CN109856401A - A kind of E7 protein detection kit and its detection method and application - Google Patents
A kind of E7 protein detection kit and its detection method and application Download PDFInfo
- Publication number
- CN109856401A CN109856401A CN201910169302.4A CN201910169302A CN109856401A CN 109856401 A CN109856401 A CN 109856401A CN 201910169302 A CN201910169302 A CN 201910169302A CN 109856401 A CN109856401 A CN 109856401A
- Authority
- CN
- China
- Prior art keywords
- detection
- protein
- application
- kit
- detection kit
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of E7 protein detection kit and its detection method and application, E7 protein detection kit, including following component: magnetic particle, anti-reagent, protein delivery agent, E7 calibration object, E7 quality-control product, cleaning solution and substrate solution is immunized in E7.The advantages that kit and detection method of the invention has stability good, high sensitivity, reproducible, the detection architecture being capable of random loading, any combination, it is continuous to detect, while substantially reducing detection time, it can complete within 1 hour 60-70 test, and it is simple to operate, it is truly realized detection full-automation, avoids manual operation bring result error, cervical carcinoma testing cost is reduced, large-scale promotion application is suitable for.
Description
Technical field
The present invention relates to protein detection technology field, more particularly to a kind of E7 protein detection kit and its inspection
Survey methods and applications.
Background technique
Cervical carcinoma is the second common cancer diagnosis in women, and in the case where 99.7% with high-risk human papilloma
Virus infection is related, and there are about 500000 cervical carcinomas in the whole world every year, and nearly 200000 people dies of cervical carcinoma.HPV has more than 100 kinds
Different separation strains, according to them and cervical carcinoma or with benign cervical lesions or the relevance of atypical hyperplasia by broadly again
It is divided into high-risk and low danger hypotype.High-risk type HPV includes HPV16,18,31,33,35,39,45,51,52,56,58,59,68 etc.
Hypotype, especially HPV16 and 18 types related to the generation of cervical carcinoma and epithelium of cervix uteri inner height lesion (CIN II/III),
Middle HPV16 accounts for 50% or more.
Human papilloma virus (H μm of an Papillomavir μ s, HPV) is a kind of thermophilic epithelial virus, shares 3 genes
District's groups are at including early stage area (area Early Region, E), late region (area Late Region, L) and noncoding region
(Uncoding Region, UCR) or upstream regulatory region (URR).The area E is E6, E7, E1, E2, E3, E4 and E5 totally 7 in order
Gene participates in the duplication, transcription, the gene for encoding virus protein, the high copy number for maintaining intracellular virus of viral DNA, wherein
E6 and E7 is the Analyses of major carcinogens in mainstream gene of HPV, related with virocyte transformation function and carcinogenicity.E6 and E7 albumen makes tumor suppressor
Proteins p53 and pRB passivation release cell cycle control respectively and inhibit Apoptosis.Therefore, for measuring the HPV in tumour
The best approach of state is the E6/E7 albumen measured in tumour cell.
The method that predominantly detects of human papilloma virus (HPV) includes in situ hybridization at present, the direct trapping of DNA and all kinds of
PCR method.Real-time fluorescence quantitative PCR (qPCR, the Real- that the mid-90 in last century grows up on the basis of traditional PCR
Time Q μ antitative PCR) technology realizes leap of the PCR from qualitative to quantitative, with its high specificity, high sensitivity,
The advantages that reproducible, quantitative accurate, speed is fast, totally-enclosed reaction, becomes the important tool in molecular diagnosis research.But
The above method only detects whether to have infected human papilloma virus (HPV) at the genetic level, as the table of related carcinogenic protein
Up to whether, Real-Time Fluorescent Quantitative PCR Technique can not provide accurate judgement, and this method needs necessary instrument real-time fluorescence
Quantitative PCR apparatus, expensive, testing cost is high.
Another cervical carcinoma detection method is cervical cytology (TCT)；Cytology detects (TCT) result and generates ASCUS people
Group's (Atypical Cells ofUndetermined Significance, atypical squamous cell are unable to meaning), no
Precancerous lesion or diagnosis of cervical cancer foundation can be provided, cervical disease or HPV infection are only detected.Therefore, it is desired nonetheless to E7 is detected, it is right
CIN 1-3 precancerous lesion is assessed.Cytolgical examination (TCT) is often due to inspection personnel's level uneven and technology feature itself
Etc. factors, cause false positive rate higher.
Therefore it provides a kind of E7 protein detection kit and its detection method, are those skilled in the art's urgent need to resolve
Problem.
Summary of the invention
In view of this, sensibility is high the present invention provides a kind of E7 protein detection kit and its detection method, specificity
By force, accuracy is high.
To achieve the goals above, the present invention adopts the following technical scheme:
A kind of E7 protein detection kit, including following component: magnetic particle, anti-reagent, protein delivery agent, the school E7 is immunized in E7
Quasi- product, E7 quality-control product, cleaning solution and substrate solution.
Further, a kind of detection method of E7 protein detection kit, the specific steps are as follows:
(1) women cervical exfoliated cell is acquired, is placed in centrifuge tube；The women did not had sexual life and vagina within three days
Medication；
(2) the protein delivery agent of 1.0ml is added, be vortexed concussion 1-2 minutes；
(3) it takes out and is placed in 1.5ml centrifuge tube, be put into -20 refrigerator freezing 6-8 minutes；
(4) it is redissolved 5 minutes in 37 degree of incubators, 4000 revs/min, 4 degree are centrifuged 5 minutes, the supernatant after taking centrifugation, as
Sample is detected；
(5) it is poured into sample cell after redissolving E7 calibration object, is put into instrument in order；
(6) the anti-reagent of 30 μ l E7 calibration objects/E7 quality-control product/sample and 60 μ l is added in every hole, and 37 DEG C of incubation 30min are used
Exhaust supernatant after cleaning solution cleaning 4 times, is added after magnetic particle is immunized in 30 μ l E7 and places 5 minutes, after cleaning 4 times with cleaning solution
The supernatant that exhausts reads RLU after 1 minute after 200 μ l substrate solutions are added；
(7) using calibration object concentration as abscissa, RLU value is ordinate, obtains standard song using 4-parameter logistic fit
Line；
(8) E7 protein content in sample to be examined is calculated according to standard curve.
Further, a kind of application of E7 protein detection kit, the kit is used to detect the TCT positive and DNA is high-risk
The high-risk positive crowd shunted of the crowd or TCT feminine gender and DNA that the positive shunts；Lesion is detected by detection E7 albumen, including
Following steps: cervical exfoliated cell, b a) are detected) if a) being the positive, indicate that women carries out gynecatoptron detection；If c) a)
For feminine gender, then indicate that women often periodically carries out cytology detection or detected with the present invention.
Further, a kind of application of E7 protein detection kit, the kit is for detecting TCT Positive Populations；Pass through
Detection E7 albumen detects lesion, comprising the following steps: a) detects cervical exfoliated cell, b) if a) being the positive, indicate female
Property carry out gynecatoptron detection；If c) a) being feminine gender, indicate that women often periodically carries out cytology detection or examined with the present invention
It surveys.
Further, a kind of application of E7 protein detection kit, the kit are used for crowd after detection technique, postoperative crowd
Including postoperative or recurrence crowd；Lesion is detected by detection E7 albumen, comprising the following steps: a) detection uterine neck is de- before surgery
Fall cell, record testing result, b) periodic detection cervical exfoliated cell after surgery, record testing result, c) if b) than a)
Testing result increase, then indicate the postoperative failure of women or palindromia, d) if b) than a) testing result reduce, indicate
Women successful surgery is needed often periodically to carry out cytology detection or be detected with the present invention.
Further, a kind of application of E7 protein detection kit, the kit is for detecting TCT and DNA Population with Negative；
Lesion is detected by detection E7 albumen, comprising the following steps: a) detects cervical exfoliated cell, b) if a) being the positive, refer to
Show that women carries out gynecatoptron detection；If c) a) being feminine gender, indicate that women often periodically carries out cytology detection or sent out with this
Bright detection.
Further, a kind of application of E7 protein detection kit, the kit for detect HPV16,18,31,33,
35,39,45,51,52,56,58,59 or 68 high-risk type HPV.
Further, a kind of application of E7 protein detection kit, by sample to be detected through the E7 protein assay reagent
Box is detected, and is carried out four parameter fittings with concentration value with luminous value and is analyzed testing result, and then determination is to be detected
The gradient of infection of sample.
The meaning that the present invention is detected using E7 protein specific antibody:
(1) when HPV genomic instability, E7 is mixed into genome (integration), and E2 can be destroyed, and function is lost；E7
When destroying cell, it will usually which canceration can just be caused, this is the root for leading to cervical carcinoma lesion when E7 is largely movable by generating E7 albumen
This reason；
(2) E7 is the oncogene of HPV viruse, generates E7 albumen by great expression, causes cell to occur high-level
Precancerous lesion is until canceration；The correct processing of precancerous lesion is the key that cervical carcinoma detection quality assurance, because cervical carcinoma detects
Free-revving engine be not only to find out cervical carcinoma, and be timely discovery precancerous lesion and it blocked to progress to cervical carcinoma；It is not at the same level
All there are three types of development trends for other precancerous lesion: lapsing to, continues, is in progress, be appropriately determined the development trend of precancerous lesion, is high-quality
The key for measuring screening treatment, not only can avoid unnecessary over-treatment, but also prevent from failing to pinpoint a disease in diagnosis；
(3) target spot detection being carried out to cervical carcinoma using E7 albumen, directly determines and precancerous lesion whether occurs, sensibility is high,
High specificity, directly reflection cervical lesions trend；
(4) the cervical carcinoma detection based on antibody, substantially reduces false positive rate, improves diagnosis accuracy, facilitate subsequent control
It treats；
(5) E7 albumen inhibits normal apoptotic process, and inducing cell division cycle is accelerated, and papilloma change occurs；Detect E7
Albumen provides specific judgement for the cancer cell variation of cervical carcinoma；
(6) E7 protein overexpression provides theoretical foundation for cervical carcinoma lesion and CIN prediction；
(7) use scope is wide, is suitble to early shunting detection, auxiliary diagnosis and the Follow-up After of cervical carcinoma；
(8) E7 detection effectively avoids in DNA cloning as caused by pollution without detecting target purification, reverse transcription, amplification
False positive issue；
(9) it can be used for detecting that a variety of HPV high jeopardize the E7 albumen of middle danger hypotype；
(10) a variety of HPV high can be detected simultaneously and jeopardize middle danger hypotype, be not necessarily to repeated detection, can reduce required sample size and can
Effectively improve detection efficiency；
(11) Automated Chemiluminescence system can increase clinical precision and simplicity, reduce human error；
(12) detection speed is fast, and the time of the kit detection E7 albumen is 20-30min.
It can be seen via above technical scheme that compared with prior art, the present disclosure provides a kind of E7 Protein Detections
Kit and its detection method and application, have the advantages that
(1) the advantages that kit of the invention and detection method have stability good, high sensitivity, reproducible, the inspection
Survey system can random loading, any combination is continuous to detect, while substantially reducing detection time, can complete 60-70 within 1 hour
A test, and it is simple to operate, it is truly realized detection full-automation, avoids manual operation bring result error,
Cervical carcinoma testing cost is reduced, large-scale promotion application is suitable for.
(2) testing result of the invention is compared with the traditional method, and accordance is consistent, and the sensitivity of E7 protein positive is
89.3%, specificity 59.1% reduces DNA false positive rate 35%.
(3) E7 protein detection kit of the present invention and detection method can be used for detecting that a variety of HPV high jeopardize middle danger hypotype
E7 albumen, directly detect cervical carcinoma precancerous lesion, avoid directly detecting false positive caused by HPV viruse；It can detect simultaneously
A variety of HPV high jeopardize middle danger hypotype, are not necessarily to repeated detection, can reduce required sample size and can effectively improve detection efficiency；Entirely certainly
Dynamic chemiluminescence detection system can increase clinical precision and simplicity, reduce human error；
(4) detection method of E7 albumen of the present invention is suitable in outlying district, not the gynecological tumor doctor of special training
Under conditions of inspection, it is easy to operate, testing result, interpretation result are detected and directly displayed by Full-automatic chemiluminescence apparatus.
(5) detection method of E7 albumen of the present invention can be accurate when in conjunction with traditional Pap smear/detection of nucleic acids
Testing result is obtained, false positive and false negative are avoided.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
The embodiment of invention for those of ordinary skill in the art without creative efforts, can also basis
The attached drawing of offer obtains other attached drawings.
Fig. 1 attached drawing is the present invention using calibration object concentration as abscissa, and luminous value is ordinate, using four parameter logistic fits
The standard curve that method obtains.
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.
Embodiment 1
1, the preparation of magnetic particle is immunized in E7
1) magnetic particle buffer is immunized, for preparing 1L:
(1) according to dose volume amount, prepare clean container tool, the purified water of 700ml is added；
(2) Tris 5.6g, NaCl 7.5g, NaN are weighed30.5g is mixed, is mixed in room temperature on magnetic suspension device in container
1 hour；
(3) detection pH value of solution is in 9-10 or so；
(4) adjustment pH value is 7.5；
(5) TW8020ml, very much electronic balance precise BSA 1g are measured with pipettor, it is mixed in room temperature on magnetic suspension device
Even 1 hour；
(6) constant volume is to 1L.
2) preparation method of magnetic particle is immunized in E7
(1) taking 1 μm of diameter, volume is 0.1ml carboxyl magnetic bead in the clean bottle of 3ml, with 1ml without amine coupling buffer repeatedly
Cleaning 5 times, and separated with magnetic patch, it inhales and abandons supernatant, adjustment final volume is 1.0ml；
(2) plus mouse anti-FITC 0.4ml it, in mass ratio, respectively plus each 5mg of EDC and NHS, mixes well, it is small that room temperature mixes 4
When；
(3) after washing 5 times with 0.01M PBS 5ml, adjustment final volume is 1ml, dilute with immune 5-10 times of magnetic particle buffer
It releases.
2, the preparation of anti-reagent
1) preparation of anti-reagent buffer, for preparing 1L
(1) according to dose volume amount, prepare clean container tool, the purified water of 700ml is added；
(2) Tris 5.6g, NaCl 7.5g, NaN are weighed30.5g is mixed, is mixed in room temperature on magnetic suspension device in container
1 hour；
(3) pH value of solution is detected, in 9-10 or so；
(4) adjustment pH value is 7.0-7.5；
(5) TW8010ml, very much electronic balance precise BSA 25g are measured with pipettor, in room temperature on magnetic suspension device
It mixes 1 hour；
(6) constant volume is to 1L.
2) method of alkaline phosphatase (AP) label E7 antibody
(1) alkaline phosphatase of 1mg is dissolved in the SMCC solution of 10mol/L, after mixing, is stored at room temperature reaction 2 hours；
(2) desalination is carried out on AKTA protein purification instrument with G25 chromatographic column, and collects sample；
(3) it takes 2mg E7 antibody-solutions to be dissolved in the DTT solution of 1mol/L, reaction 1 hour is stored at room temperature after mixing；
(4) desalination is carried out on AKTA protein purification instrument with G25 chromatographic column, and collects sample；
(5) by the sample of collection, that is, the AP activated is mixed with antibody according to 1:1, and after mixing, 2-8 DEG C of standing reacts 18h；
(6) 10 μ l 2 mercapto ethanols are taken, be added in 5ml ultrapure water and are mixed, are added in coupling reaction liquid, rear chamber is mixed
Temperature stands reaction 1 hour；
(7) product is isolated and purified on AKTA protein purification instrument with 200 chromatographic column of S μ perdex, collects a peak
With two peaks；
(8) production concentration is measured with ultraviolet specrophotometer, and is saved with anti-reagent buffer in 4 degree；With anti-when use
Reagent buffer is configured to the working concentration of 1 μ g/ml.
3) method of fluorescein isothiocynate (FITC) label E7 antibody
(1) first by E7 antibody with anti-reagent buffer displacement 3-4 times；
(2) in E7 antibody: FITC=1:10 ratio, precise FITC, and it is molten that FITC solution is slowly added into antibody
In liquid, it is protected from light room temperature connection reaction 18h on rotary mixer, obtains connection product；
(3) by connection product through Sephadex G25 column chromatographic purifying desalination, eluent is the PBS of pH7.4, collects and produces
Object；
(4) production concentration is measured with ultraviolet specrophotometer, and with anti-reagent buffer in 4 degree of preservation；With anti-when use
Reagent buffer is configured to the working concentration of 1.1 μ g/ml.
4) preparation of anti-reagent
It 2) will be mixed with 3) the middle product collected according to 1:1~1:2.
3, protein delivery agent
1. 0.85% NaCl, 2. surfactant Tween 80 or NP400.05-1.5%, 3. protease includes being dissolved in
The 0.06-2 μ g/ml Aprotinin of PBS and or 0.5-2 μ g/ml leupetin.
4, the preparation of E7 calibration object
Calibration object preparing process is that the HPV-16 E7 of 95ng/ μ g is added in Matrix buffer according to conventional preparation method,
And be freeze-dried according to the cold dry process conditions of E7, it is redissolved with the purified water of 0.5ml.
Calibration object Matrix buffer is 0.01-0.05M phosphate buffer；0.5-1M TRIS buffer is added 1%
1% BSA is added in BSA or carbonate buffer solution.
5, the preparation of E7 quality-control product
The HPV-16 E7 of 95ng/ μ g is added in Matrix buffer, and is freeze-dried according to the cold dry process conditions of E7,
It is redissolved with the purified water of 1.0ml.
6, the preparation of cleaning solution
(1) according to dose volume amount, prepare clean container tool, the purified water of 700ml is added；
(2) Tris 5.6g, NaCl 7.5g, NaN are weighed30.5g is mixed, is mixed in room temperature on magnetic suspension device in container
1 hour；
(3) pH value of solution is detected, in 9-10 or so；
(4) adjustment pH value is 7.0-7.5；
(5) TW8020ml is measured with pipettor, is mixed 1 hour in room temperature on magnetic suspension device；
(6) constant volume is to 1L.
7, the preparation of substrate solution
(1) according to dose volume amount, prepare clean container tool, the purified water of 700ml is added；
(2) Tris 12.11g, NaN are weighed30.5g is mixed in container, is mixed 1 hour in room temperature on magnetic suspension device；
(3) solution PH is detected, 10 or so；
(4) adjustment pH value is 7.0-7.5；
(5) TW8020ml, acridinium ester 2g are measured with pipettor, is mixed 1 hour in room temperature on magnetic suspension device；
(6) constant volume is to 1L.
The detection of 2 cervical samples of embodiment
1) women cervical samples (patient did not had sexual life and vagina medicinal within three days) is taken:
Doctor see and treat patients with specula opening vagina, brush is placed on patient's cervix opening zone of transformation, turns five clockwise by exposure uterine neck
The brush head careful of sample brush is put into empty centrifuge tube by circle, is avoided the brush head of sampling from rubbing outside bottle and is located, closes the lid
Son.
2) sample process
Brush head, is carefully placed into 50ml centrifuge tube by this sample collection human body cervical exfoliated cell after sampling；1.0ml is added
Protein delivery agent, be vortexed concussion 1-2 minute, take out be placed in 1.5ml centrifuge tube, be put into -20 refrigerator freezing 6-8 minutes, in
37 degree of incubators redissolve 5 minutes, and 4000 revs/min, 4 degree are centrifuged 5 minutes, to be detected after the supernatant after centrifugation is mixed.
3) prepare before experiment
(1) reagent is taken out to balance at room temperature 30 minutes；
(2) full-automatic illumination instrument is opened, substrate solution and cleaning solution, normal operation are packed into.
4) experimental method
By full-automatic illumination instrument specification requirement, anti-reagent and immune magnetic particle are sequentially placed into specified position, and will
Calibration object pours into sample cell after redissolving, and is put into instrument in order；30 μ l calibration objects/quality-control product/sample and 60 μ l are added in every hole
Anti- reagent, 37 DEG C of incubation 30min exhaust supernatant after cleaning 4 times, are added after magnetic particle is immunized in 30 μ l and place 5 minutes, cleaning
After 200 μ l substrate solutions are added in the supernatant that exhausts after 4 times, RLU is read after 1 minute.
Using calibration object concentration as abscissa X-axis, calibration object luminous intensity RLU value is ordinate, using four parameter fitting methods
Standard curve is established, as a result as shown in Figure 1, calibration curve equation is as follows:
Y=314363.1584+ (34852785.0713-314363.1584)/(1+ (x/388.3734) ^-1.2737)
Coefficient R=0.997884.
E7 protein content in sample to be examined is calculated according to standard curve.
5) kit results interpretation
It detects normal person's cervical exfoliated cell and average value, standard variance, Jin Erji is calculated according to normal person's detected value
Calculate Cutoff value:
Cutoff calculation formula are as follows: Cutoff=average value+2*SD standard variance；
As a result as follows:
When E7 protein content is less than 4.21ng/ml in cervical exfoliated cell sample, testing result is feminine gender；It is greater than
When 4.21ng/ml, testing result is the positive.
The present invention detects patient's cervical exfoliated cell and amounts to 206, and it is 184 that kit E7 Protein Detection result, which is the positive,
Example, diagnostic sensitivity are up to 89.3%.
The foregoing description of the disclosed embodiments enables those skilled in the art to implement or use the present invention.
Various modifications to these embodiments will be readily apparent to those skilled in the art, as defined herein
General Principle can be realized in other embodiments without departing from the spirit or scope of the present invention.Therefore, of the invention
It is not intended to be limited to the embodiments shown herein, and is to fit to and the principles and novel features disclosed herein phase one
The widest scope of cause.
Claims (8)
1. a kind of E7 protein detection kit, which is characterized in that including following component: magnetic particle, anti-reagent is immunized in E7, and albumen is released
Put agent, E7 calibration object, E7 quality-control product, cleaning solution and substrate solution.
2. a kind of detection method of E7 protein detection kit according to claim 1, which is characterized in that specific steps are such as
Under:
(1) women cervical exfoliated cell is acquired, is placed in centrifuge tube；
(2) the protein delivery agent of 1.0ml is added, be vortexed concussion 1-2 minutes；
(3) it takes out and is placed in 1.5ml centrifuge tube, be put into -20 refrigerator freezing 6-8 minutes；
(4) it is redissolved 5 minutes in 37 degree of incubators, 4000 revs/min, 4 degree are centrifuged 5 minutes, the supernatant after taking centrifugation, as sample
It is detected；
(5) it is poured into sample cell after redissolving E7 calibration object, is put into instrument in order；
(6) the anti-reagent of 30 μ l E7 calibration objects/E7 quality-control product/sample and 60 μ l, 37 DEG C of incubation 30min, with cleaning are added in every hole
Exhaust supernatant after liquid cleaning 4 times, is added after magnetic particle is immunized in 30 μ l E7 and places 5 minutes, exhausts after cleaning 4 times with cleaning solution
Supernatant reads RLU after 1 minute after 200 μ l substrate solutions are added；
(7) using calibration object concentration as abscissa, RLU value is ordinate, obtains standard curve using 4-parameter logistic fit；
(8) E7 protein content in sample to be examined is calculated according to standard curve.
3. a kind of application of E7 protein detection kit according to claim 1, which is characterized in that the kit is used for
Detect the TCT positive and the high-risk positive crowd shunted of DNA or TCT feminine gender and the high-risk positive crowd shunted of DNA.
4. a kind of application of E7 protein detection kit according to claim 1, which is characterized in that the kit is used for
Detect TCT Positive Populations.
5. a kind of application of E7 protein detection kit according to claim 1, which is characterized in that the kit is used for
Crowd after detection technique.
6. a kind of application of E7 protein detection kit according to claim 5, which is characterized in that postoperative crowd's packet
Include postoperative or recurrence crowd.
7. a kind of application of E7 protein detection kit according to claim 1, which is characterized in that the kit is used for
Detect TCT and DNA Population with Negative.
8. a kind of application of E7 protein detection kit according to claim 1, which is characterized in that the kit is used for
Detect the high-risk type of HPV16,18,31,33,35,39,45,51,52,56,58,59 or 68 HPV.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910169302.4A CN109856401A (en) | 2019-03-06 | 2019-03-06 | A kind of E7 protein detection kit and its detection method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910169302.4A CN109856401A (en) | 2019-03-06 | 2019-03-06 | A kind of E7 protein detection kit and its detection method and application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109856401A true CN109856401A (en) | 2019-06-07 |
Family
ID=66899977
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910169302.4A Pending CN109856401A (en) | 2019-03-06 | 2019-03-06 | A kind of E7 protein detection kit and its detection method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109856401A (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1928560A (en) * | 2006-05-26 | 2007-03-14 | 深圳市新产业生物医学工程有限公司 | Immune analysis reagent for magnetic separation alkali phosphatase enzyme mark and analytical detection method for same |
| CN102498217A (en) * | 2009-04-20 | 2012-06-13 | 阿波维塔公司 | Antibodies specific to E6 proteins of HPV and use thereof |
| CN106610378A (en) * | 2016-02-26 | 2017-05-03 | 弗雷米德生物医药技术（天津）有限公司 | Detection kit for HPV16-E7 protein |
| US20170315124A1 (en) * | 2014-09-26 | 2017-11-02 | Attogen Biomedical (Suzhou) Inc. Ltd. | Cervical cancer-related hpv e7 protein monoclonal antibody and use thereof |
| WO2018102982A1 (en) * | 2016-12-06 | 2018-06-14 | 亳州市新健康科技有限公司 | Poct fluorescence quantitative detection kit for hpv16 e7 protein and application thereof |
| CN108226515A (en) * | 2017-11-28 | 2018-06-29 | 泰州泽成生物技术有限公司 | A kind of kit and its test method for measuring sex hormone binding globulin content |
-
2019
- 2019-03-06 CN CN201910169302.4A patent/CN109856401A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1928560A (en) * | 2006-05-26 | 2007-03-14 | 深圳市新产业生物医学工程有限公司 | Immune analysis reagent for magnetic separation alkali phosphatase enzyme mark and analytical detection method for same |
| CN102498217A (en) * | 2009-04-20 | 2012-06-13 | 阿波维塔公司 | Antibodies specific to E6 proteins of HPV and use thereof |
| US20170315124A1 (en) * | 2014-09-26 | 2017-11-02 | Attogen Biomedical (Suzhou) Inc. Ltd. | Cervical cancer-related hpv e7 protein monoclonal antibody and use thereof |
| CN106610378A (en) * | 2016-02-26 | 2017-05-03 | 弗雷米德生物医药技术（天津）有限公司 | Detection kit for HPV16-E7 protein |
| WO2018102982A1 (en) * | 2016-12-06 | 2018-06-14 | 亳州市新健康科技有限公司 | Poct fluorescence quantitative detection kit for hpv16 e7 protein and application thereof |
| CN108226515A (en) * | 2017-11-28 | 2018-06-29 | 泰州泽成生物技术有限公司 | A kind of kit and its test method for measuring sex hormone binding globulin content |
Non-Patent Citations (1)
| Title |
|---|
| 张赟等主编: "磁分离酶免疫技术", 《细胞和分子免疫学实用实验技术》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Viscidi et al. | Serologic response in human papillomavirus‐associated invasive cervical cancer | |
| Kurman et al. | Immunoperoxidase localization of papillomavirus antigens in cervical dysplasia and vulvar condylomas | |
| Kirgan et al. | Association of human papillomavirus and colon neoplasms | |
| Gravitt et al. | New technologies in cervical cancer screening | |
| Barcelos et al. | Atypical squamous cells of undetermined significance: Bethesda classification and association with Human Papillomavirus | |
| US5968522A (en) | Immunogenic compositions and antibodies directed against human papillomavirus type 49 (HPV 49), type 50 (HPV 50), type 54 (HPV 54), and type 55 (HPV 55). | |
| CN105087827B (en) | Detection primer, probe and the kit of 16 kinds of type HPV viruses | |
| JP2010136729A (en) | Assessment of human papilloma virus-related disease | |
| Lonky et al. | Triage of atypical squamous cells of undetermined significance with hybrid capture II: colposcopy and histologic human papillomavirus correlation | |
| Sonnex | Human papillomavirus infection with particular reference to genital disease. | |
| CN107177699A (en) | A kind of human papilloma virus（HPV）Parting quick determination method | |
| CN104818342B (en) | Detection kit, detection architecture and method for 19 kinds of high-risk human mammilla papillomavirus (HPV) | |
| CN101186636A (en) | Human papillomavirus shell protein L1 short peptide and application thereof | |
| Arezzo et al. | HPV-negative cervical cancer: a narrative review | |
| WO2012088195A2 (en) | Device and methods for the detection of cervical disease | |
| Xiao et al. | Observations on the expression of human papillomavirus major capsid protein in HeLa cells | |
| CN104017906B (en) | A kind of HPV high-risk-type parting fluorescence PCR detection kit | |
| CN112575123B (en) | Primer combination, probe combination and human papillomavirus nucleic acid detection kit | |
| Pagliusi et al. | International standard reagents for HPV detection | |
| Wang et al. | Updates in cervical cancer screening guidelines, the Bethesda system for reporting cervical cytology, and clinical management recommendations | |
| Komolmalai et al. | Human papillomavirus 16 and 18 infection in oral cancer in Thailand: a multicenter study | |
| CN109856401A (en) | A kind of E7 protein detection kit and its detection method and application | |
| KR100671825B1 (en) | Kit for diagnosis of cervical cancer by human papilloma virus | |
| Sui et al. | Gain of the human telomerase RNA gene TERC at 3q26 is strongly associated with cervical intraepithelial neoplasia and carcinoma | |
| McDougall et al. | Methods for diagnosing papillomavirus infection |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20190902 Address after: 300 000 Huayuan Industrial Zone, Tianjin Binhai High-tech Zone Applicant after: Jinchao Miao Address before: Room 201, Building 23, Phoenix, Meteorological Station Road, Hexi District, Tianjin Applicant before: Miao Jinchao |
|
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190607 |