CN109337976A - For detect PIK3CD gene E1021K site mutation probe and primer combination and kit - Google Patents
For detect PIK3CD gene E1021K site mutation probe and primer combination and kit Download PDFInfo
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- CN109337976A CN109337976A CN201811584627.0A CN201811584627A CN109337976A CN 109337976 A CN109337976 A CN 109337976A CN 201811584627 A CN201811584627 A CN 201811584627A CN 109337976 A CN109337976 A CN 109337976A
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- probe
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- 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
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
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- 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
- C12Q2600/156—Polymorphic or mutational markers
Abstract
The present invention proposes a kind of saltant type for detecting mankind's PIK3CD gene C .3061G > A, P.E1021K site mutation and wild-type probe and primer combination.The nucleotide sequence of forward primer is as shown in SEQ ID NO:1, and the nucleotide sequence of reverse primer is as shown in SEQ ID NO:2；The probe includes saltant type probe, and the nucleotide sequence of the saltant type probe is as shown in SEQ ID NO:3.PIK3CD gene C .3061G > A that the present invention passes through the specificity that design is scientific and reasonable, P.E1021K detection primer and probe, preferably to realize the method detection based on quantitative fluorescent PCR, for method based on the quantitative fluorescent PCR realized on the basis of probe provided by the invention and primer on the basis of guaranteeing highly sensitive and high specific, experimental period is shorter, cost is lower.
Description
Technical field
The invention belongs to biomedicine technical field, more particularly to it is a kind of for detect mankind PIK3CD gene c.3061G >
The probe and primer of A, p.E1021K site mutation combine and kit.
Background technique
PI3K- δ overactivity syndrome is that a kind of autosomal dominant inheritance as caused by PIK3CD gene mutation is immunized
Defect disease.The disease is usually expressed as the defect of T cell and B cell differentiation and function.Main clinic symptoms have: childhood onset
Repeated respiratory infections, enlargement of lymph nodes, nodular lymphoid hyperplasia, splenomegaly, progressive lymphocyte is reduced, antibody response lacks
It falls into and CMV and/or EBV infects caused viremia virusemia.Although lymph node pathological change and nodular lymphoid hyperplasia are non-cancerous
(benign), but PI3K- δ overactivity syndrome also increases the risk to form cancer B cell lymphoma.The disease onset
Early, state of an illness weight, serious person can be fatal.And the disease can be by rapamycin treatment, effect is preferable.Therefore early diagnosis will be advantageous
In the treated as soon as possible of infant, patient vitals are saved.
It at present can be by PIK3CD full genome Sanger sequencing or high throughput for PI3K- δ overactivity syndrome
Sequencing technologies (Next generation Sequencing) carry out deep sequencing to PIK3CD gene and other genes, can obtain
Abrupt information in PIK3CD whole gene, but inevitably there are sequencing costs it is high, experimental period is long the disadvantages of.Early period
Research finds that c.3061G > A, p.E1021K are the mutantional hotspot of the disease, the Largest In China ancestor that Pediatrics magazine is reported
PI3K- δ overactivity syndrome patient is the point mutation.Therefore how to realize quickly detect this site can be rapid
Primary dcreening operation is carried out to the disease, shortens interval between diagnosis, reduce testing cost, it is difficult to become those skilled in the art's technology urgently to be resolved
Topic.
Summary of the invention
The present invention for the above technical issues, propose it is a kind of for detect mankind PIK3CD gene c.3061G > A,
The probe and primer of p.E1021K site mutation combine and kit so that PIK3CD gene c.3061G > A, the site p.E1021K
The detection sensitivity of mutation is higher, specific higher and favorable repeatability, while can reduce experimentation cost, realizes PIK3CD base
Because of c.3061G > A, the quick detection of p.E1021K site mutation, and it is prominent to distinguish wild type, heterozygous mutant and homozygosis
Modification.If there are somatic mutation or chimeras for the disease, its frequency of mutation also can detect.
One kind for detect mankind PIK3CD gene c.3061G > A, p.E1021K site mutation probe and primer combination,
The primer includes forward primer and reverse primer, and the nucleotide sequence of the forward primer is described as shown in SEQ ID NO:1
The nucleotide sequence of reverse primer is as shown in SEQ ID NO:2；The probe includes saltant type probe, the saltant type probe
Nucleotide sequence is as shown in SEQ ID NO:3.
Preferably, the molar ratio of the forward primer, reverse primer and the saltant type probe is 1:1:1.
Preferably, the probe further includes wild-type probe, the nucleotide sequence of the wild-type probe such as SEQ ID
Shown in NO:4.
Preferably, 5 ' ends of the saltant type probe and wild-type probe are marked with fluorophor, 3 ' ends, which are marked with, quenches
Go out group.
Preferably, the fluorophor is any one of FAM, CY5, HEX, VIC, ROX.
Wherein the fluorophor and quenching group can be screened based on actual using needs or cost consideration.Specifically
The saltant type probe fluorophor be FAM, quenching group BHQ1；The wild-type probe fluorophor is CY5, is quenched
The group that goes out is BHQ2.Or the fluorophor of the saltant type probe is CY5, quenching group BHQ2；The wild type
Fluorescence probe group is FAM, quenching group BHQ1.
Above-mentioned selected two groups of fluorophors and quenching group, realize PIK3CD gene c.3061G > A, p.E1021K
On the basis of site mutation quantitative detection, there is lower use cost.
Preferably, the molar ratio of the forward primer and reverse primer is 1:1, the wild-type probe and saltant type are visited
The molar ratio of needle is 1:1, and the molar ratio of the forward primer and the wild-type probe is (1-3): 1.
More preferably, the molar ratio of the forward primer, reverse primer, wild-type probe and saltant type probe is 1:1:
1:1。
One kind for detect mankind PIK3CD gene c.3061G > A, the kit of p.E1021K site mutation, including above-mentioned
For detect mankind PIK3CD gene c.3061G > A, p.E1021K site mutation probe and primer combination.
It is a kind of it is above-mentioned for detect mankind PIK3CD gene c.3061G > A, the probe and primer of p.E1021K site mutation
Combination preparation for detect mankind PIK3CD gene c.3061G > A, in the kit of p.E1021K site mutation and mutation rate
Application.
It is a kind of it is above-mentioned for detect mankind PIK3CD gene c.3061G > A, the probe and primer of p.E1021K site mutation
Combination preparation for screening PIK3CD gene c.3061G > A, p.E1021K site mutation correlation PI3K δ overactivity syndrome
Product in application.
Compared with prior art, the advantages and positive effects of the present invention are:
Provided by the present invention for detection mankind PIK3CD gene c.3061G > A, p.E1021K site mutation probe and draw
Object combination and kit, by the PIK3CD gene of design is scientific and reasonable specificity c.3061G > A, p.E1021K site mutation
Detection primer, saltant type probe and wild-type probe, preferably to realize the method based on quantitative fluorescent PCR to detect the mankind
PIK3CD gene c.3061G > A, p.E1021K site mutation, relative to Sanger sequencing and high-flux sequence detection method,
Method based on the quantitative fluorescent PCR realized on the basis of probe provided by the invention and primer is guaranteeing highly sensitive and height
On the basis of specificity, experimental period is shorter, the lower effect of cost.And the fluorescence probe of specificity is relative to common PCR,
False positive, the i.e. appearance of non-specific amplification are reduced again.For the genomic DNA of extraction, the present invention can carry out minimum
The detection of 50ng DNA, mutation rate minimum 10%, and final value≤33 fluorescence curve Ct, illustrate provided by the present invention draw
Object and probe sensitivity with higher.
Detailed description of the invention
Some specific embodiments of the present invention is described in detail by way of example and not limitation with reference to the accompanying drawings hereinafter.
Identical appended drawing reference denotes same or similar part or part in attached drawing.It should be appreciated by those skilled in the art that these
What attached drawing was not necessarily drawn to scale.In attached drawing:
Fig. 1 is that the embodiment of the present invention 1 detects c.3061G > A, and wild type, the E1021K heterozygosis of p.E1021K site mutation are prominent
Modification, E1021K homozygous mutant FAM access diagram；
Fig. 2 is that the embodiment of the present invention 1 detects c.3061G > A, and wild type, the E1021K heterozygosis of p.E1021K site mutation are prominent
Modification, E1021K homozygous mutant CY5 access diagram；
Fig. 3 is that the embodiment of the present invention 3 detects c.3061G > A, the signal of the site the p.E1021K difference channel frequency of mutation FAM
Figure；
Fig. 4 is that the embodiment of the present invention 3 detects c.3061G > A, the signal of the site the p.E1021K difference channel frequency of mutation CY5
Figure.
Specific embodiment
Below technical solution in the embodiment of the present invention describe clear and completely, it is clear that described embodiment
Only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, the common skill in this field
Art personnel every other embodiment obtained without making creative work belongs to the model that the present invention protects
It encloses.
The embodiment of the invention provides one kind for detect mankind PIK3CD gene c.3061G > A, the site p.E1021K is prominent
The probe and primer of change combine, and the primer includes forward primer and reverse primer, and the nucleotide sequence of the forward primer is such as
Shown in SEQ ID NO:1, the nucleotide sequence of the reverse primer is as shown in SEQ ID NO:2；The probe includes saltant type
Probe, the nucleotide sequence of the saltant type probe is as shown in SEQ ID NO:3.
In an alternative embodiment, the molar ratio of the forward primer, reverse primer and the saltant type probe is 1:1:
1。
In an alternative embodiment, the probe further includes wild-type probe, the nucleotide sequence of the wild-type probe
As shown in SEQ ID NO:4.
In an alternative embodiment, 5 ' ends of the saltant type probe and wild-type probe are marked with fluorophor, 3 ' ends
It is marked with quenching group.
In an alternative embodiment, the fluorophor is any one of FAM, CY5, HEX, VIC, ROX.
Wherein the fluorophor and quenching group can be screened based on actual using needs or cost consideration.Specifically
The saltant type probe fluorophor be FAM, quenching group BHQ1；The fluorophor of the wild-type probe is CY5,
Quenching group is BHQ2.Or the fluorophor of the saltant type probe is CY5, quenching group BHQ2；It is described wild
The fluorophor of type probe is FAM, quenching group BHQ1.
Above-mentioned selected two groups of fluorophors and quenching group, realize PIK3CD gene c.3061G > A, p.E1021K
On the basis of site mutation quantitative detection, there is lower use cost.
In an alternative embodiment, the molar ratio of the forward primer and reverse primer is 1:1, the wild-type probe with
The molar ratio of saltant type probe is 1:1, and the molar ratio of the forward primer and the wild-type probe is (1-3): 1.
In a preferred embodiment, the forward primer, reverse primer, wild-type probe and saltant type probe molar ratio
For 1:1:1:1.
For provided by the present invention for detection mankind PIK3CD gene c.3061G > A, p.E1021K site mutation it is prominent
Modification and wild-type probe and primer combination can preferably realize the method detection mankind PIK3CD based on quantitative fluorescent PCR
Gene c.3061G > A, p.E1021K site mutation.The same of the pair of primers of the invention designed is being added when carrying out PCR amplification
When the wild type and saltant type fluorescence probe of the specificity that the present invention designs, probe both ends one fluorophor of label respectively is added
With a quenching group.When probe is complete, the fluorescence signal of fluorophor transmitting is quenched group absorptions；Taq enzyme when PCR amplification
5 ' -3 ' 5 prime excision enzyme activities probe digestion is degraded, separate fluorophor and quenching fluorescence group, thus fluorescence monitoring system
It can receive fluorescence signal, as soon as that is, every amplification DNA chain, has a fluorescent molecule to be formed, realizes the accumulation of fluorescence signal
It is formed with PCR product fully synchronized.If there is mutation, then the primer and probe meeting and target of specificity provided by the invention
DNA profiling combines, and releases fluorescence signal.This fluorescence signal is detected and is converted to readable glimmering by fluorescence quantitative PCR instrument
Light curve is analyzed in conjunction with fluorescence curve, it is known that sample PIK3CD gene c.3061G > A, p.E1021K site mutation
The case where.Substantially increase PIK3CD gene c.3061G > A, p.E1021K site mutation detection sensitivity and specificity.
The embodiment of the present invention also provide it is a kind of for detect mankind PIK3CD gene c.3061G > A, the site p.E1021K is prominent
The kit of change, including it is above-mentioned for detect mankind PIK3CD gene c.3061G > A, the probe of p.E1021K site mutation and
Primer combination.
To those skilled in the art, for above-mentioned offer for detect mankind PIK3CD gene c.3061G > A,
The probe and primer of p.E1021K site mutation combine, and in conjunction with the routine techniques in art technology, can directly acquire to obtain
Preparation for detect mankind PIK3CD gene c.3061G > A, the method for the kit of p.E1021K site mutation.
The embodiment of the present invention also provide it is a kind of it is above-mentioned for detect mankind PIK3CD gene c.3061G > A, p.E1021K
Probe and the primer combination of point preparation for detect mankind PIK3CD gene c.3061G > A, p.E1021K site mutation and prominent
Application in the kit of variability.
The embodiment of the present invention also provide it is a kind of it is above-mentioned for detect mankind PIK3CD gene c.3061G > A, p.E1021K
Probe and the primer combination of point mutation preparation for screening PIK3CD gene c.3061G > A, p.E1021K site mutation is related
PI3K- δ overactivity syndrome product in application.
To those skilled in the art, for above-mentioned offer for detect mankind PIK3CD gene c.3061G > A,
The probe and primer of p.E1021K site mutation combine, and in conjunction with the routine techniques in art technology, can directly acquire to obtain
Preparation for screening PIK3CD gene c.3061G > A, the relevant PI3K- δ overactivity syndrome of p.E1021K site mutation produces
Product.
It is introduced provided by the embodiment of the present invention in detail to become apparent from for detecting mankind PIK3CD gene c.3061G
The probe and primer of > A, p.E1021K site mutation combine and kit, is described below in conjunction with specific embodiment.
Embodiment 1:
One kind for detect mankind PIK3CD gene c.3061G > A, p.E1021K site mutation probe and primer combination,
The primer includes forward primer and reverse primer, and the nucleotide sequence of the forward primer is described as shown in SEQ ID NO:1
The nucleotide sequence of reverse primer is as shown in SEQ ID NO:2；The probe includes saltant type probe and wild-type probe, described
The nucleotide sequence of saltant type probe is as shown in SEQ ID NO:3；The nucleotide sequence of the wild-type probe such as SEQ ID
Shown in NO:4.The saltant type probe and 5 ' the ends label of wild-type probe have, and 3 ' ends label has base
Group.The saltant type fluorescence probe group is FAM, quenching group BHQ1；The wild-type probe fluorophor is CY5, is quenched
The group that goes out is BHQ2.The forward primer, reverse primer, saltant type probe and wild-type probe molar ratio be 1:1:1:1.
Wherein the combination of forward primer, reverse primer and saltant type fluorescence probe can be used to detect PIK3CD gene
C.3061G the mutation of > A, p.E1021K；
And the combination of forward primer, reverse primer and wild-type probe can be used to detect PIK3CD gene c.3061G > A,
The expression of p.E1021K wild type, and not to c.3061G > A, p.E1021K site mutation is detected.
This is because probe is designed in c.3061G > A, p.E1021K site mutation when designing saltant type fluorescence probe
On, it only mutates and just can be carried out normal PCR reaction, release a kind of fluorescence.And wild-type probe design is based on not having
C.3061G when the template of > A, p.E1021K site mutation, only sample are wild type, PCR reaction is normally carried out, and is released another
A kind of fluorescence.PIK3CD can be learned by comparison saltant type probe groups and the different fluorescence curves of wild-type probe group amplification
Gene c.3061G > A, the amount of p.E1021K site mutation and the ratio with unmutated template.Therefore the combination can distinguish open country
Mutation ratio when raw type, heterozygous mutant, homozygous mutant even chimera or generation somatic mutation.
Normal person's wild type site, PIK3CD gene c.3061G > A, p.E1021K heterozygous mutant PI3K- δ overactivity
Syndrome patient, PIK3CD gene c.3061G > A, p.E1021K homozygous mutant PI3K- δ overactivity syndrome patient detection
Result schematic diagram difference is as depicted in figs. 1 and 2.
Embodiment 2:
One kind for detect mankind PIK3CD gene c.3061G > A, the kit of p.E1021K site mutation, including implement
Described in example 1 for detect mankind PIK3CD gene c.3061G > A, p.E1021K site mutation probe and primer combination.Institute
The fluorescence quantitative kit KAPA PROBE FAST qPCR Kits (KAPA of commercially available purchase can directly be selected by stating kit
BIOSYSTEMS, #KK4703).
The kit can also voluntarily be prepared, such as the kit may also include and routinely add in the kit of this field
Component, further comprise but be not limited to following component: primer buffer, probe buffer, PCR buffer, Taq
Enzyme, archaeal dna polymerase, Mg2+, dNTP etc..Above content that those skilled in the art are provided based on the embodiment of the present invention 2 and existing
Technology can be directly obtained corresponding kit.
Embodiment 3:
Provided by embodiment 2 for detect mankind PIK3CD gene c.3061G > A, the reagent of p.E1021K site mutation
The test of box effect detection:
The genomic DNA and fluorescence quantitative kit KAPA PROBE mentioned using peripheral blood, hair, buccal mucosa
FAST qPCR Kits (KAPA BIOSYSTEMS, #KK4703) is tested, and experimental system size is recommended according to kit
It is determined as 20 μ l.
In the case where experimental situation and other conditions all the same, draw to the above-mentioned forward primer in experimental system and reversely
Test is optimized in object concentration, and the final final concentration for determining 500nM can reach best detection effect.
In the case where experimental situation and other conditions all the same, the fluorescence probe concentration in experimental system is carried out excellent
Change test, the final final concentration for determining 500nM can reach best detection effect.
Finally determining optimal reaction system is as shown in table 1.
Reaction condition is tested after repeatedly optimizing, is ultimately determined to: 95 DEG C of 5min；95 DEG C of 15s, 60 DEG C of 1min, 45
A circulation；40℃30s.
1 optimal reaction system of table
| Composition | Volume (μ L) |
| 2×KAPA Probe Mix | 10μl |
| DNA profiling | 50ng |
| Probe (10 μM) | 1μl |
| Forward primer (10 μM) | 1μl |
| Reverse primer (10 μM) | 1μl |
| Water | Complement to 20 μ l |
| Total volume | 20μl |
Design synthesis containing PIK3CD gene c.3061G > A, the plasmid (frequency of mutation of p.E1021K site mutation
100%), and completely wild type PIK3CD plasmid (frequency of mutation 0%).Configure the positive sample of the different frequencies of mutation
(0%, 10%, 20%, 30%, 50%) carries out Monitoring lower-cut experiment.
Experimental result is as shown in Fig. 3 and Fig. 4 it is found that kit of the present invention can carry out minimum 50ngDNA, mutation
The detection of rate minimum 10%, and final value≤33 fluorescence curve Ct.Show primer and probe combination provided by the present invention
And kit has good sensitivity.
Sequence table
<110>Chinese Academy of Medical Sciences Beijing Union Medical College Hospital
<120>it is used to detect the probe of PIK3CD gene E1021K site mutation and primer combines and kit
<130> CC18K10282CCN
<141> 2018-12-24
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 18
<212> DNA
<213> Artificial Sequence
<400> 1
gggaaaacag aggaggag 18
<210> 2
<211> 18
<212> DNA
<213> Artificial Sequence
<400> 2
ccctttagga ccaatgtg 18
<210> 3
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 3
acggagggct ttgttaaact tcac 24
<210> 4
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 4
acggagggct tcgttaaact tcac 24
Claims (8)
1. one kind for detect mankind PIK3CD gene c.3061G > A, p.E1021K site mutation probe and primer combination,
Be characterized in that: the primer includes forward primer and reverse primer, the nucleotide sequence of the forward primer such as SEQ ID NO:1
Shown, the nucleotide sequence of the reverse primer is as shown in SEQ ID NO:2；The probe includes saltant type probe, described prominent
The nucleotide sequence of modification probe is as shown in SEQ ID NO:3.
2. it is according to claim 1 for detect mankind PIK3CD gene c.3061G > A, the spy of p.E1021K site mutation
Needle and primer combination, it is characterised in that: the molar ratio of the forward primer, reverse primer and the saltant type probe is 1:1:1.
3. it is according to claim 1 for detect mankind PIK3CD gene c.3061G > A, the spy of p.E1021K site mutation
Needle and primer combination, it is characterised in that: the probe further includes wild-type probe, and the nucleotide sequence of the wild-type probe is such as
Shown in SEQ ID NO:4.
4. it is according to claim 3 for detect mankind PIK3CD gene c.3061G > A, the spy of p.E1021K site mutation
Needle and primer combination, it is characterised in that: the saltant type probe and 5 ' the ends label of wild-type probe have, 3 ' ends
Label has.
5. it is according to claim 4 for detect mankind PIK3CD gene c.3061G > A, the spy of p.E1021K site mutation
Needle and primer combination, it is characterised in that: the molar ratio of the forward primer and reverse primer be 1:1, the wild-type probe with
The molar ratio of saltant type probe is 1:1, and the molar ratio of the forward primer and the wild-type probe is (1-3): 1.
6. one kind for detect mankind PIK3CD gene c.3061G > A, the kit of p.E1021K site mutation, feature exists
In: it is described in any item for detecting the site mankind PIK3CD gene C .3061G > A, P.E1021K including Claims 1 to 5
Probe and primer combination.
7. a kind of Claims 1 to 5 it is described in any item for detect mankind PIK3CD gene c.3061G > A, p.E1021K
Probe and the primer combination of point preparation for detect mankind PIK3CD gene c.3061G > A, p.E1021K site mutation and prominent
Application in the kit of variability.
8. a kind of Claims 1 to 5 it is described in any item for detect mankind PIK3CD gene c.3061G > A, p.E1021K
Probe and the primer combination of point mutation preparation for screening PIK3CD gene c.3061G > A, p.E1021K site mutation is related
Application in the product of PI3K δ overactivity syndrome.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811584627.0A CN109337976A (en) | 2018-12-24 | 2018-12-24 | For detect PIK3CD gene E1021K site mutation probe and primer combination and kit |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811584627.0A CN109337976A (en) | 2018-12-24 | 2018-12-24 | For detect PIK3CD gene E1021K site mutation probe and primer combination and kit |
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Family
ID=65297068
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| WO2013043878A2 (en) * | 2011-09-20 | 2013-03-28 | The George Washington University | Alternative splicing variants of genes associated with prostate cancer risk and survival |
| CN105339507A (en) * | 2013-02-21 | 2016-02-17 | 托马生物科学公司 | Methods, compositions, and kits for nucleic acid analysis |
| CN105658218A (en) * | 2013-10-17 | 2016-06-08 | 葛兰素史克知识产权开发有限公司 | Pi3k inhibitor for treatment of respiratory disease |
| CN107922973A (en) * | 2015-07-07 | 2018-04-17 | 远见基因组系统公司 | Method and system for the modification detection based on sequencing |
-
2018
- 2018-12-24 CN CN201811584627.0A patent/CN109337976A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013043878A2 (en) * | 2011-09-20 | 2013-03-28 | The George Washington University | Alternative splicing variants of genes associated with prostate cancer risk and survival |
| CN105339507A (en) * | 2013-02-21 | 2016-02-17 | 托马生物科学公司 | Methods, compositions, and kits for nucleic acid analysis |
| CN105658218A (en) * | 2013-10-17 | 2016-06-08 | 葛兰素史克知识产权开发有限公司 | Pi3k inhibitor for treatment of respiratory disease |
| CN107922973A (en) * | 2015-07-07 | 2018-04-17 | 远见基因组系统公司 | Method and system for the modification detection based on sequencing |
Non-Patent Citations (5)
| Title |
|---|
| IVAN ANGULO等: "Phosphoinositide 3-Kinase d Gene Mutation Predisposes to Respiratory Infection and Airway Damage", 《SCIENCE》 * |
| NEJEN: "ss836318712", 《DBSNP DATABASE》 * |
| SIEMPELKAMP BD等: "Homo sapiens phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit delta (PIK3CD), transcript variant 1, mRNA", 《GENBANK DATABASE》 * |
| 唐文静等: "PIK3CD基因突变致PI3Kδ过度活化综合征临床及免疫学特点分析", 《中华儿科杂志》 * |
| 林果为等: "《现代临床血液病学》", 31 August 2013, 复旦大学出版社 * |
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Application publication date: 20190215 |