CN106609300A - Coronary artery disease risk assessment kit and risk assessment method - Google Patents
Coronary artery disease risk assessment kit and risk assessment method Download PDFInfo
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Abstract
The embodiment of the invention discloses a coronary artery disease risk assessment kit and a risk assessment method, and is applied in the field of disease risk assessment. The risk assessment method comprises the steps: extracting peripheral blood as a sample; extracting peripheral blood RNA by a peripheral blood RNA extraction kit; carrying out reverse transcription on the peripheral blood RNA, and constructing a cDNA library; carrying out high throughput sequencing of the cDNA library to obtain a sequencing sequence; carrying out gene comparison; calculating the gene expression quantity; calculating a score of coronary artery stenosis; and assessing whether the risk of suffering from coronary artery disease exists. The coronary artery disease risk assessment method provide by the invention has the advantages of no invasion and high accuracy rate.
Description
Technical field
The present invention relates to disease risks evaluation areas, more particularly to a kind of assessment examination of coronary artery disease risk
Agent box and methods of risk assessment.
Background technology
The M ＆ M that today's society, coronary heart disease (CAD) and myocardial infarction (MI) cause is
The main health burden in the whole world.At present, the clinical factor of coronary heart disease has a lot, such as sex, age, breast
Pain type, smoking history, diabetes etc..Additionally, atherosclerotic generation is closely related with inflammation.
Some inflammatory cells are gathered in the periendothelial of damage with the factor, promote atherosclerotic formation and easy
Damage the rupture of speckle.Due to blood circulation, these changes are indirectly shown in peripheral blood, quantitative determination
The expression of related gene can reflect the order of severity of coronary heart disease in peripheral blood.
In prior art, clinically the conventional detection method of coronary heart disease has：Electrocardiogram, ultrasoundcardiogram,
Stress test, CT coronarographies, coronarography (CAG) etc..Electrocardiogram, ultrasonic cardiography
Figure, stress test is limited due to Sensitivity and Specificity, therefore, its negative inspection result can not be arranged
Except coronary heart disease.So coronarography (CAG) remains " goldstandard " of diagnosis of coronary heart disease, but by
In technology is relative complex, it is invasive, costly, and there is certain risk, cause many patients to be difficult to connect
Receive, in addition according to statistics, intervened only less than 1/3 in the patient for carrying out coronarography (CAG)
Treatment, most patient carries out coronarography (CAG) and is intended merely to determine diagnosis.And CT
Coronarography (CTA) possesses that result is accurate, wound is little, the advantage of expense relative moderate, compared with
Well for clinic.But due to the negative predictive value height of CT coronarographies (CTA), therefore
The negative diagnostic value of CT coronarographies (CTA) is larger, and positive diagnosis value is limited, especially
It is the judgement for stenosis, so CT coronarographies (CTA) are mainly used in screening.
Nearly 2 years, high throughput sequencing technologies had important breakthrough, the sequencing skill of a new generation in biological technical field
Traditional Sanger sequencings efficiency is improve hundreds times by art, while price is also greatly reduced.High pass is measured
The appearance of sequence technology causes many extremely promising biological medicine applications to be possibly realized.And transcript profile is referred to
The summation of particular organization or cell all RNA of institute's transcriptional expression under a certain environment or physiological condition,
That generally interested is the mRNA of encoding proteins, so high throughput sequencing technologies are combined into transcription group
Research gene expression and regulation provides important means and method.
The content of the invention
It is an object of the invention to the risk assessment for solving some coronary artery diseases of the prior art is used
Method there is a problem that costly, risk is big, although also some appraisal procedures to there is expense low,
But the limited problem of value is judged, so coronary artery disease risk appraisal procedure of the prior art is not
Can fast and safely and low cost assessment patient's coronary thrombosiss risk.
In view of this, the present invention provides a kind of methods of risk assessment of coronary artery disease, it may include：
Anticoagulant tube containing RNase inhibitor extracts peripheral blood as sample；
Peripheral blood RNA extracts kits extract the peripheral blood RNA in the sample；
Reverse transcription and construction cDNA text are carried out to the peripheral blood RNA by RNA reverse transcription reagent box
Storehouse；
High-flux sequence is carried out to the cDNA library and obtains sequencing sequence；
The sequencing sequence and human rna genome database are carried out into sequence alignment uniquely to be compared
The measure sequence of genes of interest；
By 100- (unique measure sequence number/unique surveys for comparing upper genes of interest for comparing upper term single gene
Sequencing row sum) * 10000 obtain gene expression amount formula；
Each gene is substituted into into the gene expression amount that the gene expression amount formula is calculated each gene；
Coronary stenosis scoring formula is substituted into by the gene expression amount by each gene and calculates coronary stenosis scoring；
Judge size of the coronary stenosis scoring with 0, if coronary stenosis scoring is less than 0, do not have
The risk of coronary artery disease.
Present invention also offers a kind of risk assessment reagent kit of coronary artery disease, it may include blood taking tube,
RNA extracts kits and RNA reverse transcription reagent box, the blood taking tube is used to take peripheral blood sample,
The RNA extracts kits are used to extract outer peripheral blood in the peripheral blood sample taken from the blood taking tube
RNA, the RNA reverse transcription reagent box is used to carry out reverse transcription to the peripheral blood RNA and build
CDNA library.
As can be seen from the above technical solutions, the present invention has advantages below：
In the present invention, high pass sequencing is carried out by the RNA in human peripheral blood sample, and compare gene, counted
Coronary stenosis scoring is calculated, by the risk of scoring assessment patient's coronary thrombosiss.The method is without the need for suffering from
Person carries out traumatic operation, possesses noninvasive advantage.Compared to CT coronary angiographies, what the present invention was provided
Accurate, noninvasive advantage that method has, compared to electrocardiogram and cardiac stress test accuracy rate also significantly
Improve.Therefore, the present invention fast and safely have evaluated by the change of rna level in detection patient's body
The risk of patient's coronary thrombosiss, with good clinical meaning and practical value.
Description of the drawings
Fig. 1 is a kind of methods of risk assessment flow chart of coronary artery disease of the present invention；
Fig. 2 is a kind of risk assessment reagent kit structure chart of coronary artery disease of the present invention.
Specific embodiment
A kind of methods of risk assessment of coronary artery disease is embodiments provided, be can solve the problem that existing
It is big that the method that the risk assessment of some coronary artery diseases in technology is used has costly, risk
Problem, although also some appraisal procedures have that expense is low, judges the limited problem of value.
In order that those skilled in the art more fully understand the present invention program, below in conjunction with of the invention real
The accompanying drawing in example is applied, the technical scheme in the embodiment of the present invention is clearly and completely described, it is clear that
Described embodiment is only the embodiment of a present invention part, rather than the embodiment of whole.It is based on
Embodiment in the present invention, those of ordinary skill in the art are obtained under the premise of creative work is not made
The every other embodiment for obtaining, should all belong to the scope of protection of the invention.
Fig. 1 is referred to, a kind of methods of risk assessment flow chart of the coronary artery disease provided for the present invention,
As seen from Figure 1, methods described includes：
S101, the anticoagulant tube containing RNase inhibitor extract peripheral blood as sample；
S102, peripheral blood RNA extracts kits extract the peripheral blood RNA in the sample；
S103, reverse transcription is carried out to the peripheral blood RNA by RNA reverse transcription reagent box and is built
CDNA library；
S104, high-flux sequence is carried out to the cDNA library obtain sequencing sequence；
S105, the sequencing sequence and human rna genome database are carried out into sequence alignment obtain uniquely
The measure sequence of genes of interest in comparison；
S106, by 100-, (unique measure sequence number/unique for comparing upper term single gene compares upper purpose base
The measure sequence sum of cause) * 10000 obtain gene expression amount formula；
S107, the gene table that each described gene expression amount formula of gene substitution is calculated each gene
Up to amount；
S108, the formula calculating arteria coronaria that scored by the gene expression amount substitution coronary stenosis by each gene are narrow
Narrow scoring；
S109, judge size of the coronary stenosis scoring with 0, if coronary stenosis scoring is less than 0,
The then risk without coronary artery disease.
Specifically, the method that the present invention is provided extracts trouble using by the anticoagulant tube containing RNase inhibitor
The peripheral blood of person.After peripheral blood in patients is obtained, peripheral blood RNA extracts kits can be supported the use and entered
Row RNA is extracted, and obtains the peripheral blood RNA in the sample.Using commercially available RNA Reverse Transcriptions
RNA is carried out reverse transcription by box, and builds acquisition cDNA library.By high-flux sequence platform to building
CDNA library carry out sequencing and obtain sequencing sequence, then by the sequencing sequence and related data base
Carry out sequence alignment and obtain unique measure sequence for comparing upper genes of interest.Coronary stenosis scoring is finally calculated,
The risk of patient's coronary thrombosiss is assessed by judging size of the coronary stenosis scoring with 0.
A kind of methods of risk assessment of coronary artery disease that the present embodiment is provided is by human peripheral blood sample
RNA carry out high pass sequencing, and compare gene, coronary stenosis scoring is calculated, by scoring assessment patient
The risk of coronary thrombosiss.The method possesses noninvasive excellent without the need for carrying out traumatic operation to patient
Point.Compared to CT coronary angiographies, the method that the present invention is provided has accurate, noninvasive advantage, compared to
The accuracy rate of electrocardiogram and cardiac stress test is also greatly improved.Therefore, the present invention is by detecting patient's body
The change of interior rna level, fast and safely have evaluated the risk of patient's coronary thrombosiss, with good
Clinical meaning and practical value.
A kind of beneficial effect of the ECG detection device for the ease of providing the present invention has one more intuitively
Understand, present invention also offers embodiment 2, with reference to a kind of methods of risk assessments of coronary artery disease of Fig. 1
Flow process is described in detail.
S101, the anticoagulant tube containing RNase inhibitor extract peripheral blood as sample；
Specifically, peripheral blood in patients is extracted by the anticoagulant tube containing RNase inhibitor, can be in room temperature
(16-25 DEG C) preserves 5 days, and 2-8 DEG C preserves 7 days.It should be noted that common anticoagulant tube can also be used
Take out without peripheral blood in patients, the peripheral blood sample that this mode is obtained then is needed in 4 DEG C of preservations, and must be 4
Detected within hour.
Also, it should be noted that the preservation of the peripheral blood sample can also be replaced using blood card.
S102, peripheral blood RNA extracts kits extract the peripheral blood RNA in the sample；
Specifically, obtain after peripheral blood in patients, can support the use peripheral blood RNA extracts kits is carried out
RNA is extracted, and extracted amount at least 1ng, extracting method can be found in kit specification.
S103, reverse transcription is carried out to the peripheral blood RNA by RNA reverse transcription reagent box and is built
CDNA library；
Specifically, the construction cDNA library may include：To obtaining after the peripheral blood RNA reverse transcriptions
CDNA amplification enrichment obtain full-length cDNA；The full-length cDNA is broken into into fragment；To described
Fragment carries out screening the cDNA fragments for obtaining 200bp scopes；CDNA pieces to the 200bp scopes
Duan Jinhang repairs and adds joint in flat end, then enters performing PCR amplification, obtains the cDNA library.
It should be noted that RNA reverse transcription reagent box can be the test kit of market sale, concrete grammar
May refer to cDNA transcription description.
S104, high-flux sequence is carried out to the cDNA library obtain sequencing sequence；
Specifically, after the cDNA library is obtained, sequencing sequence is obtained by high-flux sequence.It is excellent
Choosing, high-flux sequence platform is any one in Illumina microarray datasets or Ion torrent microarray datasets
Kind.Preferably, can be as carrying out joint, going low quality, go to sequence obtained by Illumina sequencings
Pollution etc. is processed, and finally obtains clean mRNA molecular sequences.
S105, the sequencing sequence and human rna genome database are carried out into sequence alignment obtain uniquely
The measure sequence of genes of interest in comparison；
Specifically, by the clean mRNA molecular sequences of above-mentioned acquisition and human rna genomic data
Storehouse carries out sequence alignment, it is possible to obtain unique measure sequence for comparing upper genes of interest.Preferably, it is described
Human rna genome database includes repeatmarker data bases, Genbank data bases, Rfam (10.1)
One or more in data base, UCSC data bases and piRNA data bases.Preferably, can pass through
Using the softwares of Microsoft SOAP Toolkit 2.0 by clean mRNA molecular sequences and human rna base
Because group data base carries out sequence alignment.
It should be noted that in unique comparison genes of interest measure sequence include AF28, AQP9,
CASP5、CD3D、CD79B、CLEC4E、HNRNPF、IL18RAP、IL8RB、KCNE3、
KLRC4、NCF4、RPL28、S100A12、S100A8、SLAMF7、SPIB、TFCP2、TLR4、
TMC8、TNFAIP6、TNFRSF10C。
S106, by 100-, (unique measure sequence number/unique for comparing upper term single gene compares upper purpose base
The measure sequence sum of cause) * 10000 obtain gene expression amount formula；
Specifically, by 100-, (unique measure sequence number/unique for comparing upper term single gene compares upper purpose
The measure sequence sum of gene) * 10000 describing gene expression amount.It is understood that unique compare
Upper genes of interest measure sequence sum be AF28 noted earlier, AQP9, CASP5, CD3D,
CD79B、CLEC4E、HNRNPF、IL18RAP、IL8RB、KCNE3、KLRC4、NCF4、
RPL28、S100A12、S100A8、SLAMF7、SPIB、TFCP2、TLR4、TMC8、TNFAIP6、
The measure sequence sum of TNFRSF10C this 22 genes of interest.
S107, the gene table that each described gene expression amount formula of gene substitution is calculated each gene
Up to amount；
Specifically, each comparison gene can be updated in the gene expression amount formula, obtains every
The expression of individual gene.
S108, the formula calculating arteria coronaria that scored by the gene expression amount substitution coronary stenosis by each gene are narrow
Narrow scoring；
Specifically, the expression of each gene obtained above is updated to into the coronary stenosis scoring formula
In, it is calculated coronary stenosis scoring.
Preferably, the coronary stenosis scoring formula can include
1) Norm1=RPL28=22.13 is defined；
2) Norm2=(0.5*HNRPF+0.5*TFCP2) is defined；
3) NKup=(0.5*SLAMF7+0.5*KLRC4) 4 is defined)；
4) Tcell=(0.5*CD3D+0.5*TMC8) is defined；
5) Bcell=(2/3*CD79B+1/3*SPIB) is defined；
6) Neut=(0.5*AQP9+0.5*NCF4) is defined；
7) Nup=(1/3*CASP5+1/3*IL18RAP+1/3*TNFAIP6) is defined；
8) Ndown=(0.25*IL8RB+0.25*TNFRSF10C+0.25*TLR4+ are defined
0.25*KCNE3)；
9) SCA1=(1/3*S100A12+1/3*CLEC4E+1/3*S100A8) is defined；
10) AF2=AF289562 is defined；
11) female SEX=0, male SEX=1 are defined；
12) Intercept female is defined：INTERCEPT=1.821+0.123* (age -60), as a result for it is negative then
It is set to 0；
Raw Score=INTERCEPT-0.755* (Nup-Ndown) -0.308*SEX* (SCA1-Norm1) -
0.548*(1-SEX)*(SCA1-Neut)-0.406*(NKup-Tcell)-0.137*(Bcell-Tcell)-0.482*SE
X-0.246(AF2-Norm2)；
Final score Final Score=RawScore*40/4.52.
It is understood that each formula definition in coronary stenosis scoring formula as Norm1,
The contents such as Norm2, are intended only as a variable, convenient to calculate coronary stenosis scoring, without physical meaning.
S109, judge size of the coronary stenosis scoring with 0, if coronary stenosis scoring is less than 0,
The then risk without coronary artery disease.
Specifically, the final score that coronary stenosis obtained above score is compared with 0, by sentencing
Size of the disconnected coronary stenosis scoring with 0 is assessing the risk of the coronary artery disease of patient.
Wherein, if coronary stenosis scoring is less than 0, assessing patient does not have the illness of coronary artery disease
Risk, i.e. result judge：Final Score<0 patient is normal.
For the clearer assessment for understanding a kind of coronary artery disease risk appraisal procedure that the present invention is provided
Process, additionally provides embodiment 3, the coronary artery disease risk is elaborated by taking actual patient as an example and is commented
Estimate the course of work of method.
(1) preferred, female patient at 51 years old age, before Coronary Artery in patients radiography, extracts patient
Peripheral blood 3ml, collects in TempusTMIn Blood RNA Tube, preserve 3 days in 4 DEG C.
(2) it is preferred, use TempusTMSpin RNA Isolation Kit isolate and purify RNA, specifically
Operation can be found in kit specification, finally obtain 760ngRNA.
(3) it is preferred, using SMARTer PCR cDNA synthesis kit test kits to said extracted
The 760ng RNA for obtaining carry out reverse transcription and corresponding amplification enrichment, the visible reverse transcription explanation of concrete operations
Book.
(4) it is preferred, using the broken instrument of Covaris ultrasound wave DNA, according to instrument description by richness
The full-length cDNA of collection is broken into fragment.
(5) it is preferred, useXP (Nucleic acid purification kits) is carried out
Fragment is screened, and selects the cDNA fragments of 200bp scopes.
(6) it is preferred, useUltraTMDNA Library Prep Kit for Illumina (are used
In the super DNA library test kit of Illumina) carry out flat end reparation, plus joint.
(7) it is preferred, useMultiplex Oligos for(it is used for Illumina
Multisample adapter-primer test kit) enter performing PCR amplification, obtain 340ng cDNA libraries.
(8) it is preferred, high pass is carried out to sequencing sequence using the microarray datasets of Illumina NextSeq 500
Sequence is measured, as a result as shown in table 1：
Table 1：Sequencing master data analysis
(9) using the softwares of Microsoft SOAP Toolkit 2.0 by clean mRNA molecular sequences and people
Class rna gene group data base carries out sequence alignment, then each gene is updated to into gene expression amount formula
In, that is, substituting into the results of 100- percentage ratios * 10000, to bring formula result of calculation into as shown in table 2：
Table 2：Sequencing result distribution table
It should be noted that wherein percentage ratio is unique measure sequence number/unique ratio for comparing upper term single gene
Measure sequence sum to upper genes of interest.
(10) expression of each gene is updated in coronary stenosis scoring formula, finally calculates score
For -14.8, the score is clearly less than 0, then finally assessing 51 years old female patient does not have coronary artery disease
The risk of disease.
A kind of coronary artery disease risk appraisal procedure provided in an embodiment of the present invention to patient without the need for creating
The operation of wound property, with noninvasive advantage, while by clinical verification, negative accuracy rate up to 96%,
Sensitivity 89%, specificity 52% is particularly suited for the exclusion of suspected patient, makes compared to CT arteria coronaria
Shadow possesses accurately, noninvasive advantage, greatly improves compared to the accuracy rate of electrocardiogram and cardiac stress test.
In addition according to statistics, more than 60% patient for receiving coronary angiography does not have coronary heart disease in fact, therefore for
The exclusion of negative patient seems particularly necessary, and the embodiment of the present invention is by rna level in detection patient's body
Change, fast and safely assesses the risk of patient's coronary thrombosiss, with good clinical meaning and reality
With value.
In addition, the present invention further correspondingly provides a kind of risk assessment reagent kit of coronary artery disease, specifically
Referring to the drawings shown in 2, blood taking tube, RNA extracts kits and RNA reverse transcription reagent box can be included,
The blood taking tube is used to take peripheral blood sample, the RNA extracts kits to be used for from the blood taking tube
Outer peripheral blood RNA is extracted in the peripheral blood sample taken, the RNA reverse transcription reagent box is used for institute
Stating peripheral blood RNA carries out reverse transcription and construction cDNA library.
Term " first ", " second " in description and claims of this specification and above-mentioned accompanying drawing, "
Three " (if present) such as " 4th " is the object for distinguishing similar, without specific for describing
Order or precedence.It should be appreciated that the data for so using can be exchanged in the appropriate case, so as to this
In describe embodiment can with addition to the content for illustrating here or describe order enforcement.Additionally,
Term " comprising " and " having " and their any deformation, it is intended that covering is non-exclusive to be included,
For example, process, method, system, product or the equipment for containing series of steps or unit is not necessarily limited to
Those steps clearly listed or unit, but may include clearly not list or for these mistakes
Other intrinsic steps of journey, method, product or equipment or unit.
The above, above example only to illustrate technical scheme, rather than a limitation；
Although being described in detail to the present invention with reference to the foregoing embodiments, one of ordinary skill in the art should
Work as understanding：It still can modify to the technical scheme described in foregoing embodiments, or to it
Middle some technical characteristics carry out equivalent；And these modifications or replacement, do not make appropriate technical solution
Essence depart from various embodiments of the present invention technical scheme spirit and scope.
Claims (9)
1. a kind of methods of risk assessment of coronary artery disease, it is characterised in that methods described includes：
Anticoagulant tube containing RNase inhibitor extracts peripheral blood as sample；
Peripheral blood RNA extracts kits extract the peripheral blood RNA in the sample；
Reverse transcription and construction cDNA text are carried out to the peripheral blood RNA by RNA reverse transcription reagent box
Storehouse；
High-flux sequence is carried out to the cDNA library and obtains sequencing sequence；
The sequencing sequence and human rna genome database are carried out into sequence alignment uniquely to be compared
The measure sequence of genes of interest；
By 100- (unique measure sequence number/unique surveys for comparing upper genes of interest for comparing upper term single gene
Sequencing row sum) * 10000 obtain gene expression amount formula；
Each gene is substituted into into the gene expression amount that the gene expression amount formula is calculated each gene；
Coronary stenosis scoring formula is substituted into by the gene expression amount by each gene and calculates coronary stenosis scoring；
Judge size of the coronary stenosis scoring with 0, if coronary stenosis scoring is less than 0, do not have
The risk of coronary artery disease.
2. method according to claim 1, it is characterised in that the construction cDNA library includes：
CDNA amplification enrichments to obtaining after the peripheral blood RNA reverse transcriptions obtain full-length cDNA；
The full-length cDNA is broken into into fragment；
The fragment is carried out to screen the cDNA fragments for obtaining 200bp scopes；
Flat end is carried out to the cDNA fragments of the 200bp scopes and repairs and add joint, then enter performing PCR
Amplification, obtains the cDNA library.
3. method according to claim 1, it is characterised in that the human rna genome number
Include repeatmarker data bases, Genbank data bases, Rfam (10.1) data base, UCSC according to storehouse
One or more in data base and piRNA data bases.
4. method according to claim 1, it is characterised in that genes of interest in unique comparison
Measure sequence include AF28, AQP9, CASP5, CD3D, CD79B, CLEC4E, HNRNPF,
IL18RAP、IL8RB、KCNE3、KLRC4、NCF4、RPL28、S100A12、S100A8、
SLAMF7、SPIB、TFCP2、TLR4、TMC8、TNFAIP6、TNFRSF10C。
5. method according to claim 1, it is characterised in that the coronary stenosis score formula bag
Include：
1) Norm1=RPL28=22.13 is defined；
2) Norm2=(0.5*HNRPF+0.5*TFCP2) is defined；
3) NKup=(0.5*SLAMF7+0.5*KLRC4) 4 is defined)；
4) Tcell=(0.5*CD3D+0.5*TMC8) is defined；
5) Bcell=(2/3*CD79B+1/3*SPIB) is defined；
6) Neut=(0.5*AQP9+0.5*NCF4) is defined；
7) Nup=(1/3*CASP5+1/3*IL18RAP+1/3*TNFAIP6) is defined；
8) Ndown=(0.25*IL8RB+0.25*TNFRSF10C+0.25*TLR4+ are defined
0.25*KCNE3)；
9) SCA1=(1/3*S100A12+1/3*CLEC4E+1/3*S100A8) is defined；
10) AF2=AF289562 is defined；
11) female SEX=0, male SEX=1 are defined；
12) Intercept female is defined：INTERCEPT=1.821+0.123* (age -60), as a result for it is negative then
It is set to 0；
Raw Score=INTERCEPT-0.755* (Nup-Ndown) -0.308*SEX* (SCA1-Norm1) -
0.548*(1-SEX)*(SCA1-Neut)-0.406*(NKup-Tcell)-0.137*(Bcell-Tcell)-0.482*SE
X-0.246(AF2-Norm2)；
Final Score=RawScore*40/4.52.
6. method according to claim 1, it is characterised in that the peripheral blood sample is protected in room temperature
Deposit less than 5 days.
7. method according to claim 6, it is characterised in that the extraction of the peripheral blood sample makes
During with common anticoagulant tube, cryopreservation is less than 4 hours.
8. method according to claim 1, it is characterised in that the high-flux sequence platform is
Any one in Illumina microarray datasets or Ion torrent microarray datasets.
9. a kind of risk assessment reagent kit of coronary artery disease, it is characterised in that the test kit includes：
Blood taking tube, RNA extracts kits and RNA reverse transcription reagent box, the blood taking tube is used to take periphery
Blood sample, the RNA extracts kits are used to be extracted in the peripheral blood sample taken from the blood taking tube
Outer peripheral blood RNA, the RNA reverse transcription reagent box is used to carry out reverse transcription to the peripheral blood RNA
And construction cDNA library.
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| CN107164378A (en) * | 2017-06-20 | 2017-09-15 | 浙江大学 | Specificity suppresses siRNA and its recombinant vector and the application of SPIB gene expressions |
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| CN109852689A (en) * | 2019-04-03 | 2019-06-07 | 上海交通大学医学院附属第九人民医院 | The relevant biomarker of one group of vascular malformation and coherent detection kit |
| CN111354464A (en) * | 2018-12-24 | 2020-06-30 | 深圳先进技术研究院 | CAD prediction model establishing method and device and electronic equipment |
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| CN104774959A (en) * | 2015-04-27 | 2015-07-15 | 刘志东 | Kit for detecting blood circulation tumor cells |
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| CN104419753A (en) * | 2013-08-29 | 2015-03-18 | 公安部物证鉴定中心 | Method and system for identifying histologic origin of body fluids of Chinese population from gene level |
| CN104774959A (en) * | 2015-04-27 | 2015-07-15 | 刘志东 | Kit for detecting blood circulation tumor cells |
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| CN107164378A (en) * | 2017-06-20 | 2017-09-15 | 浙江大学 | Specificity suppresses siRNA and its recombinant vector and the application of SPIB gene expressions |
| CN107164378B (en) * | 2017-06-20 | 2019-12-31 | 浙江大学 | siRNA for specifically inhibiting SPIB gene expression and recombinant vector and application thereof |
| CN111354464A (en) * | 2018-12-24 | 2020-06-30 | 深圳先进技术研究院 | CAD prediction model establishing method and device and electronic equipment |
| CN111354464B (en) * | 2018-12-24 | 2024-05-17 | 深圳先进技术研究院 | CAD prediction model establishment method and device and electronic equipment |
| CN109852689A (en) * | 2019-04-03 | 2019-06-07 | 上海交通大学医学院附属第九人民医院 | The relevant biomarker of one group of vascular malformation and coherent detection kit |
| CN109852689B (en) * | 2019-04-03 | 2022-02-18 | 上海交通大学医学院附属第九人民医院 | Group of vascular malformation related biomarkers and related detection kit |
| CN113317879A (en) * | 2021-05-28 | 2021-08-31 | 郑州大学第一附属医院 | Kit for predicting TIPS postoperative stent restenosis of liver cirrhosis patient |
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