CN107287320A - The LAMP detections of GAPDH genes are combined and kit with primer - Google Patents
The LAMP detections of GAPDH genes are combined and kit with primer Download PDFInfo
- Publication number
- CN107287320A CN107287320A CN201710567594.8A CN201710567594A CN107287320A CN 107287320 A CN107287320 A CN 107287320A CN 201710567594 A CN201710567594 A CN 201710567594A CN 107287320 A CN107287320 A CN 107287320A
- Authority
- CN
- China
- Prior art keywords
- primer
- sequence
- seq
- lamp
- gapdh
- 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
- C12Q1/6844—Nucleic acid amplification reactions
-
- 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
-
- 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
- C12Q2600/166—Oligonucleotides used as internal standards, controls or normalisation probes
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Combined the invention provides a kind of GAPDH genetic tests with primer and kit, using sequence shown in SEQ ID No.1 as target sequence, design primer combination, the primer combination includes F3, B3, FIP, BIP, LF, LB.The primer sets are shared in LAMP detection kit, as a result show that primer combination does not occur non-specific amplification and primer inside primer and the non-specific binding amplification of template, the internal reference system of quality control of humanization sample LAMP methods amplification has been successfully established, preliminary basis is established in clinical extensive use for LAMP methods in future.
Description
Technical field
Combined the present invention relates to a kind of GAPDH genetic tests with primer and kit, belong to biological technical field.
Background technology
GAPDH or G3PDH are glyceraldehyde-3-phosphate dehydrogenase (glyceraldehyde-3-phosphate
Dehydrogenase english abbreviation).The enzyme is an enzyme in glycolysis reaction, is made up of 4 30-40kDa subunit,
Molecular weight 146kDa.The enzyme gene is house keeper (house keeping) gene, almost in a organized way in all high level expressions,
Protein expression amount in allogenic cell or tissue is usually constant, and not by containing part recognition site, Buddhist ripple
The influence of the inducing substance of fat etc. and keep constant, therefore be widely used as PCR, extracting total RNA, poly (A)+RNA,
The internal reference of the standardization of the experimental implementations such as Western blot.
With the development of laboratory medicine, some Protocols in Molecular Biologies, such as PCR, strand displacement amplification, medical test with
It played an important role in terms of diagnosis.However, due to Protocols in Molecular Biology detect nucleic acid to instrument, equipment requirement compared with
Height, operation sequence is complicated, limits it as the development and popularization of fast diagnosis method, loop-mediated isothermal amplification technique (loop
Mediated isothermal amplification, LAMP) solve the difficulty such as equipment costliness, program complexity in diagnostic nucleic acid
Topic.
LAMP reactions only need a kind of archaeal dna polymerase (such as Bst, Gsp) for having strand-displacement activity and individual for target gene 6-8
The 4-6 bars primer (F3/B3 and FIP/BIP are required, and LF/LB is optional) of region design just can be under 60 DEG C~66 DEG C of steady temperature
Realize that the target gene for being more than 109 times in dozens of minutes is expanded, its lower bound detected up to 10 copies/microlitre, by high expansion
Increasing Efficiency and relatively low instrument requirements, the technology are gradually applied to pathogen identification, biological marker analyte detection, sex identification
Deng, it should be noted that：The laboratories detection field of LAMP technology quick detection, field detection and resource-constrained by bed
There is spacious wealthy prospect.
Limit to however, the in vitro practical application of LAMP technology is relative in recent years, be in progress relatively slow, this seminar
Find that limiting its wide variety of major reason there are two according to the research of early stage, be specially：(1) current lamp systems are difficult
To realize high-throughout detection truly, await further perfect；(2) although nothing in LAMP systems, detection process
Heating and cooling are needed, reaction speed can be greatly speeded up, efficient, quick detection is realized, but compared with traditional real-time PCR methodology,
The less stable of LAMP systems, often occurs and contains excessive PCR inhibitor in failure or sample because sample nucleic acid is extracted
Caused amplification failure, so as to cause the appearance of false negative result, therefore, in order to avoid false negative, improves testing result credible
Degree is, it is necessary to introduce internal reference amplification system, to sample process and the progress effective monitoring of the process of detection.
The content of the invention
The technical problem to be solved in the present invention is, in view of the shortcomings of the prior art, proposing a kind of GAPDH genetic tests primer
Combination and kit.For people source biological specimen, with one section of highly conserved target sequence on people's GAPDH genes, primer is designed,
Establish GAPDH-LAMP detection architectures and optimize, be successfully established the internal reference Quality Control body of humanization sample LAMP methods amplification
System, preliminary basis is established for LAMP methods in future in clinical extensive use.
The purpose of the present invention is achieved through the following technical solutions：
In a first aspect, the invention provides a kind of target sequence for people's GAPDH genetic tests, the sequence such as SEQ ID
Shown in No.1.
Second aspect, the invention provides a kind of primer combination designed according to foregoing target sequence, the primer combination bag
Include F3, B3, FIP, BIP, LF, LB；The sequence of the primers F 3 is as shown in SEQ ID No.2, primer B3 sequence such as SEQ ID
Shown in No.3, primers F IP sequence is as shown in SEQ ID No.4, and primer BIP sequence is as shown in SEQ ID No.5, primer LF
Sequence as shown in SEQ ID No.6, primer LB sequence is as shown in SEQ ID No.7.
The third aspect, the invention provides a kind of application of primer combination in detection GAPDH genes.
Fourth aspect, the invention provides a kind of LAMP kit for people's GAPDH genetic tests, including amplified reaction
System, the cumulative volume of the amplification reaction system is 25 μ L, including：The μ L of 2 × reaction buffer 12.5, primers F 3：1 μ L, primer
B3：1 μ L, primers F IP：0.5 μ L, primer BIP：1 μ L, primer LF：1 μ L, primer LB：The μ L of 1 μ L, Bst archaeal dna polymerase 1, calcium is yellow
Green plain 1 μ L, the μ L of deionized water 3.0, the μ L of human gene group DNA's template 2.
Preferably, the primers F 3 and primer B3 final concentration of 0.2 μm of ol/L, primers F IP and primer BIP final concentration
For 1.6 μm of ol/L, primer LF and primer LB final concentration of 0.8 μm of ol/L.
Preferably, the concentration of the Bst archaeal dna polymerases is 8U.
5th aspect, the invention provides a kind of method for detecting people's GAPDH genes, comprises the following steps：Using foregoing
LAMP kit carry out LAMP reactions, reaction terminate after, judged by the color change of reaction solution, the color of reaction solution
From it is original it is faint yellow be changed into green, that is, be determined as reacting positive.
Preferably, the condition of the LAMP reactions is：65 DEG C of reaction temperature, reaction time 60min.
Compared with prior art, the present invention has following beneficial effect：
With one section of highly conserved target sequence on people's GAPDH genes, primer is designed, the primer sets are shared in LAMP detections
In kit, as a result show that primer combination does not occur non-specific amplification and primer inside primer and the non-specific knot of template
Amplification is closed, GAPDH-LAMP detection architectures is thereby establish and optimizes, the amplification of humanization sample LAMP methods has been successfully established
Internal reference system of quality control, establish preliminary basis in clinical extensive use for LAMP methods in future.
Brief description of the drawings
By reading the detailed description made with reference to the following drawings to non-limiting example, further feature of the invention,
Objects and advantages will become more apparent upon：
Fig. 1, the testing result schematic diagram for the embodiment of the present invention；Wherein, A1:Clinical wildtype genomic DNA is template,
A2:108Copies/ml plasmid sample, A3:107Copies/ml plasmid sample, A4:106Copies/ml plasmid sample,
A5:105Copies/ml plasmid sample, A6:104Copies/ml plasmid sample, A7：103Copies/ml plasmid sample,
A8：Negative control sample.
Embodiment
With reference to specific embodiment, the present invention is described in detail.Following examples will be helpful to the technology of this area
Personnel further understand the present invention, but the invention is not limited in any way.It should be pointed out that to the ordinary skill of this area
For personnel, without departing from the inventive concept of the premise, some changes and improvements can also be made.These belong to the present invention
Protection domain.
The GAPDH-LAMP detection architectures of embodiment 1 are set up
(1) with one section of highly conserved target sequence on people's GAPDH genes, design primer, the long 185bp of target sequence,
Specially:
GAACCAGCACCGATCACCTCCCATCGGGCCAATCTCAGTCCCTTCCCCCCTACGTCGGGGCCCACACGCTCGGTGCG
TGCCCAGTTGAACCAGGCGGCTGCGGAAAAAAAAAAGCGGGGAGAAAGTAGGGCCCGGCTACTAGCGGTTTTACGGG
CGCACGTAGCTCAGGCCTCAAGACCTTGGGC(SEQ ID No.1)
The primer sequence is as shown in table 1.
Table 1
(2) kit used includes amplification reaction system, and the cumulative volume of the amplification reaction system is 25 μ L, and it includes
The μ L of 2 × reaction buffer (RM) 12.5, primers F 3：1 μ L (0.2 μm of ol/L of final concentration), primer B3：1 μ L (0.2 μm of ol/ of final concentration
L), primers F IP：0.5 μ L (1.6 μm of ol/L of final concentration), primer BIP：1 μ L (1.6 μm of ol/L of final concentration), primer LF：1 μ L are (eventually
0.8 μm of ol/L of concentration), primer LB：1 μ L (0.8 μm of ol/L of final concentration), the μ L (8U) of Bst archaeal dna polymerases 1, calcein (FD) 1
μ L, the μ L of deionized water 3.0, the μ L of DNA profiling 2.
LAMP reactions are carried out using mentioned reagent box, LAMP reaction conditions are：65 DEG C, 60min；Equipment used is common
PCR instrument or constant-temperature metal bath etc. can stablize the equipment for providing 65 DEG C of constant temperature；Then carry out visualization interpretation is to identify sample
No is the positive, and as a result judgement is specially：After reaction terminates, the color of reaction solution from it is original it is faint yellow be changed into green, that is, judge
For reacting positive.
The LAMP detections of the GAPDH genes of embodiment 2
The TA cloned plasmids containing target sequence (SEQ ID No.1) are built, the double sample of gradient concentration, specific concentration is prepared
For 103copies/ml、104copies/ml、105copies/ml、106copies/ml、107copies/ml、108copies/
Ml, the GAPDH-LAMP reaction systems set up with embodiment 1 are detected, can be detected, as a result as shown in Figure 1.Wherein,
A1：Human peripheral genomic DNA is template, A2：108Copies/ml plasmid sample, A3：107Copies/ml plasmid sample
This, A4：106Copies/ml plasmid sample, A5：105Copies/ml plasmid sample, A6：104Copies/ml plasmid sample
This, A7：103Copies/ml plasmid sample, A8：Negative control sample.After the system of A1-A7 samples is expanded through 60min,
Greening becomes cloudy, and positive findings is presented；A8 is negative control, after being expanded through 60min, is not any change, still faint yellow clarification,
Negative findings is presented.
In summary, the present invention has following beneficial effect：
With one section of highly conserved target sequence on people's GAPDH genes, primer is designed, the primer sets are shared in LAMP detections
In kit, as a result show that primer combination does not occur non-specific amplification and primer inside primer and the non-specific knot of template
Amplification is closed, GAPDH-LAMP detection architectures is thereby establish and optimizes, the amplification of humanization sample LAMP methods has been successfully established
Internal reference system of quality control, establish preliminary basis in clinical extensive use for LAMP methods in future.
The specific embodiment of the present invention is described above.It is to be appreciated that the invention is not limited in above-mentioned
Particular implementation, those skilled in the art can make a variety of changes or change within the scope of the claims, this not shadow
Ring the substantive content of the present invention.In the case where not conflicting, feature in embodiments herein and embodiment can any phase
Mutually combination.
SEQUENCE LISTING
<110>Cao monarch
<120>The LAMP detections of GAPDH genes are combined and kit with primer
<130> DAG29909
<160> 7
<170> PatentIn version 3.3
<210> 1
<211> 185
<212> DNA
<213> artificial sequence
<220>
<223> artificial sequence
<400> 1
gaaccagcac cgatcacctc ccatcgggcc aatctcagtc ccttcccccc tacgtcgggg 60
cccacacgct cggtgcgtgc ccagttgaac caggcggctg cggaaaaaaa aaagcgggga 120
gaaagtaggg cccggctact agcggtttta cgggcgcacg tagctcaggc ctcaagacct 180
tgggc 185
<210> 2
<211> 18
<212> DNA
<213> artificial sequence
<220>
<223> artificial sequence
<400> 2
gaaccagcac cgatcacc 18
<210> 3
<211> 17
<212> DNA
<213> artificial sequence
<220>
<223> artificial sequence
<400> 3
gcccaaggtc ttgaggc 17
<210> 4
<211> 38
<212> DNA
<213> artificial sequence
<220>
<223> artificial sequence
<400> 4
aactgggcac gcaccgagct cccatcgggc caatctca 38
<210> 5
<211> 35
<212> DNA
<213> artificial sequence
<220>
<223> artificial sequence
<400> 5
accaggcggc tgcggaaagc tacgtgcgcc cgtaa 35
<210> 6
<211> 17
<212> DNA
<213> artificial sequence
<220>
<223> artificial sequence
<400> 6
acgtaggggg gaaggga 17
<210> 7
<211> 23
<212> DNA
<213> artificial sequence
<220>
<223> artificial sequence
<400> 7
aaaaaaagcg gggagaaagt agg 23
Claims (8)
1. a kind of target sequence for people's GAPDH genetic tests, it is characterised in that the sequence is as shown in SEQ ID No.1.
2. a kind of primer combination of the design of the target sequence according to claim 1, it is characterised in that the primer combination includes
F3、B3、FIP、BIP、LF、LB；The sequence of the primers F 3 is as shown in SEQ ID No.2, primer B3 sequence such as SEQ ID
Shown in No.3, primers F IP sequence is as shown in SEQ ID No.4, and primer BIP sequence is as shown in SEQ ID No.5, primer LF
Sequence as shown in SEQ ID No.6, primer LB sequence is as shown in SEQ ID No.7.
3. a kind of application of primer combination according to claim 2 in detection GAPDH genes.
4. a kind of LAMP kit for people's GAPDH genetic tests, it is characterised in that including amplification reaction system, the expansion
The cumulative volume for increasing reaction system is 25 μ L, including：The μ L of 2 × reaction buffer 12.5, primers F 3：1 μ L, primer B3：1 μ L, primer
FIP：0.5 μ L, primer BIP：1 μ L, primer LF：1 μ L, primer LB：1 μ L, BstDNA polymerase 1 μ L, the μ L of calcein 1, go from
The sub- μ L of water 3.0, the μ L of human gene group DNA's template 2.
5. the LAMP kit according to claim 4 for people's GAPDH genetic tests, it is characterised in that the primer
F3 and primer B3 final concentration of 0.2 μm of ol/L, primers F IP and primer BIP final concentration of 1.6 μm of ol/L, primer LF and draw
Thing LB final concentration of 0.8 μm of ol/L.
6. the LAMP kit according to claim 4 for people's GAPDH genetic tests, it is characterised in that the Bst
The concentration of archaeal dna polymerase is 8U.
7. a kind of method for detecting people's GAPDH genes, it is characterised in that comprise the following steps：Using described in claim 4
LAMP kit carries out LAMP reactions, after reaction terminates, is judged by the color change of reaction solution, the color of reaction solution by
Originally faint yellow is changed into green, that is, is determined as reacting positive.
8. the method for detection people's GAPDH genes according to claim 7, it is characterised in that the condition of the LAMP reactions
For：65 DEG C of reaction temperature, reaction time 60min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710567594.8A CN107287320A (en) | 2017-07-12 | 2017-07-12 | The LAMP detections of GAPDH genes are combined and kit with primer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710567594.8A CN107287320A (en) | 2017-07-12 | 2017-07-12 | The LAMP detections of GAPDH genes are combined and kit with primer |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107287320A true CN107287320A (en) | 2017-10-24 |
Family
ID=60100745
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710567594.8A Pending CN107287320A (en) | 2017-07-12 | 2017-07-12 | The LAMP detections of GAPDH genes are combined and kit with primer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107287320A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107988358A (en) * | 2017-12-28 | 2018-05-04 | 湖北工业大学 | A kind of the internal reference amplimer composition and its amplification system of detection NPPA gene mutations c.T413C |
| CN110863051A (en) * | 2019-12-13 | 2020-03-06 | 广州迈景基因医学科技有限公司 | Primer, system and kit for MET gene amplification detection |
| CN112266986A (en) * | 2020-12-22 | 2021-01-26 | 博奥生物集团有限公司 | Virus nucleic acid extraction or preservation reagent, primer probe combination, virus amplification reagent, kit and application thereof |
| US11788129B2 (en) | 2021-02-23 | 2023-10-17 | Industrial Technology Research Institute | Nucleic acid detection kit and nucleic acid detection method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1473202A (en) * | 2001-03-09 | 2004-02-04 | Ŧب������˾ | Methods and compositions for amplification of RNA sequences |
| CN1633500A (en) * | 2002-02-20 | 2005-06-29 | 希森美康株式会社 | Primers for nucleic acid amplification in detecting housekeeping gene mRNA and test method using these primers |
| WO2014060604A2 (en) * | 2012-10-20 | 2014-04-24 | Selfdiagnostics OÜ | Method and its compositions for detection of nucleic acid target from biological samples and body fluids |
| EP3098326A1 (en) * | 2015-05-27 | 2016-11-30 | Institut Pasteur | Isothermal amplification assay for rapid and accurate detection of hemorrhagic fever viruses in clinical samples |
-
2017
- 2017-07-12 CN CN201710567594.8A patent/CN107287320A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1473202A (en) * | 2001-03-09 | 2004-02-04 | Ŧب������˾ | Methods and compositions for amplification of RNA sequences |
| CN1633500A (en) * | 2002-02-20 | 2005-06-29 | 希森美康株式会社 | Primers for nucleic acid amplification in detecting housekeeping gene mRNA and test method using these primers |
| WO2014060604A2 (en) * | 2012-10-20 | 2014-04-24 | Selfdiagnostics OÜ | Method and its compositions for detection of nucleic acid target from biological samples and body fluids |
| EP3098326A1 (en) * | 2015-05-27 | 2016-11-30 | Institut Pasteur | Isothermal amplification assay for rapid and accurate detection of hemorrhagic fever viruses in clinical samples |
Non-Patent Citations (3)
| Title |
|---|
| RIEDER,M.J.等: "GenBank: AY340484.1，Homo sapiens glyceraldehyde-3-phosphate dehydrogenase (GAPD) gene, complete cds", 《GENBANK DATABASE》 * |
| 夏邦顺等主编: "《临床分子诊断学》", 31 July 2012, 中山大学出版社 * |
| 谭贵良等主编: "《现代分子生物学及组学技术在食品安全检测中的应用》", 30 June 2014, 中山大学出版社 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107988358A (en) * | 2017-12-28 | 2018-05-04 | 湖北工业大学 | A kind of the internal reference amplimer composition and its amplification system of detection NPPA gene mutations c.T413C |
| CN110863051A (en) * | 2019-12-13 | 2020-03-06 | 广州迈景基因医学科技有限公司 | Primer, system and kit for MET gene amplification detection |
| CN110863051B (en) * | 2019-12-13 | 2020-10-23 | 广州迈景基因医学科技有限公司 | Primer, system and kit for MET gene amplification detection |
| CN112266986A (en) * | 2020-12-22 | 2021-01-26 | 博奥生物集团有限公司 | Virus nucleic acid extraction or preservation reagent, primer probe combination, virus amplification reagent, kit and application thereof |
| US11788129B2 (en) | 2021-02-23 | 2023-10-17 | Industrial Technology Research Institute | Nucleic acid detection kit and nucleic acid detection method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110184329A (en) | A kind of one-step method nucleic acid detection method and kit based on CRISPR/Cas and constant-temperature amplification | |
| CN107287320A (en) | The LAMP detections of GAPDH genes are combined and kit with primer | |
| CN101712973B (en) | Reactive reagent of nucleic acid amplification by chain replacement at room temperature and nucleic acid amplification method at room temperature thereof | |
| CN109055498A (en) | MiRNA and/or biological micromolecule detection probe, detection method and kit based on the dendritic rolling ring responsive transcription of hypertree | |
| CN108531630A (en) | The streptococcic primer sets of detection B races include the primer liquid and kit of the primer sets and its application | |
| CN108913811A (en) | The rapid amplifying method of hepatitis B virus nucleic acid | |
| CN115029459A (en) | Kit for visually detecting Pasteurella multocida based on CRISPR-Cas12a and application | |
| CN104561360A (en) | TOP2A gene abnormality detection probe, kit and method | |
| CN106399536B (en) | Body fluid circulatory DNA quantitative detecting method and kit | |
| CN103276099B (en) | Primer and kit for fluorescent quatititive PCR (polymerase chain reaction) detection of helicobacter pylori | |
| CN105132571B (en) | Dissociated the method for mitochondrial DNA content using droplet type digitlization PCR detection peripheral bloods | |
| CN110093418B (en) | piRNA-54265 detection kit for early screening, diagnosis, curative effect and prognosis evaluation of colorectal cancer | |
| CN107338289A (en) | A kind of LAMP primer composition thing of lock nucleic acid modification and its application | |
| CN102181567A (en) | Real-time fluorescent quantitative PCR (polymerase chain reaction) kit for detecting candida glabrata | |
| CN103571939B (en) | Fluorescent quantitative polymerase chain reaction (PCR) method for detecting tubercle bacillus infection from clinical specimen | |
| CN114525349A (en) | Method for identifying semen by using digital PCR and special kit thereof | |
| CN116814789A (en) | System for cervical cancer high-grade lesion or cervical cancer diagnosis and related gene methylation qPCR detection kit | |
| CN104087665B (en) | A kind of universal primer and real-time fluorescence tHDA test kit detecting flavus and Aspergillus fumigatus | |
| CN105543346B (en) | A kind of Richettsia multiple fluorescence PCR detection method | |
| CN107326092A (en) | Applications and colorectal cancer detection kit of the related miRNA of colorectal cancer as biomarker | |
| CN104328210B (en) | The primer of the loop-mediated isothermal amplification detection method of AML1/ETO fusion gene and test kit | |
| CN111154836A (en) | Targeted nucleic acid capture and detection methods | |
| CN113337637A (en) | Primer group and kit for molecular detection of SARS-CoV-2 coronavirus | |
| CN109266786A (en) | African swine fever virus detection kit and detection method based on E184L gene | |
| CN111321239B (en) | LAMP primer group for detecting moniliforme and detection method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20171024 |