CN109136080A - A kind of detection kit and its application for the early diagnosis of straight colon cancer - Google Patents
A kind of detection kit and its application for the early diagnosis of straight colon cancer Download PDFInfo
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- CN109136080A CN109136080A CN201811112982.8A CN201811112982A CN109136080A CN 109136080 A CN109136080 A CN 109136080A CN 201811112982 A CN201811112982 A CN 201811112982A CN 109136080 A CN109136080 A CN 109136080A
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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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- 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
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Abstract
It is disclosed by the invention to belong to detection kit technical field, specially a kind of detection kit and its application for the early diagnosis of straight colon cancer, including box body and box cover, the box cover is screwed onto the upside of box body, the bottom of the box body is provided with reflector, filler is provided on the right side of the top of the box cover, round through slot is offered in the middle part of the ontology of the box cover, and photoscope has been laterally embedded into round through slot, storage of the detection kit for straight colon cancer early diagnosis reagent, pass through the setting of reflector, it can be improved the intensity of reflected light, convenient for in detection kit medicament and test object observe；By the setting of filler, reduces and open and the step of closed box, reduce the pollution of extraneous bacterium, guarantee the accuracy detected；By the setting of rubber stopper, conical plate, internal reagent outflow can be avoided；It, can be through the reagent situation of photoscope observation tray interior by the setting of photoscope；Structure is simple, easy to use.
Description
Technical field
The present invention relates to detection kit technical field, specially a kind of detection reagent for the early diagnosis of straight colon cancer
Box and its application.
Background technique
Due to the change of living habit, the disease incidence of China's its people's colorectal cancer increases year by year, has been in each tumour of men and women
First 3~4 of disease incidence.Operative treatment can be such that early stage colorectal cancer is eradicated, and advanced colorectal cancer is combined through chemoradiotherapy
The mean survival time is still less than 30 months after molecular targeted therapy.Therefore, the early diagnosis of colorectal cancer seem it is extremely important and
Urgently.The fiber Colon and rectum mirror that colorectal cancer early diagnoses goldstandard is a kind of invasive inspection, it is desirable that fully and completely enteron aisle
Prepare, the inconveniences such as exposure and calmness of privacy, it is difficult to be accepted extensively by people.Fecal occult blood testing is current clinical recommendation
Colorectal cancer early screening standard non-intruding means, however its sensibility is poor, differs from 15%~80%, most document
For report less than 30%, specificity is relatively low, is not able to satisfy the early diagnosis of colorectal cancer.Recent study shows DNA methylation
State change is in close relations with the occurrence and development of colorectal cancer, and crosses tumor development process through entire；It has found that
The abnormal hyper-methylation of multiple genes frequently occurs in colorectal cancer.Therefore, the exception of DNA methylation is expected to as Colon and rectum
The molecular marker of cancer early diagnosis.There is free tumour correlation DNA in Peripheral Blood from Patients with Malignant and has caused oncology
The very big concern on boundary detects one of the hot spot that tumor related marker object in blood of cancer patients is current cancer research.Research
Personnel have found DNA variation identical with primary tumo(u)r in kinds of tumors peripheral blood in patients.Therefore, this research is intended to pass through detection
Biology of the DNA gene methylation as a kind of straight colorectal cancer clinical diagnostic means of atraumatic in straight peripheral blood of patients with colonic cancer
Index, the current country there is no the relevant report of this aspect.
Colon cancer is the common malignant tumor of digestive tract for betiding colon site, is apt to occur in rectum and sigmoid colon has a common boundary
Place, with 40~50 years old age group disease incidence highest.Disease incidence accounts for the 3rd of gastroenteric tumor.Colon cancer is mainly gland cancer, mucus
Gland cancer, undifferentiated carcinoma.General form is in polypoid, ulcer type etc..Based on performing the operation, auxiliary is given chemotherapy and is exempted from for the treatment of colon cancer
Epidemic disease treatment etc..Therefore early to be the discovery that the key for effectively being treated and being extended patient vitals.Currently, the diagnosis of colon cancer mainly according to
By the use of Sigmoidoscope.It is well known that Sigmoidoscope is brought great pain to patients.In terms of molecular diagnosis and prognosis judgement still
Without accurate, sensitive, special method.
Free serum DNA is the research hotspot of many tumours and other associated biomolecule fields now.It is a large amount of clinical and face
Bed early period the experimental results showed that containing a large amount of or a certain amount of dissociative DNA in the serum of tumour or tumour early stage patient.These
Free serum DNA is mainly from tumour cell downright bad, apoptosis in blood circulation；Also it is possible that directly being discharged by tumour cell
Into blood.These free serum DNAs have become the research hotspot of modern oncology, be tumour early diagnosis, prevention and
The follow-up for the treatment of provides reliable foundation and new direction.The methylation of DNA tumour generation, develop and lapse to process hair
Wave important role.As an a kind of important branch of epigenetics, the study of pathogenesis of the methylation of DNA in tumour
And treatment method probe into and the early diagnosis of tumour in play an important role.This item development plan is using analysis early stage
The methylation of several gene promoter areas in the DNA to dissociate in colorectal cancer patients serum, in conjunction with modern biological information skill
Art and Principle of Statistics can early stage it is special, it is sensitive, accurately examine colon cancer, while being mentioned for the follow up and prognosis of the patient
For the foundation of science.Such as China Patent Publication No. be CN104561258B relate to it is a kind of for straight colon cancer early diagnosis
Detection kit, the detection kit include three pairs and detect rRNA_pseudogene, HECTD1 and ZNF843 gene respectively
Promoter and gene inner body zone methylation specificity amplification primer and a methylation-specific sequencing primer, described three pairs points
Not Jian Ce rRNA_pseudogene, HECTD1 and ZNF843 gene promoter zone methylation specificity amplification primer includes methyl
Change specific forward primer and methylation-specific downstream primer；It is by detecting rRNA_ relevant to straight colon cancer
The gene promoter area pseudogene, HECTD1 and ZNF843 and the straight colon cancer of gene inner body methylation auxiliary diagnosis, letter
Single, conveniently, detection efficiency is high, with strong points, has the advantages that accurate and reliable, flexible quick and economy, is conducive to directly tie
The early detection of intestinal cancer and in time treatment.But it is when in use, it is not convenient enough for the storage of detection reagent, observation, taking-up.
Summary of the invention
The purpose of the present invention is to provide a kind of detection kits and its application for the early diagnosis of straight colon cancer, with solution
Existing detection kit certainly mentioned above in the background art when in use, for the storage of detection reagent, observation, is taken out not
Enough convenient problems.
To achieve the above object, the invention provides the following technical scheme: a kind of detection for the early diagnosis of straight colon cancer
Kit, including box body and box cover, the box cover are screwed onto the upside of box body, and the bottom of the box body is provided with reflector, institute
It states and is provided with filler on the right side of the top of box cover, round through slot is offered in the middle part of the ontology of the box cover, and horizontal in round through slot
To photoscope is embedded with, the inner lower of the filler is embedded with rubber stopper.
Preferably, the box body and filler integrated injection molding.
Preferably, the bottom of the box body is provided with stabilizer blade, and stabilizer blade has been embedded in rubber pad, and rubber pad passes through bonding
Agent is Nian Jie with stabilizer blade.
Preferably, the reflector includes plastic protective plate and masking foil, and masking foil is placed in the downside of plastic protective plate, described
The edge of plastic protective plate and the intracavity bottom of box body are Nian Jie.
Preferably, the bottom of the filler is provided with conical plate, and conical plate is in hollow funnel-form, the conical plate
It is coaxially disposed with filler.
Preferably, lateral retaining ring is provided on the downside of the inner cavity of the filler, and lateral retaining ring is supported under rubber stopper
Side.
Preferably, storage of the detection kit for straight colon cancer early diagnosis reagent.
Compared with prior art, the beneficial effects of the present invention are: the detection kit for being used for straight colon cancer early diagnosis
Advantage be:
1) by the setting of reflector, can be improved the intensity of reflected light, convenient for in detection kit medicament and inspection
Object is surveyed to be observed；
2) it by the setting of filler, reduces and opens and the step of closed box, reduce the pollution of extraneous bacterium, guarantee to examine
The accuracy of survey；
3) setting for passing through rubber stopper, conical plate can be avoided internal reagent outflow；
It 4), can be through the reagent situation of photoscope observation tray interior by the setting of photoscope；
5) structure is simple, easy to use.
Detailed description of the invention
Fig. 1 is schematic structural view of the invention；
Fig. 2 is the schematic cross-sectional view of Fig. 1 of the present invention；
Fig. 3 is schematic structural view of the invention；
Fig. 4 is schematic structural view of the invention.
In figure: 1 box body, 2 reflectors, 3 box covers, 4 fillers, 5 photoscopes, 6 rubber stoppers.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
Fig. 1-3 is please referred to, the present invention provides a kind of technical solution: a kind of detection reagent for the early diagnosis of straight colon cancer
Box, including box body 1 and box cover 3, box cover 3 are screwed onto the upside of box body 1, and the bottom of box body 1 is provided with reflector 2, the top of box cover 3
End right side is provided with filler 4, round through slot is offered in the middle part of the ontology of box cover 3, and be laterally embedded into perspective in round through slot
Mirror 5, the inner lower of filler 4 are embedded with rubber stopper 6.
Wherein, box body 1 and 4 integrated injection molding of filler, leakproofness is preferable, and the bottom of box body 1 is provided with stabilizer blade, and props up
Foot has been embedded in rubber pad, so as to steadily put, and rubber pad is Nian Jie with stabilizer blade by bonding agent, reflector 2 includes
Plastic protective plate and masking foil, and masking foil is placed in the downside of plastic protective plate, increases reflecting effect, the edge of plastic protective plate and box
The intracavity bottom of body 1 is bonded, and avoids medicament from immersing on masking foil, the bottom of filler 4 is provided with conical plate, and conical plate is in
Hollow funnel-form, conical plate and filler 4 are coaxially disposed, and are reduced reagent outflow, are provided with transverse gear on the downside of the inner cavity of filler 4
Circle, and lateral retaining ring is supported on the downside of rubber stopper 6, plays the role of supporting rubber stopper 6.
Storage of the detection kit for straight colon cancer early diagnosis reagent.
Peripheral blood of patients with colorectal cancer is collected, to obtain early diagnosis information, only selecting clinical stages in rectum disease is I the or II phase
Patient, III or IV are excluded.Patient excludes have other outer tumours of the carcinoma of the rectum, takes out non-underwent operative or chemotherapy before peripheral blood.
Optionally learn from else's experience proctoscope and pathologic finding without tumor cases is control group.Laboratory is sent in 30min after periphery blood specimen is collected, is stood
I.e. extract genomic DNA, be placed in -20 DEG C freeze it is spare.The equal informed consent of all research objects simultaneously signs informed consent form.
Embodiment
Detection reagent is stored in advance in the inner cavity of box body 1, in use, being carried out by syringe from filler 4, rubber stopper 6
Reagent is extracted, box body 1 can be used as reaction chamber, be observed by photoscope 5, and specifically used steps are as follows:
Prepare colon cancer cases group and compares the cell of biopsy or culture
It extracts colon cancer cases group and compares the RNA of biopsy, the integrality of RNA is proved by gel electrophoresis, uses core
The concentration and purity of acid albumin instrument measurement RNA.Main operational steps are as follows:
(1) histocyte, centrifuging and taking upper cell are cracked with cellular lysate liquid BL.Detection reagent is added in upper cell,
It is lashed lysate 10 times with the disposable syringe with syringe needle.
(2) it stands after five minutes, the chloroform of 200 μ l is added, after firmly overturning centrifuge tube mixing, be stored at room temperature and be allowed to be layered,
12000g is centrifuged 5 minutes, carefully pipettes water phase into the centrifuge tube of 1.5ml.
(3) isometric isopropanol is added, after thoroughly mixing, takes out 750 μ l and moves into adsorption column, be centrifuged 30 seconds, outwell receipts
Liquid in collector moves into adsorption column in same collecting pipe, will all enter remaining in adsorption column, is centrifuged 30 seconds.It outwells
Liquid in collecting pipe moves into adsorption column in the same collecting pipe.
(4) 500 μ L RP liquid are added, are centrifuged 30 seconds.The liquid in collecting pipe is outwelled, adsorption column is moved into same collecting pipe
In.
(5) by the W3 liquid of 500 μ l, 1 minute is stood, is centrifuged 15 seconds.
(6) adsorption column is moved into a clean collecting pipe, 500 μ l W3 liquid is added, be centrifuged 15 seconds.
(7) liquid in collecting pipe is outwelled, then adsorption column is moved into same collecting pipe, is centrifuged 1 minute.
(8) adsorption column is put into the centrifuge tube of another clean 1.5ml, 50 μ l pure water is added in adsorbed film center,
After being stored at room temperature l/min, it is centrifuged 1 minute.RNA is stored in -70 DEG C.
(9) total serum IgE integrality is identified: being taken 2 μ l RNA samples at 1.5% agarose gel electrophoresis (80v, 15min), is separated
After zone, EB is dyed, and Zone electophoresis band is observed under ultraviolet lamp.
(10) with the concentration and purity of nucleic acid-protein instrument measurement RNA.
(11) building and sequencing of colon cancer cases group and control biopsy mRNA transcript profile library.
(12) transcript mRNA is purified using total serum IgE obtained above.
(13) Single-end library banking process is utilized, intestinal cancer case group and control biopsy mRNA is obtained
Transcript profile library.
(14) above-mentioned mRNA transcript profile library is subjected to high-flux sequence, to high-flux sequence sequence filter, removes low-quality
Sequence is measured, is compared using SOAP2 method with human genome.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
A variety of variations, modification, replacement can be carried out to these embodiments without departing from the principles and spirit of the present invention by understanding
And modification, the scope of the present invention is defined by the appended.
Claims (7)
1. a kind of detection kit for the early diagnosis of straight colon cancer, including box body (1) and box cover (3), it is characterised in that: institute
The upside that box cover (3) is screwed onto box body (1) is stated, the bottom of the box body (1) is provided with reflector (2), the top of the box cover (3)
End right side is provided with filler (4), round through slot is offered in the middle part of the ontology of the box cover (3), and laterally embedding in round through slot
Enter to have photoscope (5), the inner lower of the filler (4) is embedded with rubber stopper (6).
2. a kind of detection kit for the early diagnosis of straight colon cancer according to claim 1, it is characterised in that: described
Box body (1) and filler (4) integrated injection molding.
3. a kind of detection kit for the early diagnosis of straight colon cancer according to claim 1, it is characterised in that: described
The bottom of box body (1) is provided with stabilizer blade, and stabilizer blade has been embedded in rubber pad, and rubber pad is Nian Jie with stabilizer blade by bonding agent.
4. a kind of detection kit for the early diagnosis of straight colon cancer according to claim 1, it is characterised in that: described
Reflector (2) includes plastic protective plate and masking foil, and masking foil is placed in the downside of plastic protective plate, the edge of the plastic protective plate
It is Nian Jie with the intracavity bottom of box body (1).
5. a kind of detection kit for the early diagnosis of straight colon cancer according to claim 1, it is characterised in that: described
The bottom of filler (4) is provided with conical plate, and conical plate is in hollow funnel-form, and the conical plate and filler (4) are coaxial
Setting.
6. a kind of detection kit for the early diagnosis of straight colon cancer according to claim 1, it is characterised in that: described
Lateral retaining ring is provided on the downside of the inner cavity of filler (4), and lateral retaining ring is supported on the downside of rubber stopper (6).
7. a kind of application of detection kit for the early diagnosis of straight colon cancer according to claim 1, feature exist
In: storage of the detection kit for straight colon cancer early diagnosis reagent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811112982.8A CN109136080A (en) | 2018-09-25 | 2018-09-25 | A kind of detection kit and its application for the early diagnosis of straight colon cancer |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811112982.8A CN109136080A (en) | 2018-09-25 | 2018-09-25 | A kind of detection kit and its application for the early diagnosis of straight colon cancer |
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| CN109136080A true CN109136080A (en) | 2019-01-04 |
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| CN201811112982.8A Pending CN109136080A (en) | 2018-09-25 | 2018-09-25 | A kind of detection kit and its application for the early diagnosis of straight colon cancer |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111560313A (en) * | 2020-05-14 | 2020-08-21 | 保睿(上海)生物科技有限公司 | Detection kit for enterobacter microorganisms and detection method thereof |
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Application publication date: 20190104 |