CN110317877A - Application of the unstable variation of one group chromosome in preparation diagnosis bladder transitional cell carcinoma, the reagent or kit of assessing prognosis - Google Patents
Application of the unstable variation of one group chromosome in preparation diagnosis bladder transitional cell carcinoma, the reagent or kit of assessing prognosis Download PDFInfo
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Abstract
The present invention relates to biotechnologys and area of medical diagnostics, application of the specifically group chromosome unstable region in the reagent or kit of preparation diagnosis bladder transitional cell carcinoma, it includes following 10 that the chromosome instability, which determines region: 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q, 17p, 17q.The bladder transitional cell carcinoma patient of present invention collection clinical definite, the sample of Healthy People and non-bladder transitional cell carcinoma Disease, pass through genome sequencing comparison of tumor patient and the chromosomal abnormalities that compare, illustrate chromosome instability distribution characteristics specific to bladder transitional cell carcinoma, it discloses 10 common chromosome instabilities of bladder transitional cell carcinoma patient and determines region, total carrying rate is up to 93.4%, this diagnosis to clinically bladder transitional cell carcinoma, treatment, monitoring meter prognosis evaluation has a very big significance, scientific basis is provided to be early diagnosed and being formulated individualized treatment scheme in next step.
Description
Technical field
The present invention relates to biotechnologys and medical diagnostic techniqu field, specifically, being that one group of bladder transitional cell carcinoma is relevant
Chromosome instability variation and the variation of these unstability are in preparation diagnosis bladder transitional cell carcinoma, the reagent for assessing prognosis or examination
Application in agent box.
Background technique
Bladder transitional cell carcinoma (urotheilial cancer, UC) refers to generation on mucous membrane of urinary bladder, ureter epithelium, renal plevis
Malignant tumour.It is one of the most common malignant tumour of urinary system and the big kinds of tumor of whole body ten.It is raw to occupy China's uropoiesis
Grow is first of tumor invasion.WHO " urinary system and genital orgnas,male's oncological pathology and science of heredity " middle urine in 2004
In the system tumor histologic classification of road the histological type of bladder transitional cell carcinoma include Urothelial Carcinoma of Bladder, squamous cell carcinoma of bladder,
Adenocarcinoma of bladder, other rare also bladder clear cell carcinoma, bladder small cell carcinoma, bladder class cancers.One of the most common is wing
Guang bladder transitional cell carcinoma accounts for about 90% or more of bladder transitional cell carcinoma patient summary, and usually said bladder transitional cell carcinoma just refers to bladder
Bladder transitional cell carcinoma.The cause of disease of bladder transitional cell carcinoma is complicated, existing inherence inherent cause, and has external environmental factor, more clear
Two greatly cure the disease risk factors be smoking and Exposed aromatic amine chemical substance.The bladder transitional cell carcinoma of 30%-50% is by inhaling
Cigarette causes, and smoking can make bladder transitional cell carcinoma level of significance increase 2-6 times.Another important cure the disease risk factor and a series of occupations or duty
Industry contact is related, and aniline, benzidine, 2- naphthylamines, naphthalidine are all the carcinogens of bladder transitional cell carcinoma, and occupational factor is led
The bladder transitional cell carcinoma patient of cause accounts for about the 25% of bladder transitional cell carcinoma patient populations.
Chromosome instability is fixed usually related to tumour, specifically includes the missing of whole chromosome or chromosome segment copy
Or amplification.Tumour is often containing the amplification and missing that relevant chromosome or chromosome segment occur to tumour, and institute spy occurs
Have, the detection to tumor staining body unstable region, research tumour is occurred and exploitation diagnosing tumor technology is all to pass
It is important.There is the method using Fluorescence in situ hybridization on Present clinical, unstability detection subregional for chromosome top, but
Lack the distribution characteristics of bladder transitional cell carcinoma patient's whole gene group level chromosome instability, more fully to instruct on urinary tract
The diagnosis and prognosis of skin cancer.
Summary of the invention
Present invention aims at research China's bladder transitional cell carcinoma (UC) specific to chromosome instability distribution characteristics,
The detailed process and chromosome instability for disclosing full-length genome two generations sequencing approach determine region, it was found that 10 and bladder transitional cell carcinoma
Occur common chromosomal variation region, total incidence accounts for 93.4% in bladder transitional cell carcinoma, for clinical early diagnosis in next step
Scientific basis is provided with individualized treatment scheme is formulated.
The first aspect of the present invention provides the chromosome instability often occurred in one group of China bladder transitional cell carcinoma patient and determines area
Domain, including following 10: 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q, 17p, 17q.
Wherein the overexpression of RAF1 is related to bladder cancer pathological analysis in the area 3p25；7p amplification leads to epidermal growth factor
The high expression of EGFR (proto-oncogene)；The missing of the site 9p16 tumor suppressor gene, leads to the exception of cell cycle regulating；17p missing is led
It causes tumor suppressor gene TP53 that cannot express, promotes tumour progression.
The second aspect of the present invention, provide a group chromosome unstable region preparation diagnosis bladder transitional cell carcinoma reagent or
Application in kit, the chromosome instability determine region include following 10: 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q,
17p、17q。
Further, the chromosome instability includes the missing or expansion of whole chromosome or chromosome segment copy surely
Increase.
Further, the bladder transitional cell carcinoma includes: carcinoma of renal pelvis, carcinoma of ureter, bladder cancer, urothelial cell cancer.
Third aspect present invention provides a group chromosome unstable region answering in preparation treatment urothelium cancer drug
With it includes following 10 that the chromosome instability, which determines region: 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q, 17p, 17q.
Fourth aspect present invention provides a group chromosome unstable region in the reagent for preparing bladder transitional cell carcinoma assessment prognosis
Or the application in kit, the chromosome instability determine region include following 10: 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q,
17p、17q。
Fifth aspect present invention provides a group chromosome unstable region in the reagent for preparing bladder transitional cell carcinoma early screening
Or the application in kit, the chromosome instability determine region include following 10: 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q,
17p、17q。
Sixth aspect present invention provides a group chromosome unstable region in the reagent for preparing bladder transitional cell carcinoma state of illness monitoring
Or the application in kit, the chromosome instability determine region include following 10: 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q,
17p、17q。
Seventh aspect present invention provides a kind of bladder transitional cell carcinoma screening, bladder transitional cell carcinoma diagnosis, bladder transitional cell carcinoma prognosis are commented
Estimate or the kit of bladder transitional cell carcinoma disease surveillance, include in kit: 1) detecting 10 chromosome instabilities as described above
Determine one or more reagents in region；2) one or more components selected from the group below: specification, positive reference product, negative reference
Product, buffer, enzyme, library connector, detectable label, nucleic acid extracting reagent, nucleic acid purification reagent.
Detection kit of the invention has the operation instructions of kit, includes how to carry out experiment behaviour in specification
Make, how according to data result to judge how whether above-mentioned 10 chromosomal regions occur unstable variation and result to urine
The diagnosis of road epithelioma, treatment, prognosis evaluation etc. are illustrated.
Using kit of the invention, above-mentioned 10 can be detected by various methods (including but not limited to) selected from the group below
Chromosomal region: digital pcr, Fluorescence in situ hybridization, nucleic acid probe hybridization method etc..
In addition, the kit also includes clinically for the screening of bladder transitional cell carcinoma, diagnosis, therapeutic scheme selection, prison
Other reagents with prognosis evaluation are surveyed, to assist or be verified the detection obtained result of above-mentioned 10 chromosomal regions.This
Field those of ordinary skill carries out conventional selection according to specific needs.
Using kit of the invention, selected using the screening, diagnosis, therapeutic scheme that following methods carry out bladder transitional cell carcinoma
It selects, monitor and prognosis evaluation: 1) obtaining sample to be tested from object；2) sample to be tested is contacted with using detection reagent of the invention；
3) above-mentioned 10 chromosomal regions of sample to be tested are detected；4) screening, diagnosis, the treatment of bladder transitional cell carcinoma are carried out according to testing result
Scheme Choice, monitoring and prognosis evaluation.
Eighth aspect present invention provides a kind of unique library connector with specific label for the sequencing of two generations, is used for
Exclude when machine is gone up in other types library and library of the present invention simultaneously the mistake because caused by library index sequence is same or similar.
Further, the nucleotide sequence of the library connector with specific label is respectively as follows:
Connector -1 (SEQ ID NO.1):
5'-CGTGATAGATCGGAAGAGCACACGTCTGAACTCCAGTC-3'；
Connector -2 (SEQ ID NO.2):
5’-ACACTCTTTCCCTACACGACGCTCTTCCGATCTATCACGT-3’。
Ninth aspect present invention provides a kind of library connector with specific label for the sequencing of two generations as described above
Application in the kit of preparation diagnosis bladder transitional cell carcinoma.
The technical solution adopted in the present invention:
To 198 bladder transitional cell carcinoma patients of definitive pathological diagnosis, the non-tumor patient of 103 urinary systems, 48 Healthy Peoples
Nucleic acid carries out the sequencing of two generations, analysis statistics tumor patient and control group in the difference of the fixed aspect of chromosome instability, passes through ROC song
Line delimit threshold value, to reach higher sensitivity and specificity.
The method that the present invention uses is sequenced for genome sequencing or target area targeted capture, passes through low depth high pass
Sequence is measured, chromosomal copy number is analyzed, identification occurs chromosome instability and determines region.This method is easy to operate, accuracy
Height, the chromosome instability that can be detected 0.1% are determined.Due to the high sensitivity of this method, it can be widely used for various clinical sample classes
Type, such as body fluid, tissue, cell.
Target area catching method includes but is not limited to probe hybrid method, PCR amplification method.
We are interrupted human genome using physics or enzymatic cleavage methods, or directly use the Cell-free DNA of fragmentation,
It repaired by end, add A, the connector of specific label is had in connection, by amplification plus for distinguishing sample index, obtained literary
Library.Then library is quantified, upper machine sequencing.
Upper machine data analysis is completed in four steps: (a) sequence alignment examines genome sequence to ginseng；(b) to coverage
Carry out G/C content and the correction of other weighting coefficients；(c) relative cover Z-score is calculated, the counting of test sample subtracts ginseng
According to sample counting, then the standard deviation covered divided by the sample for reference section；(d) Z-score is more than given threshold (such as 3), fixed
Justice is that target area expands, and Z-score is lower than given threshold (such as -3), is defined as target area loss.
It can be the reference sequences that NCBI or UCSC are provided that genome is referred in step a.Comparison software is any software, packet
Include freeware BWA, SOAPaligner, Bowtie etc., including business software OmicSoft etc..
In step b, weighting coefficient is the possible correction coefficient for calculating error of sequencing sequence feature, and copy number calculates
When (Z-score) be the factor for influencing copy number and calculating, including G/C content, if repetitive sequence etc., chromosome translocation (B-
To influence the factor that dystopy calculates when Score) calculating, simple repeated sequence and length are included whether, if genome is conservative
Sequence, if include STR sequence etc..
In step c, this preferred case, sample for reference (no less than 10) is healthy sample for reference.
By aforesaid operations, it was found that with the very high 10 common chromosome abnormality regions of bladder transitional cell carcinoma correlation, packet
Containing 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q, 17p, 17q.The overexpression of RAF1 is related to bladder cancer pathological analysis in the area 3p25；7p
Amplification leads to the high expression of epidermal growth factor EGFR (proto-oncogene)；The missing of the site 9p16 tumor suppressor gene, leads to the cell cycle
The exception of regulation；17p missing causes tumor suppressor gene TP53 that cannot express, and promotes tumour progression.
From the point of view of carrying frequency, 10 chromosome instabilities determine region and are up to 93.4% in bladder transitional cell carcinoma patient, amount to
185 patients carry above-mentioned chromosome instability and determine region.Wherein unstable most commonly seen with 8q, carrying rate is up to 55.8%；5p,
9q exception occurrence frequency is only second to 8q, is 46.5% and 38.6% respectively；Other chromosome abnormality incidences are respectively 3p
(37.6%), 3q (30.1%), 7p (35.4%), 7q (19.2%), 9p (38.6%), 17p (19.6%), 17q (16.9%).
By aforesaid operations, it was found that chromosome instability described in aforementioned present invention is determined region and do not examined in normal healthy controls
It is abnormal out.
Bladder cancer patients 154 in 198 bladder transitional cell carcinoma patients, ureter patient 13, carcinoma of renal pelvis patient 31, this
It is close to invent 10 chromosomal regions diagnosis performance in 3 kinds of uropoiesis epitheliomas, without obvious Preference.Pass through ROC curve
Compare 10 regions of the invention to show in whole uropoiesis epithelioma and bladder cancer, AUC is respectively 0.941 and 0.934, the two inspection
Similar performance is surveyed, referring in particular to embodiment 2 and attached drawing 2,3.
The invention has the advantages that:
The present invention collects the sample of the bladder transitional cell carcinoma patient of clinical definite, Healthy People and non-bladder transitional cell carcinoma Disease
This, by genome sequencing comparison of tumor patient and the chromosomal abnormalities compareed, illustrates chromosome specific to UC not
Stability distribution characteristics discloses 10 common chromosome instabilities of bladder transitional cell carcinoma patient and determines region, and total carrying rate is up to
93.4%, this has a very big significance the diagnosis of clinically bladder transitional cell carcinoma, treatment, monitoring meter prognosis evaluation, in next step
It is early diagnosed and is formulated individualized treatment scheme and scientific basis is provided.
Detailed description of the invention
Fig. 1 chromosome instability setting analysis result figure；Wherein: A, 3p+ and 3q+, 3q+, 5p+；B, 7p+, 7q+, 8q+；C,
9p-, 9q-, 9p- and 9q-；D, 17p-, 17q+.
Fig. 2 detection method ROC curve figure -- urothelial cancer.
Fig. 3 detection method ROC curve figure -- bladder cancer.
Specific embodiment
It elaborates below with reference to embodiment to specific embodiment provided by the invention.
Embodiment 1: the preparation of library connector
Via third company's synthetic linker sequence:
Connector -1 (SEQ ID NO.1):
5'-CGTGATAGATCGGAAGAGCACACGTCTGAACTCCAGTC-3'；
Connector -2 (SEQ ID NO.2):
5’-ACACTCTTTCCCTACACGACGCTCTTCCGATCTATCACGT-3’。
5 ' ends of connector -1 carry out phosphorylation modification, carry out thiophosphoric acid modification between 3 ' end G and T of connector -2.
The dry powder of connector -1 and connector -2 is diluted to 20 μM respectively using annealing buffer (Annealing buffer),
The two dilution equal proportion is mixed, by mixed liquor as in PCR instrument, runs program such as the following table 1:
Table 1
Product is subjected to agarose electrophoresis, electrophoretic band magnitude range is between 80-100bp.
Embodiment 2:
Use material:
Human gene group DNA
DNA enzymatic, which is cut, interrupts reagent (Enzymatics)
The end DNA is repaired plus A, adjunction head enzyme reaction system (NEB)
Pcr amplification reaction system (Kapa Biosystems)
It purifies magnetic bead (Beckman)
Operating procedure:
1, genomic fragment
20ng genomic DNA is taken, using endonuclease reaction system, reaction system is configured according to table 2, is reacted according to table 3.
Is needed not move through by genomic fragment, is directly entered down such as the cfDNA in blood for cell-free dissociative DNA
Step operation.
Table 2
Oscillation mixes, runs following procedure in PCR instrument after centrifugation (avoiding bubble):
Table 3
2, end, which is repaired, adds A
Add A reaction system such as the following table 4 with set terminal reparation:
Table 4
Oscillation mixes, runs following procedure in PCR instrument after centrifugation (avoiding bubble):
Table 5
3, adjunction head and product purification
Connector is connected into premixed liquid (30 μ L/ sample), connector link enhancement agent (1 μ L/ sample) according to sample detected
After number absorption volume appropriate is mixed, the end-o f-pipe -control reaction mixture that 15.5 μ L mixed liquors are added to previous step is drawn
In, the connector prepared in 2.5 μ L/ embodiments 1 is drawn again after oscillation mixing, centrifugation is added in above-mentioned reaction solution, oscillation mixing,
Centrifugation.Ultimately form the reaction system in the following table 6.
Table 6
| Connector connection reaction | System |
| Repair reaction mixture (previous step) in end | 60μL |
| Connector connects premixed liquid | 30μL |
| Connector link enhancement agent | 1μL |
| Connector | 2.5μL |
| Total system | 93.5μL |
Above-mentioned system is placed in PCR instrument, run program: 20 DEG C, 30min, heat lid is closed.
Purification step after connection reaction:
A, library is taken out from magnetic bead kit in advance and purify magnetic bead, be stored at room temperature at least 30min, need to mix before use.
B, the connector connection reaction solution in above-mentioned steps is transferred to the 1.5mL centrifuge tube accordingly numbered, 86 μ L weight is added
Outstanding library purifies magnetic bead, is at the uniform velocity inhaled and is made a call to 20 times using the pipettor of appropriate range, is incubated at room temperature 5min.
C, centrifuge tube is placed on magnetic frame, after solution clarification, abandons supernatant liquor.
D, the ethyl alcohol of 200 μ L 80% of fresh configuration is added thereto, after standing 30s, abandons supernatant liquor.
E, it is primary to repeat step d.
F, centrifuge tube, brief centrifugation 3sec are removed on magnetic frame, centrifuge tube is put back on magnetic frame, and suction discards remaining 80%
Ethyl alcohol is careful not to be drawn onto magnetic bead.Pipe lid is opened, room temperature dries 2-10min.
G, to magnetic bead at matt color, 16 μ L Low TE buffers (or nuclease-free water) are added in centrifuge tube, it is slight to shake
Swinging is resuspended magnetic bead, is incubated at room temperature 5min.
H, centrifuge tube is placed on magnetic frame, stands 2min.After solution clarification, 15 μ L supernatants is taken to remain to expand in next step
Reaction.
4, PCR amplification and product purification
According to the following table 7, corresponding reagent is added into PCR pipe:
Table 7
| PCR reaction | System |
| Connector connects product after purification | 15μL |
| PCR amplification enzyme premixed liquid (green | 25μL |
| P covers C) R amplification universal primer (green | 5μL |
| P covers C) R amplification Tag primer | 5μL |
| Total system | 50μL |
The PCR pipe mixed is put into PCR instrument, following procedure is run.
Table 8
PCR product purifying:
A, library is taken out from magnetic bead kit in advance and purify magnetic bead, be stored at room temperature at least 30min, need to mix before use.
B, the connector connection reaction solution in above-mentioned steps is transferred to the 1.5mL centrifuge tube accordingly numbered, 22.5 μ L is added
The library of resuspension purifies magnetic bead, is at the uniform velocity inhaled and is made a call to 20 times using the pipettor of appropriate range, is incubated at room temperature 5min.
C, centrifuge tube is placed on magnetic frame, after solution clarification, abandons supernatant liquor.
D, the ethyl alcohol of 200 μ L 80% of fresh configuration is added thereto, after standing 30s, abandons supernatant liquor.
E, it is primary to repeat step d.
F, centrifuge tube, brief centrifugation 3sec are removed on magnetic frame, centrifuge tube is put back on magnetic frame, and suction discards remaining 80%
Ethyl alcohol is careful not to be drawn onto magnetic bead.Pipe lid is opened, room temperature dries 2-10min.
G, to magnetic bead at matt color, 31 μ L Low TE buffers (or nuclease-free water) are added in centrifuge tube, it is slight to shake
Swinging is resuspended magnetic bead, is incubated at room temperature 5min.
H, centrifuge tube is placed on magnetic frame, stands 2min.After solution clarification, 30 μ L supernatants is taken to remain to expand in next step
Reaction.
5, machine is quantitatively gone up in library
Above-mentioned purified library analyzes clip size using Agilent BioAnalyzer, uses
DsDNA HS Assay Kit measures Library Quality concentration, and it is dense to calculate library mole by mass concentration and clip size
Degree, is sequenced according to upper sequenator specification using Illumina Hiseq X-ten.
Interpretation of result discussion:
Upper machine sequencing data obtains chromosome sequencing depth profile, such as after data analysing method of the present invention
Shown in Fig. 1.Analysis result consists of three parts: depth distribution, dyeing is sequenced in chromosome numbers, chromosome subregion and chromosome
Body be sequenced depth distribution region orbicular spot be chromosome zonule copy number distribution situation, when its in longitudinal axis scores value-3-
Determine to be judged to expanding when its scores value is greater than 3, sentence when its scores value is less than -3 without unstable generation when between 3
It is set to missing, the region for occurring to expand or lack is distinguished using gray background and normal region.By analyzing result
It has checked whether that above-mentioned chromosome instability occurs surely, if being determined as the positive, if not being determined as feminine gender, two generations was analyzed
As a result it is compared with pathological examination, calculates the sensitivity and specificity of this analysis method, obtain ROC curve, such as Fig. 2,3 institutes
Show.The AUC area of this method ROC curve reaches 0.941, illustrates using 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q, 17p, 17q not
Stablize as analysis mark, diagnosis urothelial cancer, which is sequenced, by two generations has very high sensitivity and specificity.In wing
AUC is 0.934 in Guang cancer, and the two detection performance is close.
The preferred embodiment of the present invention has been described in detail above, but the invention be not limited to it is described
Embodiment, those skilled in the art can also make various equivalent on the premise of not violating the inventive spirit of the present invention
Variation or replacement, these equivalent variation or replacement are all included in the scope defined by the claims of the present application.
Sequence table
<110>Suzhou Hong Yuan Biotechnology Co., Ltd
The unstable variation of<120>one group chromosomes is in preparation diagnosis bladder transitional cell carcinoma, the reagent or kit of assessing prognosis
Using
<130> /
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 38
<212> DNA
<213>artificial sequence (Artificial)
<400> 1
cgtgatagat cggaagagca cacgtctgaa ctccagtc 38
<210> 2
<211> 40
<212> DNA
<213>artificial sequence (Artificial)
<400> 2
acactctttc cctacacgac gctcttccga tctatcacgt 40
Claims (10)
1. application of the group chromosome unstable region in the reagent or kit of preparation diagnosis bladder transitional cell carcinoma, the dye
Colour solid unstable region includes following 10: 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q, 17p, 17q.
2. group chromosome unstable region according to claim 1 is in the reagent or reagent of preparation diagnosis bladder transitional cell carcinoma
Application in box, which is characterized in that the chromosome instability include surely whole chromosome or chromosome segment copy lack
It loses or expands.
3. group chromosome unstable region according to claim 1 is in the reagent or reagent of preparation diagnosis bladder transitional cell carcinoma
Application in box, which is characterized in that the bladder transitional cell carcinoma includes: that carcinoma of renal pelvis, carcinoma of ureter, bladder cancer, urothelial are thin
Born of the same parents' cancer.
4. application of the group chromosome unstable region in preparation treatment urothelium cancer drug, the chromosome instability are fixed
Region includes following 10: 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q, 17p, 17q.
5. application of the group chromosome unstable region in the reagent or kit for preparing bladder transitional cell carcinoma assessment prognosis, described
Chromosome instability determine region include following 10: 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q, 17p, 17q.
6. application of the group chromosome unstable region in the reagent or kit for preparing bladder transitional cell carcinoma early screening, described
Chromosome instability determine region include following 10: 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q, 17p, 17q.
7. application of the group chromosome unstable region in the reagent or kit for preparing bladder transitional cell carcinoma state of illness monitoring, described
Chromosome instability determine region include following 10: 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q, 17p, 17q.
8. a kind of bladder transitional cell carcinoma screening, bladder transitional cell carcinoma diagnosis, bladder transitional cell carcinoma prognosis evaluation or bladder transitional cell carcinoma disease
The kit of monitoring includes: 1) detecting one or more reagents that 10 chromosome instabilities determine region；2) one kind selected from the group below
Or various ingredients: specification, positive reference product, negative reference product, buffer, enzyme, library connector, detectable label, nucleic acid mention
Take reagent, nucleic acid purification reagent；10 chromosome instabilities determine region and include: 3p, 3q, 5p, 7p, 7q, 8q, 9p, 9q,
17p、17q。
9. a kind of library connector with specific label for the sequencing of two generations, which is characterized in that described has specific label
Library connector nucleotide sequence as shown in SEQ ID NO.1 or SEQ ID NO.2.
10. a kind of library connector with specific label as claimed in claim 9 is in the reagent of preparation diagnosis bladder transitional cell carcinoma
Application in box.
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| CN111254198A (en) * | 2020-03-09 | 2020-06-09 | 苏州宏元生物科技有限公司 | Kit for diagnosing and prognostically evaluating urothelial cancer |
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| Publication number | Priority date | Publication date | Assignee | Title |
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