CN109355393A - Cervical cell smearing relevant HPV integrator gene site and its application - Google Patents
Cervical cell smearing relevant HPV integrator gene site and its application Download PDFInfo
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- CN109355393A CN109355393A CN201811519133.4A CN201811519133A CN109355393A CN 109355393 A CN109355393 A CN 109355393A CN 201811519133 A CN201811519133 A CN 201811519133A CN 109355393 A CN109355393 A CN 109355393A
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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
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57411—Specifically defined cancers of cervix
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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
- C12Q2600/158—Expression markers
Abstract
The invention discloses a kind of for diagnosing the biomarker of cervical cell smearing, the biomarker is following gene family: MYC gene family, NR4A gene family or ANKRD gene family or combinations thereof, or its encoded albumen, wherein, the biomarker is integrated with the partial sequence or full sequence of HPV.The invention also discloses containing the biomarker kit and its application.Biomarker of the invention or containing its kit have the advantages that in the application of detection cellule cervical carcinoma quickly, precisely, conveniently, false positive rate it is low etc., with important clinical diagnosis meaning.
Description
Technical field
The present invention relates to biological diagnosis field more particularly to a kind of cervical cell smearing relevant HPV integrator gene sites
And its application.
Background technique
Cervical carcinoma is the second largest reason of whole world young and middle-aged women (20-39 years old) cancer mortality, histopathology type
For squamous carcinoma, gland cancer, adenosquamous carcinoma and rare refractory type cervical carcinoma (such as: cervical cell smearing, uterine neck clear cell carcinoma).With
Further investigation and nearly more than 10 year of the gradual perfection, scientist of the medical support system of early screening to cervical cancer pathogenesis mechanism
The popularization and application of human papilloma virus (HPV) vaccine, global cervical cancer pathogenesis rate just decline year by year.Duration high-risk-type human milk head
Tumor virus (HR-HPV) infects the major virulent factor for being acknowledged as uterine neck carcinogenesis.However in recent years, China's cervical cancer pathogenesis
Rate but obviously rebounds and the onset peak age tends to rejuvenation, possible cause are as follows: 1. gradually opening with our people's attitude towards sex
It puts, HPV infection is more spread unchecked；2.HPV vaccine is in the granted Time To Market in China mainland area later (divalent, tetravalence HPV vaccine point
Just do not get the Green Light in July, 2016, in May, 2017 in clinical use) and HPV vaccine be not yet included in ordinarily resident's medical insurance
Scope.The HPV vaccine inoculation time that FDA recommends is 9-26 years old.Obviously, large quantities of Chinese women have been over injection cervical carcinoma epidemic disease
The recommended age of seedling, they still have high risk to suffer from cervical carcinoma in the coming few decades that do not protected by vaccine.Therefore, exist
China, cervical carcinoma are still the major disease for needing actively to study.Underwent operative+chemicotherapy Canonical management, the common class of early stage
5 years survival rates of type cervical carcinoma (squamous carcinoma, gland cancer) can reach about 70%.However, some rare refractory type cervical carcinomas, by
In its disease incidence is extremely low, pathogenesis is unknown, it is high, insensitive to chemicotherapy to be easy Misdiagnosis and grade of malignancy, survival rate according to
It is not effectively improved so, poor prognosis, it would be highly desirable to tackle key problems.
Cervical cell smearing (Small-Cell Carcinoma of the Cervix, SCCC) is a kind of rare but pernicious
Strong special cervical carcinoma type, belongs to one kind of Neuroendocrine Cervical Carcinoma, and disease incidence is aggressive cervical carcinoma
1%-2%.It is compared with cervical squamous cell carcinoma with gland cancer, SCCC can occur between the higher lymph node rate of transform and blood vessel in disease early stage
Gap infiltration rate, tumor recurrence will occur rapidly to Most patients in the several months after the treatment.The diagnostic criteria and therapeutic scheme of SCCC
With reference to the diagnosing and treating experience of small cell carcinoma of lung: diagnostic criteria is morphology (tumour is in small circular under mirror)+immune group
Change dyeing (at least one neuroendocrine index is positive, such as CgA, NSE, Syn, CD56)；Standard regimens are+putting of performing the operation
The complex treatment for the treatment of.But above scheme is poor to the curative effect of SCCC, survival rate is less than 14% within 5 years.Due to its rare property, greatly
Report of the part in relation to SCCC is case management report and small-scale clinical observation on the therapeutic effect, there is no causing for SCCC for authority at present
The basic research of interpretation of the cause, onset and process of an illness system, HPV integrated state and integrated mechanism.
Previously development HPV integration site detection method have Southern hybridization (CullenAP etc., JVirol, 1991；
65:606-612), integrate HPV viruse oncogene transcripton amplification (KlaesR etc., CancerRes, 1999；59:6132-
6136), restriction enzyme site PCR (ThorlandEC etc., CancerRes, 2000；60:5916-5921), real-time quantitative PCR
(PeitsaroP etc., JClinMicrobiol, 2002；40:886-891) and DNA in situ hybridization (EvansMF etc., JPathol,
2004；202:1-4).Although these methods can detect HPV integration site, exist susceptibility it is low, operation it is numerous
Trivial, time-consuming, a large amount of sample of need judges the disadvantages of uncertain to result.With the rapid development of high throughput sequencing technologies, capture
Sequencing and genome sequencing become precisely detection HPV integration effective ways (HuZ etc., NatGenet, 2015,47:158-
63)。
Currently, detection method and product for High-risk Population of Cervical Carcinoma primarily directed to whether there is or not high-risk HPV infect into
Row detection, but and not all high-risk HPV the infected cervical carcinoma all has occurred, detected at present for the integration site of HPV
The method and product of common type High-risk Population of Cervical Carcinoma are researching and developing (CN104830869A), there is no detection uterine neck cellule
The research of cancer HPV integration site and product.
Summary of the invention
The information that the present invention overcomes this field conventional detection cervical cell smearing that can not know whether that HPV is integrated lacks
It falls into, provides a kind of biomarker for diagnosing cervical cell smearing or kit and its application containing it.Of the invention
Biomarker combinations and containing its kit have the advantages that quickly, precisely, conveniently, false positive rate it is low etc., there is important face
Bed diagnostic significance.
In order to solve the above technical problems, one of technical solution of the present invention is, for diagnosing the biology of cervical cell smearing
Marker, the biomarker be following gene family: MYC gene family, NR4A gene family or ANKRD gene family or
A combination thereof or its encoded albumen, wherein the biomarker is integrated with the partial sequence or full sequence of HPV.This hair
In bright, " partial sequence " refers to the sequence fragment of the HPV of at least one nucleotide shorter than " complete sequence " or amino acid.
In one embodiment, the MYC gene family includes MYC gene and MYCN gene；NR4A gene man
Race includes NR4A2 gene；The ANKRD gene family includes ANKD12 gene.
In a preferred embodiment, the HPV is HPV16 and/or HPV18.
In one embodiment, the biomarker integrates the partial sequence or full sequence of HPV.It is preferably real one
It applies in example, the partial sequence or full sequence for integrating HPV are as shown in SEQ ID NO:97 to SEQ ID NO:144.
In one embodiment, the biomarker integrates the partial sequence of HPV or full sequence generates breakpoint, institute
Stating breakpoint includes the breakpoint such as below table on HPV breakpoint and human genome
It is shown.
In SCCC tumour, other than the biomarker of HPV of the present invention integration, there are also the reports of other biological marker
Road, by it has been reported that biomarker together with biomarker combinations of the invention, SCCC will be better diagnosed.Cause
This, in a preferred embodiment, the biomarker further includes INSM1 and/or ASCL1.
In one embodiment, above-described biomarker can be detected by WGS, VCS or immunohistochemistry.
In order to solve the above technical problems, the two of technical solution of the present invention are, prepared by biomarker as described above
Purposes in the diagnosis marker of cervical cell smearing or the kit containing it.
In order to solve the above technical problems, the three of technical solution of the present invention are, a kind of reagent diagnosing cervical cell smearing
Box, the kit include the reagent for detecting the biomarker as described in claim any one of 1-7.Preferably, described
Reagent is primer.It is highly preferred that the amplified production of the primer is the sequence as shown in SEQ ID NO:97 to SEQ ID NO:144
Column.
Most preferably, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:1, and downstream nucleotide sequence is such as
Shown in SEQ ID NO:49；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:2, downstream nucleotide sequence
As shown in SEQ ID NO:50；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:3, downstream nucleotide sequence
Column are as shown in SEQ ID NO:51；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:4, downstream nucleotide
Sequence is as shown in SEQ ID NO:52；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:5, downstream nucleotide
Acid sequence is as shown in SEQ ID NO:53；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:6, downstream core
Nucleotide sequence is as shown in SEQ ID NO:54；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:7, downstream
Nucleotide sequence is as shown in SEQ ID NO:55；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:8, under
Nucleotide sequence is swum as shown in SEQ ID NO:56；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:9,
Downstream nucleotide sequence is as shown in SEQ ID NO:57；Or, the upstream nucleotide sequence of the primer such as SEQ ID NO:10 institute
Show, downstream nucleotide sequence is as shown in SEQ ID NO:58；Or, the upstream nucleotide sequence of the primer such as SEQ ID NO:11
Shown, downstream nucleotide sequence is as shown in SEQ ID NO:59；Or, the upstream nucleotide sequence of the primer such as SEQ ID NO:
Shown in 12, downstream nucleotide sequence is as shown in SEQ ID NO:60；Or, the upstream nucleotide sequence of the primer such as SEQ ID
Shown in NO:13, downstream nucleotide sequence is as shown in SEQ ID NO:61；Or, the upstream nucleotide sequence of the primer such as SEQ
Shown in ID NO:14, downstream nucleotide sequence is as shown in SEQ ID NO:62；Or, the upstream nucleotide sequence of the primer is such as
Shown in SEQ ID NO:15, downstream nucleotide sequence is as shown in SEQ ID NO:63；Or, the upstream nucleotide sequence of the primer
As shown in SEQ ID NO:16, downstream nucleotide sequence is as shown in SEQ ID NO:64；Or, the nucleotides upstream sequence of the primer
Column are as shown in SEQ ID NO:17, and downstream nucleotide sequence is as shown in SEQ ID NO:65；Or, the nucleotides upstream of the primer
Sequence is as shown in SEQ ID NO:18, and downstream nucleotide sequence is as shown in SEQ ID NO:66；Or, the upstream nucleosides of the primer
Acid sequence is as shown in SEQ ID NO:19, and downstream nucleotide sequence is as shown in SEQ ID NO:67；Or, the upstream core of the primer
Nucleotide sequence is as shown in SEQ ID NO:20, and downstream nucleotide sequence is as shown in SEQ ID NO:68；Or, the upstream of the primer
Nucleotide sequence is as shown in SEQ ID NO:21, and downstream nucleotide sequence is as shown in SEQ ID NO:69；Or, the primer is upper
Nucleotide sequence is swum as shown in SEQ ID NO:22, downstream nucleotide sequence is as shown in SEQ ID NO:70；Or, the primer
Upstream nucleotide sequence is as shown in SEQ ID NO:23, and downstream nucleotide sequence is as shown in SEQ ID NO:71；Or, the primer
Upstream nucleotide sequence as shown in SEQ ID NO:24, downstream nucleotide sequence is as shown in SEQ ID NO:72；Or, described draw
The upstream nucleotide sequence of object is as shown in SEQ ID NO:25, and downstream nucleotide sequence is as shown in SEQ ID NO:73；Or, described
The upstream nucleotide sequence of primer is as shown in SEQ ID NO:26, and downstream nucleotide sequence is as shown in SEQ ID NO:74；Or, institute
The upstream nucleotide sequence of primer is stated as shown in SEQ ID NO:27, downstream nucleotide sequence is as shown in SEQ ID NO:75；Or,
The upstream nucleotide sequence of the primer is as shown in SEQ ID NO:28, and downstream nucleotide sequence is as shown in SEQ ID NO:76；
Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:29, downstream nucleotide sequence such as SEQ ID NO:77 institute
Show；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:30, downstream nucleotide sequence such as SEQ ID NO:78
It is shown；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:31, downstream nucleotide sequence such as SEQ ID NO:
Shown in 79；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:32, downstream nucleotide sequence such as SEQ ID
Shown in NO:80；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:33, downstream nucleotide sequence such as SEQ
Shown in ID NO:81；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:34, downstream nucleotide sequence is such as
Shown in SEQ ID NO:82；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:35, downstream nucleotide sequence
As shown in SEQ ID NO:83；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:36, downstream nucleotide sequence
Column are as shown in SEQ ID NO:84；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:37, downstream nucleotide
Sequence is as shown in SEQ ID NO:85；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:38, downstream nucleotide
Acid sequence is as shown in SEQ ID NO:86；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:39, downstream core
Nucleotide sequence is as shown in SEQ ID NO:87；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:40, downstream
Nucleotide sequence is as shown in SEQ ID NO:88；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:41, under
Nucleotide sequence is swum as shown in SEQ ID NO:89；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:42,
Downstream nucleotide sequence is as shown in SEQ ID NO:90；Or, the upstream nucleotide sequence of the primer such as SEQ ID NO:43 institute
Show, downstream nucleotide sequence is as shown in SEQ ID NO:91；Or, the upstream nucleotide sequence of the primer such as SEQ ID NO:44
Shown, downstream nucleotide sequence is as shown in SEQ ID NO:92；Or, the upstream nucleotide sequence of the primer such as SEQ ID NO:
Shown in 45, downstream nucleotide sequence is as shown in SEQ ID NO:93；Or, the upstream nucleotide sequence of the primer such as SEQ ID
Shown in NO:46, downstream nucleotide sequence is as shown in SEQ ID NO:94；Or, the upstream nucleotide sequence of the primer such as SEQ
Shown in ID NO:47, downstream nucleotide sequence is as shown in SEQ ID NO:95；Or, the upstream nucleotide sequence of the primer is such as
Shown in SEQ ID NO:48, downstream nucleotide sequence is as shown in SEQ ID NO:96.
In a preferred embodiment, the kit further includes that DNA extracts reagent and PCR amplification reagent.
On the basis of common knowledge of the art, above-mentioned each optimum condition, can any combination to get each preferable reality of the present invention
Example.
The reagents and materials used in the present invention are commercially available.
The positive effect of the present invention is that:
1, using biomarker of the invention or containing its kit, can auxiliary diagnosis cervical cell smearing, or
The precancerous lesion crowd having progressed as cervical cell smearing risk is sifted out to carry out emphasis monitoring.
2, when carrying out the general sieve of cervical carcinoma in crowd, can more have using biomarker of the invention or containing its kit
Cervical cell smearing patient and people at highest risk are targetedly sifted out, workload more delicately can be diagnosed or reduce.
To sum up, of the present invention to utilize HPV integration site for biological marker detection cellule High-risk Population of Cervical Carcinoma
Marker combination and containing its kit have the advantages that quickly, precisely, conveniently, false positive rate it is low etc., with important clinic
Diagnostic significance.
Detailed description of the invention
The ratio of integrator gene family of HPV viruse institute in 150 samples is reflected in Fig. 1 SCCC in HPV confluence analysis figure.
Specific embodiment
The present invention is further illustrated below by the mode of embodiment, but does not therefore limit the present invention to the reality
It applies among a range.In the following examples, the experimental methods for specific conditions are not specified, according to conventional methods and conditions, or according to quotient
The selection of product specification.
1 sample of embodiment prepares and processing
Tissue DNA extracts kit (QIAamp DNA Mini Kit) is bought from QIAGEN company.
Select 16 cervical cell smearing patient cancer stove tissues and cancer side control tissue as sample.
1) 25 milligrams of cervical tissue choppings are taken, is placed in 1.5 milliliters of pipes, 180 microlitres of solution B uffer ATL is added, shake
It mixes；
2) 20 microlitres of Proteinase Ks (Proteinase K), concussion are added, 56 DEG C of water-baths are completely dissolved until cervical tissue；
3) low-speed centrifugal gets rid of lower tube top liquid, adds 5 microlitre of 100 mg/ml RNAase A, shakes 15 seconds；
4) be added 200 microlitres of solution B uffer AL, shake 15 seconds, 70 DEG C water-bath 10 minutes；
5) 200 microlitres of dehydrated alcohols, interruption concussion 15 seconds is added in brief centrifugation；Brief centrifugation is transferred to 2 milliliters of Spin
Column, revolving speed are 8000 revs/min, are centrifuged 1 minute, abandon centrifugal liquid；
6) 500 microlitres of solution B uffer AW1 are added, revolving speed is 8000 revs/min, is centrifuged 1 minute, abandons centrifugal liquid；
7) solution B uffer AW2 is added, revolving speed is 14000 revs/min, is centrifuged 3 minutes, abandons centrifugal liquid；
8) Spin column adsorption column is dallied 1 minute, alcohol of sufficiently leaving away；
9) Buffer AE eluted dna is added, the DNA of elution is stored in 1.5 milliliters of pipes, is stored in -80 DEG C of refrigerators, obtains
Obtain the genomic DNA of the cervical tissue of 1 microgram.
The integration detection of 2 high throughput virus of embodiment
WGS sequencing is carried out to 16 pairs of Fresh Frozen tissues.
The genome DNA sample of the cervical tissue extracted in embodiment 1 is taken out from -80 DEG C of refrigerators, is placed on added with dry
In the incubator of ice, it is sent to Shenzhen Hua Da Gene science Co., Ltd and carries out WGS sequencing.The following are 16 couples of sample WGS to survey
Sequence is as a result, the results show that HPV16/18 is incorporated into gene M YC, MYCN, NR4A2 and ANKD12, sample percentage such as table 1
It is shown.After table 2 shows HPV genome insertion human genome, the breakpoint of human genome and HPV.
Table 1.16 is to sample WGS sequencing result
| Gene | Sample percentage |
| Gene M YC | 31.25% |
| Gene M YCN | 6.25% |
| Gene family MYC is total | 37.5% |
| Gene NR4A2 | 6.25% |
| Gene A NKD12 | 6.25% |
Embodiment 3 utilizes PCR amplification and Sanger sequence verification HPV integration site
Design expands the upstream and downstream primer of the integration site, prepares reaction system by following table 2 system.
Table 2.PCR reaction system
Every pipe mixes respectively, in the PCR amplification instrument of standard, is expanded using 3 program of table: table 3.PCR amplification program
The PCR product obtained after amplification is sent to company and carries out Sanger sequencing.
The integration information and primer sequence of related gene are as shown in the table.
4. biomarker integration site of table, integration sequence and primer sequence
4 high-flux sequence method of embodiment verifies a large amount of clinical samples
HPV viruse capture sequencing (VCS) is successfully carried out to 150 FFPE samples, is carried out HPV confluence analysis (see Fig. 1)
Specifically, HPV integrates hot spot significant enrichment in 3 gene families, and closely related with tumour generation, point
Not are as follows: (sample percentage 37.04% is only that 9.7%), (sample percentage is for NR4A family in squamous carcinoma gland cancer for MYC family
6.17%), ANKRD family (sample percentage 4.94%).It is interesting that HPV 18 tends to be incorporated into MYC family, and
HPV 16 tends to be incorporated into NR4A family gene (P < 0.01, Fig. 1).
Integration of the table 5.HPV in larger scale clinical samples
| Gene family | Sample percentage |
| Gene family MYC | 37.04% |
| Gene family NR4A | 6.17% |
| Gene family ANKRD | 4.94% |
After the insertion of HPV genome, the breakpoint and primer sequence of human genome and HPV are referring to table 4.
Present invention discover that: HPV18 is most important infection type (83.3%) in SCCC, while being also most important integration
Type (63.3%).There were significant differences for the above results and the most common cervical squamous cell carcinoma, and (HPV16 is main infection and whole in squamous carcinoma
Mould assembly is other).According to the literature, HPV18 (such as cervical intraepithelial neoplasia (CIN) and cervical carcinoma group in different cervical lesions tissues
Knit) recall rate there was no significant difference, therefore the relationship of HPV18 and SCCC be present invention firstly discovers that, and its relevance and its
Its index is good many compared to effect.
The positive rate of 5 clinical sample of embodiment
Goldstandard-pathological diagnosis: 1. forms exactly like Small Cell Lung Cancer, and cell volume is small, in the more consistent short shuttle of size
Shape or round cell, endochylema be few, nuclear hyperchromatism, chromatin are in coarse granule shape, no kernel or kernel are unobvious, and mitosis figures are common.
2. immunohistochemistry at least expresses a kind of neuroendocrine markers, such as: CgA, Syn, CD56 and NSE (Albores-Saavedra
J,Gersell D,Gilks CB,et al.Terminology of endocrine tumors of the uterine
cervix:results of a workshop sponsored by the College of American
Pathologists and the National Cancer Institute.Arch Pathol Lab Med.1997Jan；
121(1):34-9.Kajiwara H,Hirabayashi K,Miyazawa M,et al.Immunohistochemical
expression of somatos tat in type 2A receptor in neuroendocrine carcinoma of
Uterine cervix.Arch Gynecol Obstet, 2009,279 (4): 521~525).
With in the SCCC sample of goldstandard detection, integration site true positives recall rate as described in example 4 is 50% (WGS
Data) and 47% (VCS data), and in general type cervical carcinoma, positive rate is only 10%.It can be seen that life of the invention
Object marker can be combined with goldstandard detection method, since goldstandard method and step is cumbersome, time cost is high, first use this
The biomarker of invention significantly reduces the operating time.In addition it is also possible to be used clarifying a diagnosis as in the sample of cervical carcinoma
This kit assists diagnosis its hypotype SCCC.Existing diagnostic means are that specific immunohistochemistry index is positive, our invention
To assist diagnosis the type cervical carcinoma to provide new method.
In addition, using biological marker of the invention in the sample for having been detected by cervical carcinoma or precancerous lesion (CIN)
Object or kit containing it, can auxiliary diagnosis cervical cell smearing, or sift out and have progressed as cervical cell smearing risk
Precancerous lesion crowd is to carry out emphasis monitoring.Because the precancerous lesion patient suffers from cancer risk with higher, it should early stage
Intervened, such as row conization, and existing guide to the classification diagnosis and treatment of precancerous lesion mainly from Pathomorphology angle
Degree sets out, not from molecular biology change angle illustrate which precancerous lesion people can continue observe follow-up, which need
Want immediate intervention.In addition, when carrying out the general sieve of cervical carcinoma in crowd more targeted palace can be sifted out using the kit
Neck small cell carcinoma patient and people at highest risk, more delicately can diagnose or reduce workload, and the method for traditional detection SCCC
It can not really know the information of HPV integration.
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, it is all in spirit of the invention and
Within principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
SEQUENCE LISTING
<110>Wuhan Kai Deweisi Bioisystech Co., Ltd
<120>cervical cell smearing relevant HPV integrator gene site and its application
<130> P180113917C
<160> 144
<170> PatentIn version 3.5
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<400> 17
acccttcttt cctccactct 20
<210> 18
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 18
<400> 18
aacccaacaa tagcagaagg 20
<210> 19
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 19
<400> 19
tggtgggata catgacaatg 20
<210> 20
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 20
<400> 20
tctaactgct gcttgtgtgg 20
<210> 21
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 21
<400> 21
ttccggttga ccttctatgt 20
<210> 22
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 22
<400> 22
ccgtccatta ttgaagttcc 20
<210> 23
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 23
<400> 23
atccccttga acattcacac 20
<210> 24
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 24
<400> 24
ggagacacac ctgagtggat 20
<210> 25
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 25
<400> 25
caaaaataga tgaaggggga 20
<210> 26
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 26
<400> 26
cgtctacata ggcctgcttt 20
<210> 27
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 27
<400> 27
gaaacatcac ccttgtcctc 20
<210> 28
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 28
<400> 28
gatcacagta gtggttgcca 20
<210> 29
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 29
<400> 29
attagcagac agcaacagca 20
<210> 30
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 30
<400> 30
aacatacacc tccagcacct 20
<210> 31
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 31
<400> 31
gaggaagagg aagatgcaga 20
<210> 32
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 32
<400> 32
ccatatttta gggttcctgc 20
<210> 33
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 33
<400> 33
caactaaaat gccctccatt 20
<210> 34
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 34
<400> 34
ttccacaagg cttttctagg 20
<210> 35
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 35
<400> 35
gcagagctag agacagcaca 20
<210> 36
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 36
<400> 36
attgcgaagt agtgttgcag 20
<210> 37
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 37
<400> 37
aaccacctat gtttactcag ga 22
<210> 38
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 38
<400> 38
aaaatcaacc cctcctcttc 20
<210> 39
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 39
<400> 39
ctgaagtgga agcaacacag 20
<210> 40
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 40
<400> 40
accacgtgtt ggacatactt t 21
<210> 41
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 41
<400> 41
cacctacagg caacaacaaa 20
<210> 42
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 42
<400> 42
acccacgcaa catttagttt 20
<210> 43
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 43
<400> 43
ttctcgctct gatacattcg 20
<210> 44
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 44
<400> 44
ttactgcaca ggaagcaaaa 20
<210> 45
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 45
<400> 45
ccctggaaat aactgagcac 20
<210> 46
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 46
<400> 46
gatagtgaat ggcaacgtga 20
<210> 47
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 47
<400> 47
acaagcaatt gaactgcaac 20
<210> 48
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>upstream primer 48
<400> 48
aaacgatgga gactctttgc 20
<210> 49
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 1
<400> 49
attgttatga ccctgtgcct 20
<210> 50
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 2
<400> 50
attgtgtgac gttgtggttc 20
<210> 51
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 3
<400> 51
ccaatctgta cacgtggttt 20
<210> 52
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 4
<400> 52
ccaaaacccc atgataagac 20
<210> 53
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 5
<400> 53
tccttgatga acattgatgc 20
<210> 54
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 6
<400> 54
gaaagattac cctcccaggt 20
<210> 55
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 7
<400> 55
cctcaggctg attcaaactt 20
<210> 56
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 8
<400> 56
gagtgattat caaggtgggc 20
<210> 57
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 9
<400> 57
aggggttgca atcctagtct 20
<210> 58
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 10
<400> 58
gagaaaaatg cagggacaat 20
<210> 59
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 11
<400> 59
agttccaggt ctgtgctagg 20
<210> 60
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 12
<400> 60
gatacgttta gaaggcattg g 21
<210> 61
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 13
<400> 61
gacaactgac tcccaaatcc 20
<210> 62
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 14
<400> 62
ccaaaacccc atgataagac 20
<210> 63
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 15
<400> 63
acatggcagc acacatacat 20
<210> 64
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 16
<400> 64
ggtcaggtaa ctgcacccta 20
<210> 65
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 17
<400> 65
gacatggcag cacacatac 19
<210> 66
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 18
<400> 66
ctaagaacag gcaggtctcc 20
<210> 67
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 19
<400> 67
tcagcctaag ctgggactac 20
<210> 68
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 20
<400> 68
gcttggcagg tttagaagac 20
<210> 69
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 21
<400> 69
gaatttctcc tggtccacag 20
<210> 70
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 22
<400> 70
acttcaggtt cagtccatgc 20
<210> 71
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 23
<400> 71
gcagaagata tagtgggcga 20
<210> 72
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 24
<400> 72
tcagaaaaga gctttggctt 20
<210> 73
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 25
<400> 73
caaaaagcca gcaactgtta 20
<210> 74
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 26
<400> 74
gatcagaaga tcccaaaagg 20
<210> 75
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 27
<400> 75
ctgtgataaa ggacgcgatt 20
<210> 76
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 28
<400> 76
aaggacaggg tgttcagaaa 20
<210> 77
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 29
<400> 77
gaaagacaga gagactgggc 20
<210> 78
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 30
<400> 78
gtggaaaacc aggtaagcac 20
<210> 79
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 31
<400> 79
tcagtctggg tggaaggtat 20
<210> 80
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 32
<400> 80
gagattttcc atgcagaagg 20
<210> 81
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 33
<400> 81
tgacctgctg tctcaaccta 20
<210> 82
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 34
<400> 82
gcaaccacat aacacacaga a 21
<210> 83
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 35
<400> 83
attggcagat tgtcggttac 20
<210> 84
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 36
<400> 84
acacattttg agggaggtgt 20
<210> 85
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 37
<400> 85
cattgtcatg tatcccacca 20
<210> 86
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 38
<400> 86
tgggagtcac ataatcatcg 20
<210> 87
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 39
<400> 87
tggctgtgta gagagcattg 20
<210> 88
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 40
<400> 88
ccgttttcat tcaagatacc 20
<210> 89
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 41
<400> 89
atcaagagaa tgggtgcagt 20
<210> 90
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 42
<400> 90
attatgtcct gtccaatgcc 20
<210> 91
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 43
<400> 91
accacgtcct tgagaaaaag 20
<210> 92
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 44
<400> 92
acaaactgac agggagagga 20
<210> 93
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 45
<400> 93
tggatttact gcaacattgg 20
<210> 94
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 46
<400> 94
gacaggtttt tcgcttttgt 20
<210> 95
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 47
<400> 95
cttaagggga cccacaagta 20
<210> 96
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223>downstream primer 48
<400> 96
cttgccttct gctagttttt g 21
<210> 97
<211> 154
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 1
<400> 97
accatctaga taaacaatgt atttgtccaa tctcaagcat tgttgggtca ttacagggtt 60
tccccaccct tgtattgatg gctctattgt tacctctgac tcccagttgt ttaataaacc 120
atattggtta cataaggcac agggtcataa caat 154
<210> 98
<211> 269
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 2
<400> 98
aactgactgc tcttgatggg taggaatgat caatcacagt taagtgaata ggagcctgac 60
tctaatataa ttttataagt gcagagccaa tgctagatga aattgaggag gcattgacgt 120
tgtttttgaa ctgaccacat cattcctctt ttctatgtcc ttctatgtca cgagcaatta 180
agcgactcag aggaagaaaa cgatgaaata gatggagtta atcatcaaca tttaccagcc 240
cgacgagccg aaccacaacg tcacacaat 269
<210> 99
<211> 263
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 3
<400> 99
tagacgacga tttcttacta aagacaagtt tggaagctct agagaagacc atccctaagc 60
ttttccaatt ctacttttct gattcagtgc ttgcaatata gaaaatgtaa atccacaatg 120
taccatagca caattaaaag acttgttaaa agtaaacaat aaacaaggag ctatgttagc 180
agtatttaaa gacacatatg ggctatcatt tacagattta gttagaaatt ttaaaagtga 240
taaaaccacg tgtacagatt gga 263
<210> 100
<211> 247
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 4
<400> 100
gtgcatgtag atttttattc tagaattaga gaattaagac attattcaga ctctgtgtat 60
ggagacacat tggaaaaact aactaacact gggttataca attttctaag aattaaacta 120
gatagtaaag ttcatctgcg ccacatttta gaaattgaaa gtacattgtg ggtgagactc 180
ttaagtttct taccatcact ttaactggga gaacctggtt ttatctatgt cttatctggg 240
gttttgg 247
<210> 101
<211> 224
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 5
<400> 101
gtctttcgag ctcagcagta gcacaatggc tgatccagaa ggtacagacg gggaggggct 60
ctgtttatat gctggattac atttattgat ttgcatatat tgaaccagcc ttgcatccca 120
gggatgaagc ccacttgatc atggtggata agcattttga tgtgctgctg gattcatttg 180
gccagtattt tattgaggat ttttgcatca atgttcatca agga 224
<210> 102
<211> 255
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 6
<400> 102
tgcatctgta cggctggttt atgtacagct attgtagaca aaaaaacagg agatgtaata 60
tcagatgacg aggacgaaaa tgcaacagac acagggtcgg atatggtaga ttttattgat 120
acacaaggaa cattttgtga acaggcagct ttctagatca aagagttcgg gagcgtgcct 180
gggtggagtg tgggaaccct ctggatgttt gctggaacgc tgctccacac tgtgtacctg 240
ggagggtaat ctttc 255
<210> 103
<211> 187
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 7
<400> 103
taagttacgc gagagtcaca tgatgcacaa gtgttgcatg ttttaaaacg aaagtttttc 60
atctgtcgca cagaaaactg ggggttgcaa agggagtagc ctctggttct tttgttactt 120
gggcatggaa agttgaggtt ttccttttgc tttagttcta ggaagtaagt ttgaatcagc 180
ctgagga 187
<210> 104
<211> 247
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 8
<400> 104
gagatttacg agtttgcagg aggcagcaag aaacagtcca ttaggggagc tgtaggaatg 60
aggttcccat tttgtggctc cctgcaagcc agggactatt ctcagtttct ggagactact 120
tcagttcctt gccatgcagc ttcctcttaa gtgcagcttc tcacatttca aatctatcta 180
actccaggaa gggcccagtc ccttataaag ctcactgatt atgtcaagcc caccttgata 240
atcactc 247
<210> 105
<211> 271
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 9
<400> 105
acgtagtatc acatgtacca tagcacaatt aaaagacttg ttaaaagtaa acaataaaca 60
aggagctatg ttagcagtat ttaaagacac tgtgtagagc cacttgcttt atgaatctgg 120
gtgctcctgt attgggtgca tatatattta ggatagttag ctcttcttgt tgaattgatc 180
cctttaccat tatgtaatgg ctttctttgt ctcttttgat ctttgttggt ttaaagtctg 240
ttttatcaga gactaggatt gcaacccctc a 271
<210> 106
<211> 235
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 10
<400> 106
actaataagg ccatttatat tatatgccca tattcaatgt ctagactgta aatggggagt 60
attaatatta gccctgttgc gttacaaatg tggtaagagt agactaacag ttgctaaagg 120
tttaagtacg ttgttacacc ctgcaactag tgcctgccaa tgagtgagaa agagtcagca 180
ctgccgttga ctgcatgttg actgtgcacc aggcattgtc cctgcatttt tctca 235
<210> 107
<211> 259
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 11
<400> 107
ccaatacagt acggtatatc aatattagtg aagtaatggg agacacacct gagtggatac 60
aaagacttac tattatacaa catggaatag atgatagcaa ttttgatttg tcagtactag 120
tctagaatct caaggtgttg gcagagagcc tgccccataa tggacaatca acatgcaccg 180
gctcactgaa tgggcacaaa gatgtccctt ccattttgtt ctcaccgatc tctgggtcct 240
agcacagacc tggaactta 259
<210> 108
<211> 150
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 12
<400> 108
catgctagtg cattctgcga taccaacaaa tagagtttat aacattttta ggagccttag 60
catctgagaa gtcatccatt gctcgggaaa aaaacttgca ataatatcag ttctaatacc 120
aatgccttct aaacgtatca atcaggcacc 150
<210> 109
<211> 135
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 13
<400> 109
gtatggatct gaattaatta ccatctacta cctctgtatg gcccattcat ataataaata 60
ttgctttgtg ttctatctgg gcatggtatt agcaatatat gttagatgaa gcagggattt 120
gggagtcagt tgtca 135
<210> 110
<211> 220
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 14
<400> 110
cggacgtatt agcagacaca gttatgtatt ttgggctgtg cccctgctat tggggaacac 60
tgggctaaag gcactgcttg taaatcgcgt cctttatcac agggcgattg cgccacattt 120
tagaaattga aagtacattg tgggtgagac tcttaagttt cttaccatca ctttaactgg 180
gagaacctgg ttttatctat gtcttatcat ggggttttgg 220
<210> 111
<211> 242
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 15
<400> 111
ggtaataaac gcgcagctag ttttgtattt ttagtggaga cggggtttca ccatgttggt 60
caggctggtc ttgaactcct gacctcatga tcctctcacc tcggcctcac aaagtgctag 120
gattacaggc cacctggcat tggacaggtg caggcaatga aaaaacagga atactgactg 180
taacatacca tagtgaaaca caaagaacaa aatttttaaa tactgttgca attccagata 240
ga 242
<210> 112
<211> 253
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 16
<400> 112
ggactctaga taacttcatc actacctctg actggtcagt attaagaacc actttcaggc 60
caggtgtcat ggttcacgcc tgtaactcca tctactccag aggctgaggc aggacaattg 120
cttcaggccg gaggattatt attaactgtt ggtaatccat attttagggt tcctgcaggt 180
ggtggcaata agcaggatat tcctaaggtt tctgcatacc aatatagagt atttagggtg 240
cagttacctg acc 253
<210> 113
<211> 249
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 17
<400> 113
acggggcaag atcagcgcgg cctttcccag ccttagcgag gcgccctgca gcctggtacg 60
cgcgtggcgt ggcggtgggc gcgcagtgcg ttctcggtgt ggagggcagc tgttccgcct 120
gcgatgattt atactcacag gacaaggatg cggtttgtca aacagtactg ctacggagga 180
gtaccaatat gttatcactt atttttttat tttgcttttg tgtatgcatg tatgtgtgct 240
gccatgtca 249
<210> 114
<211> 221
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 18
<400> 114
aattaaatca gccatttata ttatatgccc atattcaatg tctagactgt aaatggggag 60
tattaatatt aggccaagaa tgaatgatta tttgtgcctt cctatgtgaa aaaaaaatgt 120
ttcctctagc tacacactat tctgttctgt gaggccgccc catcagactg ttgacctaga 180
gtcccaaccc cggccctcca ggagacctgc ctgttcttag a 221
<210> 115
<211> 217
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 19
<400> 115
ccccgactgt gtcatatgtt atcacttatt tttttatttt gcttttgtgt atgcatgtat 60
gtgtgctgcc atgtctctac taaacacaca cacacacaca cacacaaatt agctggccgt 120
ggtgacagcc tgtagtccca gcttaggctg aacacaaaca cacacacaca aattagctgg 180
ccgtggtgac agcctgtagt cccagcttag gctgaaa 217
<210> 116
<211> 234
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 20
<400> 116
cccgcctatg cctcgttctt ttttgatgcc tatgtagtac ttatacatct atcgcatctt 60
tcatctcaaa tcctcatggc attccccctc tgtttgttat cattacaccc agaatggctc 120
tttgaggtct cacagtcatt ttcttggttc aggctggatt gcgtcgcaag cccaccatag 180
gccctcgcaa acgttctgct ccatctgcca ctacgtcttc taaacctgcc aagc 234
<210> 117
<211> 184
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 21
<400> 117
ccgctgcgat agaggagaac gatgaataga tggagttaat catcaacatt taccagcccg 60
acgagccgaa ccacaacgtc acacaatgtt gtgtatgttt gttatcatta cacccagaat 120
ggctctttga ggtctcacag tcattttctt caaggtggag aaactgtgga ccaggagaaa 180
ttca 184
<210> 118
<211> 190
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 22
<400> 118
cctagatatt agtatgtatt tgttggtacc cctacatctg gaacacatgg gtatgaggaa 60
atacctttac aaacatttgc ttcttctggt acgggggagg aacactggag ccatctcagc 120
ctaatggcct tccaattaaa tgttttttaa aagttttaag actcaaattt ggcatggact 180
gaacctgaag 190
<210> 119
<211> 230
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 23
<400> 119
ggtagctcaa cactcttcat tgataactat agtgtcattc tggtaattat cattaatttt 60
atttaccaga atattggtga tgtctgcaga caggagaatt agtggggtat cttgtttcgg 120
ttctggagtt tgatatatat gcagatgaca tggaccctgc agtgcctgta ccatcgcgtt 180
ctactacctc ctttgcattt tctaaatatt cgcccactat atcttctgca 230
<210> 120
<211> 240
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 24
<400> 120
agctatagtt atacacatgg atagatgata gcaattttga tttgtcagaa atggtacaat 60
gtttttacat gcattttctt atttgagcca cataataaac ctctttacat ccttgttata 120
attgagaaat ctgaaattca aatactgcaa gtaattcatt tgatattcct taattaataa 180
ttggtagacc tggtattcag attaggaact tctgaattca aagccaaact cttttctgaa 240
<210> 121
<211> 151
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 25
<400> 121
ttaagggata gtgcattcct gcgataccaa caaatagagt ttataacatt ttctgaatat 60
tctggactag gggatatctc agaacagtac ttagcaccta gtgtaccccc catcaggttt 120
cccttatagt taacagttgc tggctttttg a 151
<210> 122
<211> 194
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 26
<400> 122
tatggaggag ctgttcgctt tagtagatta ggtcaagggc aactatgttg ctcatgaagt 60
atcgttgctt ggatctcctc ttttggagac taagggagct tgactgtgtg caagttagca 120
ataaggactg actactggat taggtggacc acgtgaatat catcttgtgg tggccttttg 180
ggatcttctg atca 194
<210> 123
<211> 237
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 27
<400> 123
atgacttctg atatgggagc cctggcttcc agtaatagaa agatcaatat gcagaagcta 60
cattgtaagt tgcatgagaa tggcagggtc tgtcttgctc cagttatatt cccagtgtta 120
gggacaatgt gtctgtagat tataagcaga cacagttatg tattttgggc tgtgcccctg 180
ctattgggga acactgggct aaaggcactg cttgtaaatc gcgtccttta tcacaga 237
<210> 124
<211> 261
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 28
<400> 124
ttgtctatct agaagtgttg ttcataagta taaagtttca gttatgctag attaataaat 60
tctagaggtc ttctatacaa catattatct actgttaata atatgatatt gtgcaccaga 120
aaatttaaga gggaagatct cttaaacata caatgttgtg tatgtgttgt aagtgtgaag 180
ccagaattga gctagtagta gaaagctcag cagacgacct tcgagcattc cagcagctgt 240
ttctgaaaac ctgtcctaag a 261
<210> 125
<211> 237
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 29
<400> 125
ggccgtcttt acgcattgcc agctaaatat ttaaaagatt gtgccacaat gtgcaaacat 60
tataggcgag cccaaaaacg acaaatgaat atgtcacagt gtttgagtaa tcatatataa 120
acataaataa ctataaaata tgcacacaca gaaaaagata gtacagaagt agtgttaaga 180
tatagcatta ttccaagatt taaataatgt tacaattgcc cagtctctct gtctttc 237
<210> 126
<211> 277
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 30
<400> 126
tgtggaaaac ccagtaagca ccgaagtcca cttgcctttt aatttatttt tttatcactt 60
taatgctgag atgagtcgaa tgcctagatg aggtggtggg tgtagctttt cgttttccta 120
atgtaaattt tggtttggcc tttaatcctg cttgtagtaa aaatttgcgt cctaaaggaa 180
actgatctag gtctgcagaa aacttttcct ttaaatttac ttcccaaaaa gtgtattttt 240
taaggggatc ttctttaggt gctggaggtg tatgtta 277
<210> 127
<211> 213
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 31
<400> 127
ttcagtctgg gtggaaggta tccaatccag atagctgtgc atacataatg cataatacat 60
gactcccccc tttcccaacg tattagttgc caatactgta tttggctgtc tatgtcttta 120
ctgtcatttt catagtggtc tatgattttg tcctgcaacg cacttaaacg ttccgaaagg 180
gtttccttcg gtgtctgcat ttcctcttcc tca 213
<210> 128
<211> 290
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 32
<400> 128
ttttgagatt ttcccagcag aagggactgg gcatcaactg tcaggacatg gacagagacg 60
tctgccatta gaagctggtc caacacctaa aggctgacca cggccaattt ccactccagc 120
acaggcccac actaaacgtt gtgtttcagg attataaata ctagtatcag gtaaaccaaa 180
tttatttggg tcaggtaact gcaccctaaa tactctatat tggtatgcag aaaccttagg 240
aatatcctgc ttattgccac cacctgcagg aacctaaaaa tatggagagg 290
<210> 129
<211> 236
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 33
<400> 129
ttccaaccta ttcctgcatg tctgtgtcct ctgcgtaccg tctgtctcct gtctggacct 60
cagctttttc ccctgtaaaa tgaggctgtc cactcacgtg acattagtag tctttccatc 120
cagattgttc attaggaaat gtaaacacca ccaatctatt atgtaaatac ggccacctag 180
aatctgtacc agcattaatg ttagatgtaa ttaataatgg agggcatttt agttga 236
<210> 130
<211> 257
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 34
<400> 130
atcaaagtct ggaagggtgc agagcctgtc ctggaatgga gggaacaggt gaagcgccgg 60
actctcctgg ggataatatg tgtgtgtgta tatatatata catctattgt tgtgtttgta 120
tgtcctgtgt ttgtgtttgt tgtatgattg cattgtatgg tatgtatggt tgttgttgta 180
tgttgtatgt tactatattt gttggtatgt ggcattaaat aaaatatgtt ttgtggttct 240
gtgtgttatg tggttgc 257
<210> 131
<211> 255
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 35
<400> 131
aaggatatgc gagaggtcac atgatgcaca agtgttgcat gttttaaaac gaaagtttgc 60
aggaggcagc acagaaaaca gtccattagg ggagcggctg gaggtggata cagagttaag 120
tccacggtta caagaaatat ctttaaatag tgggcagaaa aaggcaaaaa ggcggctgtt 180
tacaatcccc agccatatgg cgcaagaaca ctaatacagt tgtctgtgca caactgtaac 240
cgacaatctg ccaat 255
<210> 132
<211> 227
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 36
<400> 132
agtagatctt ttcaaccatc ttttcaaatg ccatatcgct ttcatctgtc agctcattat 60
caaatgccca ttgtaccatt tctgacaaat caaaattgct atcatctatt ccatgttgta 120
taatagtaag tctttgtatc cactcaggtg tgtctcccat tacttcacta atatttgata 180
ttcctgttct ataccaatat agtgctgcaa cacaccttcg caataaa 227
<210> 133
<211> 254
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 37
<400> 133
ccaactactt gctttatata tttttctagg gaaaaaatac cagtgttata gtactataca 60
atgcaaataa aataaaatat caataaacta aatctaaaat ttgcatttag gattctatta 120
gataaattga ggacaggtgc aggcaatgaa aaaacaggaa tactgactgt aacataccat 180
agtgaaacac aaagaacaaa atttttaaat actgttgcaa ttccagatag tgtacaaata 240
ttggtgggat acat 254
<210> 134
<211> 192
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 38
<400> 134
cctatgtgag aatgaggtga gatctttata atgatacatt tccatttaat gaatagtaaa 60
tatattttct gttctttatg attttcccta tttttttgca gatggctttg tggcggccta 120
gtgacaatac cgtatatctt ccacctcctt ctgtggcaag agttgtaaat accgatgatt 180
atgtgactcc ca 192
<210> 135
<211> 266
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 39
<400> 135
gcgtctcaat ggcgactggc ggcatgtatg tagtggcggc agtacggagg ctatagacaa 60
cgggggcaca gagggcaaca acagcagtgt aatcaccatt ctttctatct ccatgaacct 120
actctaggca cttcgtgcaa gtgcaatcat atgatatttt tccttttgtg tgactagatt 180
atttcactta gcatactgac ttcaaagttc tgacacatgc tgtagcatgt gtcacaattt 240
tttttcaatg ctctctacac agccaa 266
<210> 136
<211> 263
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 40
<400> 136
gcatgatatg cgttagatgg catccaataa gtattgatag aaagcacaaa ccattaatac 60
aactaaaatg tcctccaata ctactaacca caaatataca tccagcaaag gataatagat 120
ggccatattt agaaagtaga ataacagtat ttgaatttcc aaatgcattt ccatttgact 180
gtacttaagt acagtccctc caaaatttcc ataaacaaat ttctaagaaa tatgagctaa 240
caaggtatct tgaatgaaaa cgg 263
<210> 137
<211> 175
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 41
<400> 137
ggactgtagt gtacctacgc ctataataca tttaaaaggt gacagaaaca gtttaaaatg 60
tttacggtac agattgcgaa aacatagcga ccactataga cctatctgca acaataaaaa 120
gcaggaagac ttagtccctt cacagattgt tatcactgca cccattctct tgata 175
<210> 138
<211> 254
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 42
<400> 138
gatttcgctt tcgttcgcag acactttgaa gtcttgttaa agggcaatac ttgtgggtcc 60
ccttaagatg tgtctctgtg tgtgtgtgtg tgtgtatgtg tgtgtgtgtt acatttgtct 120
gaactgcaac aaccaagcat ggccaaacat ttaaaaggtg atgctaatac tttaaaatgt 180
ttaagatata tatttaaaaa gcattgtaaa ttgtatactg cagtgtcgtc tacatggcat 240
tggaaggaca taat 254
<210> 139
<211> 256
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 43
<400> 139
ccagtcgcat atgtcaaacc agaatttcag gcaacttagt aacaggacac gctaccaaca 60
ccattggtac aactaaaatg ccctccatta ttaattacat ctaacattaa tgctggtaca 120
gattctaggt ggccttattt acataataga ttggtggtgt ttacatttcc taatgagttt 180
ccatttgacg aaaacggaaa tccagtgtat gagcttaatg ataagaactg gaaatccttt 240
ttctcaggac gtggta 256
<210> 140
<211> 256
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 44
<400> 140
cctcgagcgt aaggttctaa cgaaagtatt tgggtagtcc acttagtgat attagtggat 60
gtgtagacaa taatattagt cctagattaa aagctatatg tatagaaaaa caaagtagag 120
ctgcaaaaag gagattattt gaaagcgaag acccttactc aacaataccc gggaaacatc 180
gcgcgctggc cagccgcgga gacacagttc ccaagcaacg cataaactga agaacttcct 240
ctccctgtca gtttgt 256
<210> 141
<211> 294
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 45
<400> 141
cttttccctg gaaataactg agcactttga atttttttta tgtctaaaat tgtcagttaa 60
tttattattt tgtttgagta agaattttaa tattgccata ttctgtagta tttttctttg 120
tatatttcta gtatggcaca tgatatgagt cactggataa tagagaatgt atatctatgg 180
attacaaaca aacacaattg tgtttaattg gttgcaaacc acctataggg gaacactggg 240
gcaaaggatc cccatgtacc aatgttgcag taaatccaag gtgggggggg gttg 294
<210> 142
<211> 238
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 46
<400> 142
gaatttgaca ggtttttcgc ttttgttttc ggttgaggac ttactacccc aaaaacacat 60
ccgacctgta ctagaatagg tgtgtgaggg agcgggaaaa gctcgggtca ttttaactgg 120
actagaaagg aacacagcga tcctttttgt catatagaca taaatccagt agacactgta 180
atagtttttg gtattttaac ttgagacaaa aattgttcac gttgccattc actatcaa 238
<210> 143
<211> 252
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 47
<400> 143
ttttttctta aggggaccca caagtattgc cctttaacaa gacttcaaag ttttctgctg 60
taaagaaagc tgtaatatat agtaaatata tagtaaaact aaatgttgcg tgtatatcca 120
tgttttttta tacatcctgt tggtgcagtt aaatacactt caaggctaac gtcttgtaat 180
gtccactttt cattactata ttgtgagtta tatattgttt ctaacgttag ttgcagttca 240
attgcttgta aa 252
<210> 144
<211> 243
<212> DNA
<213> Artificial Sequence
<220>
<223>integration sequence 48
<400> 144
tcttgccttc tgctagtttt tgaatgtgtt tgctcttgct tctctagttc ttttaattgt 60
gttgttaggg tggttaatat gtttaaatcc catttctctg gccttgtaat aaatagcaca 120
ttctaggcgc atgtgtttcc aatagtctat atggtcacgt aggtctgtac tatcattttc 180
ataatgtgtt agtattttgt cctgacacac atttaaacgt tggcaaagag tctccatcgt 240
tta 243
Claims (10)
1. a kind of for diagnosing the biomarker of cervical cell smearing, which is characterized in that the biomarker is following base
Because of family: MYC gene family, NR4A gene family or ANKRD gene family or combinations thereof or its encoded albumen, wherein
The biomarker is integrated with the partial sequence or full sequence of HPV.
2. biomarker as described in claim 1, which is characterized in that the MYC gene family includes MYC gene and MYCN
Gene；The NR4A gene family includes NR4A2 gene；The ANKRD gene family includes ANKD12 gene.
3. biomarker as claimed in claim 1 or 2, which is characterized in that the HPV is HPV16 and/or HPV18.
4. biomarker as described in any one of claims 1-3, which is characterized in that the biomarker integrates HPV's
Partial sequence or full sequence are as shown in SEQ ID NO:97 to SEQ ID NO:144.
5. biomarker according to any one of claims 1-4, which is characterized in that the biomarker integrates HPV's
Partial sequence or full sequence generate breakpoint, and the breakpoint includes the breakpoint such as below table on HPV breakpoint and human genome
It is shown.
6. biomarker as described in any one in claim 1-5, which is characterized in that the biomarker further includes
INSM1 and/or ASCL1.
7. biomarker as claimed in any one of claims 1 to 6, which is characterized in that the biomarker by WGS,
VCS or immunohistochemistry detection.
8. if the described in any item biomarkers of right 1-7 are in the diagnosis marker for preparing cervical cell smearing or containing its examination
Purposes in agent box.
9. a kind of kit for diagnosing cervical cell smearing, which is characterized in that the kit includes for detecting as right is wanted
Seek the reagent of any one of 1-7 biomarker；Preferably, the reagent is primer, it is highly preferred that the amplification of the primer
Product is the sequence as shown in SEQ ID NO:97 to SEQ ID NO:144；
Most preferably, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:1, downstream nucleotide sequence such as SEQ ID
Shown in NO:49；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:2, downstream nucleotide sequence such as SEQ ID
Shown in NO:50；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:3, downstream nucleotide sequence such as SEQ ID
Shown in NO:51；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:4, downstream nucleotide sequence such as SEQ ID
Shown in NO:52；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:5, downstream nucleotide sequence such as SEQ ID
Shown in NO:53；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:6, downstream nucleotide sequence such as SEQ ID
Shown in NO:54；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:7, downstream nucleotide sequence such as SEQ ID
Shown in NO:55；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:8, downstream nucleotide sequence such as SEQ ID
Shown in NO:56；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:9, downstream nucleotide sequence such as SEQ ID
Shown in NO:57；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:10, downstream nucleotide sequence such as SEQ
Shown in ID NO:58；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:11, downstream nucleotide sequence is such as
Shown in SEQ ID NO:59；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:12, downstream nucleotide sequence
As shown in SEQ ID NO:60；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:13, downstream nucleotide sequence
Column are as shown in SEQ ID NO:61；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:14, downstream nucleotide
Sequence is as shown in SEQ ID NO:62；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:15, downstream nucleotide
Acid sequence is as shown in SEQ ID NO:63；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:16, downstream core
Nucleotide sequence is as shown in SEQ ID NO:64；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:17, downstream
Nucleotide sequence is as shown in SEQ ID NO:65；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:18, under
Nucleotide sequence is swum as shown in SEQ ID NO:66；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:19,
Downstream nucleotide sequence is as shown in SEQ ID NO:67；Or, the upstream nucleotide sequence of the primer such as SEQ ID NO:20 institute
Show, downstream nucleotide sequence is as shown in SEQ ID NO:68；Or, the upstream nucleotide sequence of the primer such as SEQ ID NO:21
Shown, downstream nucleotide sequence is as shown in SEQ ID NO:69；Or, the upstream nucleotide sequence of the primer such as SEQ ID NO:
Shown in 22, downstream nucleotide sequence is as shown in SEQ ID NO:70；Or, the upstream nucleotide sequence of the primer such as SEQ ID
Shown in NO:23, downstream nucleotide sequence is as shown in SEQ ID NO:71；Or, the upstream nucleotide sequence of the primer such as SEQ
Shown in ID NO:24, downstream nucleotide sequence is as shown in SEQ ID NO:72；Or, the upstream nucleotide sequence of the primer is such as
Shown in SEQ ID NO:25, downstream nucleotide sequence is as shown in SEQ ID NO:73；Or, the upstream nucleotide sequence of the primer
As shown in SEQ ID NO:26, downstream nucleotide sequence is as shown in SEQ ID NO:74；Or, the nucleotides upstream sequence of the primer
Column are as shown in SEQ ID NO:27, and downstream nucleotide sequence is as shown in SEQ ID NO:75；Or, the nucleotides upstream of the primer
Sequence is as shown in SEQ ID NO:28, and downstream nucleotide sequence is as shown in SEQ ID NO:76；Or, the upstream nucleosides of the primer
Acid sequence is as shown in SEQ ID NO:29, and downstream nucleotide sequence is as shown in SEQ ID NO:77；Or, the upstream core of the primer
Nucleotide sequence is as shown in SEQ ID NO:30, and downstream nucleotide sequence is as shown in SEQ ID NO:78；Or, the upstream of the primer
Nucleotide sequence is as shown in SEQ ID NO:31, and downstream nucleotide sequence is as shown in SEQ ID NO:79；Or, the primer is upper
Nucleotide sequence is swum as shown in SEQ ID NO:32, downstream nucleotide sequence is as shown in SEQ ID NO:80；Or, the primer
Upstream nucleotide sequence is as shown in SEQ ID NO:33, and downstream nucleotide sequence is as shown in SEQ ID NO:81；Or, the primer
Upstream nucleotide sequence as shown in SEQ ID NO:34, downstream nucleotide sequence is as shown in SEQ ID NO:82；Or, described draw
The upstream nucleotide sequence of object is as shown in SEQ ID NO:35, and downstream nucleotide sequence is as shown in SEQ ID NO:83；Or, described
The upstream nucleotide sequence of primer is as shown in SEQ ID NO:36, and downstream nucleotide sequence is as shown in SEQ ID NO:84；Or, institute
The upstream nucleotide sequence of primer is stated as shown in SEQ ID NO:37, downstream nucleotide sequence is as shown in SEQ ID NO:85；Or,
The upstream nucleotide sequence of the primer is as shown in SEQ ID NO:38, and downstream nucleotide sequence is as shown in SEQ ID NO:86；
Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:39, downstream nucleotide sequence such as SEQ ID NO:87 institute
Show；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:40, downstream nucleotide sequence such as SEQ ID NO:88
It is shown；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:41, downstream nucleotide sequence such as SEQ ID NO:
Shown in 89；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:42, downstream nucleotide sequence such as SEQ ID
Shown in NO:90；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:43, downstream nucleotide sequence such as SEQ
Shown in ID NO:91；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:44, downstream nucleotide sequence is such as
Shown in SEQ ID NO:92；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:45, downstream nucleotide sequence
As shown in SEQ ID NO:93；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:46, downstream nucleotide sequence
Column are as shown in SEQ ID NO:94；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:47, downstream nucleotide
Sequence is as shown in SEQ ID NO:95；Or, the upstream nucleotide sequence of the primer is as shown in SEQ ID NO:48, downstream nucleotide
Acid sequence is as shown in SEQ ID NO:96.
10. kit according to claim 9, which is characterized in that further include that DNA extracts reagent and PCR amplification reagent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811519133.4A CN109355393A (en) | 2018-12-12 | 2018-12-12 | Cervical cell smearing relevant HPV integrator gene site and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811519133.4A CN109355393A (en) | 2018-12-12 | 2018-12-12 | Cervical cell smearing relevant HPV integrator gene site and its application |
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| Publication Number | Publication Date |
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| CN109355393A true CN109355393A (en) | 2019-02-19 |
Family
ID=65328643
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811519133.4A Pending CN109355393A (en) | 2018-12-12 | 2018-12-12 | Cervical cell smearing relevant HPV integrator gene site and its application |
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| Country | Link |
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| CN (1) | CN109355393A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110229895A (en) * | 2019-06-04 | 2019-09-13 | 广州鼓润医疗科技有限公司 | A kind of molecular marked compound of uterine neck intraepithelial neoplasia that predicting human papilloma virus positive patient |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107002136A (en) * | 2014-09-22 | 2017-08-01 | 格罗宁根大学 | The biomarker of cervical carcinoma |
| CN107846903A (en) * | 2015-05-28 | 2018-03-27 | 细胞结构公司 | The stem cell in placenta source and its for recovering regeneration engine, correcting protein group defect and extending the purposes in aging individuals life-span |
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