddH2O mend to 24.5 μ L), combination 4 (comprising 12.5 μ L of multiplex PCR Mix, each 0.8 μ L of the primer of serotype 13,15,23,31,
ddH2O is mended to 24.5 μ L).
Liquid-phase chip reagent: 90 μ L of reaction system, wherein the reaction microballoon 1 μ L comprising each coding, 1 × Tm are mended to 20 μ L,
70 μ LSAPE dilutions (69.3 1 × Tm of μ L and 0.7 μ LSAPE).
3 kit application method of embodiment
One, multiplex PCR
Reaction system is 25 μ L, 24.5 μ L multiplex PCR reagents (combination 1,2,3,4) and 0.5 μ L nucleic acid to be checked (10ng/ μ
L)。
Operating procedure: after sample nucleic acid to be checked is diluted according to above-mentioned requirements, it is more that 4 combinations in kit are separately added into
In weight PCR.
Reaction condition: 95 DEG C of 3min；95℃20s；60℃1min；72℃90s；35 circulations, 72 DEG C of extension 10min.
Two, hybridize
1. each coding microball participates in reaction (i.e. 1 μ L) with 2500 microballoons, 20 μ L are complemented to 1 × Tm；
2. the product after the 2 above-mentioned PCR amplifications of μ L is added, ddH is used2O is adjusted to 25 μ L of final volume；
3. SAPE is diluted to 8 μ g/ml with 1 × Tm hybridization buffer, and 70 μ L are added to each reacting hole, softly mix
It is even；
4. being put into PCR instrument, use 40 DEG C of hybridization 35min (see Fig. 9).
Three, liquid-phase chip instrument reads result
First machine should be cleaned before upper machine, and laser is calibrated and verified；90 μ L of liquid-phase chip reagent is taken, is added
Enter hybridization solution n μ L (2≤n≤5) to be checked, uses ddH2O is mended to 95 μ L of final volume；Liquid-phase chip instrument is every to 100 by 2 beam laser
Kind coding microball carries out laser scanning surface (see Fig. 8), reads 100 microsphere surface fluorescence values and then takes median, finally obtains MFI
Value (being threshold value with 300, be greater than 300 for the positive, be judged to feminine gender lower than 300).
4 kit specific test of embodiment
Take staphylococcus, Escherichia coli, haemophilus parasuis, Actinobacillus pleuropneumoniae, Pasteurella, Streptococcus suis
Each 0.5 μ L of nucleic acid of 6 control samples such as (corresponding sample 1-6) is that template carries out 4 groups of multiplexed PCR amplifications, hybridization, liquid-phase chips
Instrument reads the detection of MFI value, while setting negative control group.
Multiplexed PCR amplification condition: 95 DEG C, 3min；95 DEG C, 20s；60℃1min；72℃90s；35 circulations；72℃
10min；
Hybridization reaction condition: 40 DEG C, 35min；
Liquid-phase chip reaction condition: the copper sheet in liquid-phase chip instrument is heated to 40 DEG C in advance, the premixed liquid hybridized is answered
It puts into liquid-phase chip instrument rapidly, clicks " Run " operation.
Liquid-phase chip instrument is positive findings the result shows that only Streptococcus suis has MFI value (1565), and control group grape ball
Bacterium, Escherichia coli, haemophilus parasuis, Actinobacillus pleuropneumoniae, Pasteurella MFI value be below 300, be negative findings
(see Figure 10).
As a result: high specificity
5 kit sensitivity tests of embodiment
The DNA of 9 type of Streptococcus suis serotype is extracted, and carries out the measurement of DNA concentration, DNA concentration is 10.5ng/ μ L.
The DNA of extraction is subjected to 10 times of dilutions, dilutes 6 titres altogether, concentration is respectively 1.05ng/ μ L, 105pg/ μ L, 10.5pg/ μ
L, 1.05pg/ μ L, 105fg/ μ L, 10.5fg/ μ L, multiplexed PCR amplification hybridize with corresponding microballoon after the completion, then pass through liquid phase
Chip instrument reads data, and as DNA concentration 105fg/ μ L, MFI value is lower than threshold value 300 as the result is shown, so showing kit to pig
Streptococcus nucleic acid minimal detectable concentration is 1.05pg/ μ L (result is shown in Fig. 5).Race PCR is gone to carry out with above-mentioned 6 dilution gradient DNA
Susceptibility comparison, does not have band when DNA is (the 4th hole) 1.05pg/ μ L as the result is shown, is detailed in Fig. 6.
As a result: sensitivity is up to 1.05pg/ μ L
The detection of 6 serum agglutination of embodiment
Totally 20 plants of Streptococcus suis obtained from District of Shanghai separation are selected, number 1-20#；A small amount of point is picked with oese
From bacterial strain on sheep blood plate, 37 DEG C of culture 18-24h；Oese picks the sun of a small amount of bacterium colony Yu each serotype Streptococcus suis
Property serum 15 μ L (standard serum of Denmark's purchase) on clean slide, mixing；And positive and negative control is set；The blood in 4min
It clears out existing agglutination and is judged to the positive, no agglutination phenomenon is judged to feminine gender.
As a result: in 20 plants of isolated strains, thering are 13 plants to be aggregated, see Table 2 for details.
Table 2 is clinically separated the result that pig streptococcus bacterial strain is screened to 20 plants by serum agglutination method
The detection of 7 liquid-phase chip of embodiment
1. multiplex PCR selects totally 20 plants of Streptococcus suis obtained from District of Shanghai separation, number is 1-20# (solidifying with serum
Collection detection test sample is consistent), nucleic acid DNA is extracted, 4 groups of multi-PRC reactions: 95 DEG C of 3min are carried out；95℃20s；60℃90s；
72℃90s；35 circulations, 72 DEG C of extension 10min.
2. hybridization, 1 μ L of microballoon complement to 20 μ L with 1 × Tm；2 μ L of PCR product is added, uses ddH2O is adjusted to 25 μ of final volume
L；SAPE is diluted to 8 μ g/ml with 1 × Tm hybridization buffer, and 70 μ L are added to each reacting hole, soft mixing；It is put into PCR
Instrument, 40 DEG C of hybridization 35min.
3. liquid-phase chip instrument reads data.
The results show that there is 13 plants of separation bacterium MFI to be greater than 300, the serotype detected is 2 (sample of serotype respectively
1/6/10), serotype 3 (sample 3), serotype 5 (sample 11), serotype 7 (sample 4/7/9), serotype 9 (sample 2/8), blood
Clear type 19 (sample 5) serotype 21 (sample 12), serotype 27 (sample 13), are detailed in Figure 11.
By embodiment 6 and embodiment 7 it may be concluded that the knot that the test result and liquid-phase chip of serum agglutination detect
Fruit is consistent.
All references mentioned in the present invention is incorporated herein by reference, independent just as each document
It is incorporated as with reference to such.In addition, it should also be understood that, after reading the above teachings of the present invention, those skilled in the art can
To make various changes or modifications to the present invention, such equivalent forms equally fall within model defined by the application the appended claims
It encloses.
Claims (2)
1. a kind of Streptococcus suis Serotypes liquid phase chip reagent box, it is characterised in that it contains:
1) upstream primer of specific upstream primer, the specificity includes sequence as shown in NO:41 ~ 60 SEQ ID
Primer；
2) downstream primer of specific downstream primer, the specificity includes sequence such as SEQ ID NO:2, SEQ ID NO:
4, SEQ ID NO:6, SEQ ID NO:8, SEQIDNO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16,
SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:
28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID
Primer shown in NO:40 adds biotin at 5 ' ends of any one of specific downstream primer；
3) specific microballoon, the specific microballoon includes sequence probe as shown in NO:61 ~ 80 SEQ ID.
2. a kind of Streptococcus suis Serotypes liquid phase chip reagent box according to claim 1, it is characterised in that: include
1 × Tm hybridization solution, Streptavidin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510954076.2A CN105385766B (en) | 2015-12-17 | 2015-12-17 | A kind of Streptococcus suis Serotypes liquid phase chip reagent box |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510954076.2A CN105385766B (en) | 2015-12-17 | 2015-12-17 | A kind of Streptococcus suis Serotypes liquid phase chip reagent box |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105385766A CN105385766A (en) | 2016-03-09 |
| CN105385766B true CN105385766B (en) | 2019-01-29 |
Family
ID=55418541
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510954076.2A Active CN105385766B (en) | 2015-12-17 | 2015-12-17 | A kind of Streptococcus suis Serotypes liquid phase chip reagent box |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105385766B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105648101A (en) * | 2016-03-28 | 2016-06-08 | 上海市动物疫病预防控制中心 | Liquid-phase chip kit for screening seven virulence genes of streptococcus suis |
| CN106191313A (en) * | 2016-07-21 | 2016-12-07 | 上海市动物疫病预防控制中心 | A kind of six kinds of pig breeding dysfunction virus liquid phase chip agent boxes |
| CN107312873B (en) * | 2017-06-28 | 2018-07-24 | 广东省实验动物监测所 | A kind of multiple liquid phase genetic chip detection primer, kit and method of 5 kinds of Respiratory Tract of Mice cause of diseases of quick differentiation |
| CN112280880A (en) * | 2020-11-20 | 2021-01-29 | 中国疾病预防控制中心传染病预防控制所 | Primer combination for identifying 17 new cps of streptococcus suis by liquid chip multiplex PCR and detection kit thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101445831A (en) * | 2008-12-23 | 2009-06-03 | 广州益善生物技术有限公司 | FSHR gene mutation detection liquid phase chip and detection method thereof |
| CN103205509A (en) * | 2013-04-23 | 2013-07-17 | 中国检验检疫科学研究院 | High-flux non-diagnostic detection method for 13 respiratory viruses based on novel suspension chip technology |
-
2015
- 2015-12-17 CN CN201510954076.2A patent/CN105385766B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101445831A (en) * | 2008-12-23 | 2009-06-03 | 广州益善生物技术有限公司 | FSHR gene mutation detection liquid phase chip and detection method thereof |
| CN103205509A (en) * | 2013-04-23 | 2013-07-17 | 中国检验检疫科学研究院 | High-flux non-diagnostic detection method for 13 respiratory viruses based on novel suspension chip technology |
Non-Patent Citations (1)
| Title |
|---|
| Fifteen Streptococcus suis serotypes identified by multiplex PCR;Anusak Kerdsin;《Journal of Medical Microbiology》;20120823;第61卷;1669-1672 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105385766A (en) | 2016-03-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| O'Sullivan | Methods for analysis of the intestinal microflora | |
| CN105385766B (en) | A kind of Streptococcus suis Serotypes liquid phase chip reagent box | |
| CN103074451B (en) | Kit for synchronously detecting twenty-two respiratory tract pathogens and detection method of kit | |
| CN103911447B (en) | Detect plasmodial primer, probe and method | |
| CN106868167A (en) | Primer, probe and kit for field quick detection Mycoplasma bovis | |
| CN107868834A (en) | The real-time quantitative PCR detection primer group and method of a kind of acipenser dabryanus sexual gland difference expression gene | |
| CN109913565A (en) | A kind of kit for detecting vibrio parahaemolytious, primer pair, probe and method | |
| Namsolleck et al. | Molecular methods for the analysis of gut microbiota | |
| CN101613751A (en) | The PCR kit for fluorescence quantitative of rapid detection toxoplasma gondii | |
| CN103898240B (en) | Detect the primer of dengue virus and somatotype thereof, probe and method | |
| CN111088380A (en) | Brucella LF-RPA detection primer, probe and detection kit | |
| CN106222256A (en) | A kind of Multiple immunizations fluorescence analysis method detecting chicken virus mycoplasma and chicken Mycoplasma synoviae | |
| CN109402274A (en) | A kind of fluorescent quantitative RT-PCR method identifying A type and Type B ox source pasteurella multocida | |
| CN106011313B (en) | A kind of the multi-fluorescence immunoassay method and reagent of quick differentiation ILTV, IBV, MG and MS | |
| CN107190103A (en) | Multiple PCR primer group, kit and the method for three kinds of fishes virus are detected simultaneously | |
| CN104975077B (en) | Pig source eperythrozoon fluorescent quantificationally PCR detecting kit and its application | |
| WO2015020671A1 (en) | Multiplex amplification reaction method for determination of campylobacter jejuni penner/capsule type | |
| CN109097488A (en) | For synchronizing nucleic acid, method and the kit of five kinds of dog diarrhea virus of detection and identification | |
| Koekemoer | Serotype-specific detection of African horsesickness virus by real-time PCR and the influence of genetic variations | |
| CN105154559A (en) | Specific nucleotide for vibrio parahaemolyticus K36, K37 and K68 and application thereof | |
| CN107937595A (en) | A kind of method, primer and probe and application that feed transgenic corn IE034 is quantitatively detected based on ddPCR | |
| CN102851395B (en) | Liquid-phase chip method used for detecting infectious laryngotracheitis virus | |
| CN105648101A (en) | Liquid-phase chip kit for screening seven virulence genes of streptococcus suis | |
| EP2539458B1 (en) | Multiplex amplification reaction method for determination of campylobacter jejuni penner/capsule type | |
| CN108642190A (en) | Forensic medicine composite detection kit based on 14 autosome SNP genetic markers |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |