CN104004067B - Antigen and related immune detection kit for detecting anti-human papilloma virus (anti-HPV) antibody - Google Patents
Antigen and related immune detection kit for detecting anti-human papilloma virus (anti-HPV) antibody Download PDFInfo
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Abstract
The present invention relates to a kind of HPV HPV oncogene proteins as antigen, the antibody for detecting anti-human papilloma virus (anti-HPV) HPV, it is selected from such as SEQ ID NO:1、SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:Amino acid sequence shown in 5 has one or more in amino acid sequences of 80% homology with them.Additionally providing includes the immunity detection reagent of above-mentioned antigen.And the application of above-mentioned antigen and kit.The antigen for detecting anti-HPV antibody of the present invention can detect the HPV antibody in blood or saliva with high sensitivity, high specific and high stability, so as to there is the oncogene-transformed Cervical intraepitheliaI neoplasia of HPV viruse and cervix cancer to bring foundation for doctor Accurate Diagnosis patient, and then take treatment in time, to prevent cancer, or cancer diffusion, mitigate patient's pain in time, the immunity detection reagent being made from it can quickly and easily detect the HPV antibody in blood or saliva exactly, suitable for large-scale promotion application.
Description
Technical field
The present invention relates to antibody test technical field, more particularly to anti-human papilloma virus (anti-HPV)（Human
Papillomavirus, HPV）Antibody test technical field, is specifically referred to a kind of for detecting anti-human papilloma virus (anti-HPV) antibody
Antigen and related immune detection kit, are used especially for the high-risk type HPV of domestic prevalence（HPV）Hypotype
HPV16 and HPV18.
Background technology
Cervix cancer is the common female malignant of second, people's papilloma virus（HPV）It is the necessary condition that it occurs.
Although crowd's HPV infection is very universal, such as：Uterine neck high cancer incidence area women infection rate still up to 20.8% after 35 years old.But more than 80%
HPV DNA positive womens will not progress to cancer or precancerous lesion.Therefore HPV DNA detections defect is can not to differentiate transient
Infect and the infection of cervical lesionses can be caused, cause higher false positive rate.Numerous studies show, HPV oncogene expression products
（Cancer protein oncoprotein）There is a very high carcinogenic activity, therefore have can be as than HPV DNA more for these biomarkers
Accurately there is specific cervical carcinoma prediction index.The expression of high-risk HPV E2, E6, E7 cancer protein (oncoprotein) and
Precancerous lesions of uterine cervix different phase is directly related.Brand-new cervical carcinoma Molecular screening index can be built using this characteristic.
Research to HPV mechanism of carcinogenesis shows, the hair of early expression albumen E6, the E7 albumen of high-risk HPV in cervical carcinoma
Played an important role in life.In the high of cervical cell pathological changes and the cervical epithelial cellses of cancer patient HPV E6 and E7 albumen it is general in
The E6 of overexpression, continuous expression and overexpression, E7 carcinogenic proteins can be with host cell modulin（Such as, P53 and Rb albumen
Deng）Interaction, inactivates tumor suppressor, causes cell immortality and vicious transformation.
Except HPV DNA detections and the detection of HPV carcinogenic proteins, blood（Body fluid）Middle HPV antibody also demonstrate that to HPV infection and
Cervical cancer pathogenesis history has very big correlation.HPV L1 blood antibody available standards ELISA method is detected, provides HPV infection letter
Breath.With the extension of HPV infection time, HPV inductions generate anti-HPV antibody, therefore can detect serum moderate resistance HPV antibody.
The male couple that Wikstrom etc. have detected the existing disease patients with condyloma acuminatum of 47 males and 32 once had a condyloma acuminatum medical history patient
The serum IgG antibody of HPV6 types and HPV11 type capsid proteins, and compared with the male of 205 agenosomia condyloma acuminatum medical histories,
As a result, the IgG antibody of serum moderate resistance HPV6 types is 35% in the male for once having condyloma acuminatum medical history, in existing disease condyloma acuminatum
For 27%, and it is 10% in control group.This shows that the appearance of anti-HPV6IgG antibody occurs in condyloma acuminatum stadium later,
The IgG positives, which also reflect, simultaneously once infected this virus.
The infection rate that Stone etc. reports using the method for 7218 HPV-16 of U.S. general population is studied, as a result
Show women positive rate 17.9%, male's positive rate 7.9%（Stone, K.M., et al., Seroprevalence of
HumanPapillomavirus Type16Infection in the United States.2002.The Journal of
Infectious Diseases2002;186:1396-402）.HPV L1 antibody ELISAs methods are coordinating HPV vaccines（Merck
Company's Gardasil preventative vaccines）Research and development and clinical test during be once widely adopted.Luevano etc. uses protein chip
The blood serum sample of example Patients with Cervical Cancer has carried out high flux detection and analysis（Luevano,M.,et al.,High-Throughput
Profling of the Humoral Immune Responses Against Thirteen Human
PapillomavirusTypes by Proteome Microarrays.2010.Virology.2010;405(1):31–40）,
The antibody profiling of 54604 kinds of albumen altogether of 13 HPV types is analyzed, the antibody of carcinogenic protein in blood is as a result indicated
There is stronger correlation with cervical carcinoma, height lesion, low lesion and asymptomatic ordinary people（Luevano,M.,et al.,
High-Throughput Profling of the Humoral Immune Responses Against Thirteen
Human Papillomavirus Types by Proteome Microarrays.2010.Virology.2010;405(1):
31–40）.
Therefore, in order to realize high sensitivity, high specific and the high stability of HPV antibody tests, it is desirable to provide new use
In the antigen of detection anti-human papilloma virus (anti-HPV) antibody.
The content of the invention
In order to solve above-mentioned problem, it is used to detect that anti-human papilloma virus (anti-HPV) resists it is an object of the invention to provide one kind
The antigen and related immune detection kit of body, this be used to detecting anti-human papilloma virus (anti-HPV) antibody antigen can with high sensitivity,
The HPV antibody in blood or saliva is detected to high specific and high stability, so as to being doctor's Accurate Diagnosis patient's uterine neck
Intraepithelial neoplasia (cin) and cervix cancer bring foundation, and then take treatment in time, to prevent cancer, or cancer diffusion, in time
Mitigate patient's pain, the immunity detection reagent being made from it can quickly and easily detect the HPV in blood or saliva exactly
Antibody, suitable for large-scale promotion application.
To achieve these goals, it is used to detect anti-human papilloma virus (anti-HPV) there is provided one kind in the first aspect of the present invention
The antigen of antibody, is characterized in, the described antigen for being used to detect anti-human papilloma virus (anti-HPV) antibody is selected from such as SEQ ID NO:1 institute
Amino acid sequence, such as SEQ ID NO shown:Amino acid sequence, such as SEQ ID NO shown in 2:Amino acid sequence shown in 3, such as
SEQ ID NO:Amino acid sequence, such as SEQ ID NO shown in 4:Amino acid sequence and SEQ ID NO shown in 5:Shown in 1
Amino acid sequence has the amino acid sequence and SEQ ID NO of 80% homology:Amino acid sequence shown in 2 is homologous with 80%
Property amino acid sequence, with SEQ ID NO:Amino acid sequence shown in 3 has the amino acid sequence and SEQ of 80% homology
ID NO:Amino acid sequence shown in 4 have 80% homology amino acid sequence and with SEQ ID NO:Amino acid shown in 5
Sequence has the one or more in the amino acid sequence of 80% homology.
It is preferred that described m+2, m+3, m+5 being used to detect the antigen of anti-human papilloma virus (anti-HPV) antibody
Position, m+6 and m+8-m+12 amino acids are Asp, Ser, Glu, Glu, Asp, Glu, Ile, Asp and Gly successively, its
Middle m >=0.
It is preferred that the described antigen for being used to detect anti-human papilloma virus (anti-HPV) antibody uses chemical synthesis or bioengineering side
Method is obtained.Example：Recombinant protein, fusion protein.
In the second aspect of the present invention there is provided a kind of immunity detection reagent, it is characterized in, including above-mentioned is used to examine
Survey the antigen of anti-human papilloma virus (anti-HPV) antibody.
It is preferred that the immunity detection reagent also includes carrier, it is described to be used to detect anti-human papilloma virus (anti-HPV) antibody
Antigen coat on the carrier.
It is preferred that the immunity detection reagent also includes the secondary antibody of the anti-human papilloma virus (anti-HPV) antibody of mark.Secondary antibody is used
To detect that the IgG antibody in human serum, IgM, or label used in IgA. can be horseradish peroxidases, alkaline phosphatase,
Fluorescence molecule FITC（Or other fluorescence labelings）, chemiluminescence detection.Detection method can be development process, fluorescent method, chemistry
Luminous, Electrochemiluminescince carries out qualitative and quantitative analysis.
More preferably, the secondary antibody of the anti-human papilloma virus (anti-HPV) antibody of described mark is the anti-human of horseradish peroxidase-labeled
The secondary antibody of papillomavirus antibody.
In the third aspect of the present invention, there is provided the above-mentioned antigen or above-mentioned for being used to detect anti-human papilloma virus (anti-HPV) antibody
Application of the immunity detection reagent in detection human body fluid in anti-human papilloma virus (anti-HPV) antibody.
It is preferred that the body fluid is blood or saliva.
The beneficial effects of the present invention are：The antigen for being used to detect anti-human papilloma virus (anti-HPV) antibody of the present invention is selected from such as
SEQ ID NO:Amino acid sequence, such as SEQ ID NO shown in 1:Amino acid sequence, such as SEQ ID NO shown in 2:Shown in 3
Amino acid sequence, such as SEQ ID NO:Amino acid sequence, such as SEQ ID NO shown in 4:Amino acid sequence and SEQ shown in 5
ID NO:Amino acid sequence shown in 1 has the amino acid sequence and SEQ ID NO of 80% homology:Amino acid sequence shown in 2
Amino acid sequence of the row with 80% homology and SEQ ID NO:Amino acid sequence shown in 3 has the amino of 80% homology
Acid sequence and SEQ ID NO:Amino acid sequence shown in 4 have 80% homology amino acid sequence and with SEQ ID NO:
Amino acid sequence shown in 5 has the one or more in the amino acid sequence of 80% homology, patient after being infected,
Can in serum and saliva of buccal cavity high specific, high sensitivity, detect the specific antibody IgM of appearance with high specificity,
IgA or IgG molecules, so as to be that doctor's Accurate Diagnosis patient Cervical intraepitheliaI neoplasia and cervix cancer bring foundation, and then
Treatment is taken in time, to prevent cancer, or cancer diffusion, mitigates patient's pain in time, is tried with the immune detection that this is made
Agent box, the HPV antibody in blood or saliva can be quickly and easily detected exactly, suitable for large-scale promotion application.
Embodiment
In order to be more clearly understood that the technology contents of the present invention, spy enumerates specific examples below detailed description.So
And, specific embodiment is merely for illustration, rather than limitation of the present invention.
First, materials and methods
1st, reagent and medicine
Horseradish peroxidase-labeled goat-anti people（Goat Anti-Human IgG, FC）, purchased from Priece Product#
31413。
Sodium chloride, potassium chloride, disodium hydrogen phosphate, potassium dihydrogen phosphate are purchased from Chemical Reagent Co., Ltd., Sinopharm Group.
Tween-20 is purchased from AMRESCO.Hyclone is purchased from the prosperous Golden Horse biotechnology development corporation, Ltd. of Beijing member.
2nd, instrument and consumptive material
Electronic balance（Plum Teller-support benefit instrument Shanghai Co., Ltd）；85-1 constant temperature blender with magnetic force（Community of Jin Tan County city
Outstanding riel Electrical Appliances Co., Ltd）；MULTISKAN MK3 ELIASAs（Thermo companies）；Ultrapure water preparation device（The U.S.
MILLPORE companies）；GZX-9240MBE digital display air dry ovens（Medical Equipment Plant of Shanghai Boxun Industrial Co., Ltd.）；
WellWASK4MK2 board-washings machine (is purchased from Thermo companies)；ELISA Plate（COSTAR companies）；Single track, multichannel pipettor（Germany
Eppendorf companies）.
3rd, main solution system
It is coated with buffer solution（PBS phosphate solutions）：Potassium dihydrogen phosphate 0.2g, 12 water disodium hydrogen phosphate 2.9g, sodium chloride
8.0g, potassium chloride 0.2g, plus distilled water, to 1L, 4 degree preserve.
Lavation buffer solution（PBS-Tween20,0.1%）：Tween-201mL, is added to 1L dilution buffers（PBS）In,
Stir and evenly mix, between tune pH value to 7.0~7.2.
Confining liquid：5%milk-PBS-Tween
Serum dilution：10%FBS-PBS-Tween
TMB shows liquid：Amresc, CAT NO.64285730, are mixed in equal volume with preceding TMB background colors colour developing A liquid and B liquid.
Zero substrate colour developing A liquid：Sodium acetate 13.6g, citric acid 1.6g, hydrogen peroxide（30%）0.3ml, distilled water is added to
500ml。
Zero substrate colour developing B liquid：Disodium ethylene diamine tetraacetate 0.2g citric acid 0.95g glycerine 50ml tetramethyl benzidines 0.2g
Distilled water adds to 500ml.
Terminate liquid（2mol/L H2SO4）：Distilled water 178.3mL, is added dropwise the concentrated sulfuric acid（98%）21.7mL.
4th, the foundation of indirect ELISA method
1）It is with PBS dilutions HPV polypeptide antigen HPV16-1/HPV16-2/HPV16-3/HPV18-1/HPV18-2 concentration
4ug/ml, coating costar lath 100ul/ hole, 4 DEG C overnight.
2）PBS washed once, and pat dry.
3）Add 5% milk powder-PBS closing, 250ul/ holes, 37 degree 3 hours.
4）PBST washed once, and pat dry.
5）10%FBS-PBST dilute serums 1：50 dilutions, oscillator is mixed.
6）The serum diluted, blank control, negative control and positive control are added, 50ul/ holes, 37 degree of incubations 1 are small
When.
7）0.1%tween-PBS is washed 5 times, is patted dry.
8）Horseradish peroxidase-labeled goat-anti people is added, is diluted with 10%FBS-PBST（1：40000）, 100ul/ holes, 37
Degree is incubated 30 minutes.
9）0.1%tween-PBS is washed 5 times, is patted dry.
10）TMB develops the color:TMBA liquid and B liquid are mixed in equal volume, 100ul/ holes, 37 degree of 20min.
11）2M H2SO4, 50ul/ holes are terminated, with ELIASA 450nm readings.
5th, kit forms：
This kit is made up of following components：
1. one piece of the costar ELISA Plates for being used to detect the antigen of anti-human papilloma virus (anti-HPV) antibody of the coating present invention
2. horseradish peroxidase-labeled goat-anti people one manages（10ul）
3.10*PBST one bottle（10ml）
One bottle of 4.TMB nitrite ion A liquid（6ml）, one bottle of B liquid（6ml）
5. one bottle of terminate liquid（6ml）
6th, envelope antigen
Polypeptide antigen is as follows
| HPV16-1 | KCDSTLRLCVQSTHVDIRTLE | SEQ ID NO：1 |
| HPV16-2 | PTLHEYMLDLQPETTDLYCYEQLNDSSEEE | SEQ ID NO：2 |
| HPV16-3 | LNDSSEEEDEIDGPAGQAEPDRAH | SEQ ID NO：3 |
| HPV18-1 | IDGVNHQHLPARRAEPQR | SEQ ID NO：4 |
| HPV18-2 | SDSEEENDEIDGVNHQHLPARRAEPQRH | SEQ ID NO：5 |
2nd, testing result
Kit selects 5 polypeptides（SEQ ID NO：1, SEQ ID NO：2, SEQ ID NO：3, SEQ ID NO：4, SEQ
ID NO：5）As coating selection, serum is detected by ELISA method.
1st, pass through ELISA method with envelope antigen and the ELISA Plate closed and detect Healthy Human Serum, the original number of detection
It is as follows according to result；
Table one：HPV blood ELISA detects normal human serum result table
2nd, 2 polypeptides are selected（SEQ ID NO：3, SEQ ID NO：5）The ELISA Plate for selecting and closing as coating leads to
ELISA method detection normal human serum is crossed, the raw data results of detection are as follows；
Table two：HPV blood ELISA detects normal human serum result table
3rd, 5 polypeptides are selected（SEQ ID NO：1, SEQ ID NO：2, SEQ ID NO：3, SEQ ID NO：4, SEQ ID
NO：5）The ELISA Plate for being coated with and closing as coating selection detects patients serum, the pathological diagnosis of patient by ELISA method
As a result as shown in table three, which part patient passes through HC2 pathological examinations, as a result as follows：
Table three：HPV blood ELISA detects patients serum's result table
4th, 3 polypeptides are selected（SEQ ID NO：1, SEQ ID NO：3, SEQ ID NO：5）As coating selection coating simultaneously
The ELISA Plate closed detects patients serum by ELISA method, and the pathological diagnosis result of patient is as shown in table four, its middle part
Patient is divided to pass through HC2 pathological examinations, it is as a result as follows：
Table four：HPV blood ELISA detects patients serum's result table
3rd, data analysis
1st, according to normal person's detected value, average value, standard variance, CUTOFF values are calculated.Respectively by 44 in table one
Reaction OD value of the normal human serum on HPVE7 polypeptides is taken the mean, and calculates standard variance SD, this kit it is determined that
The standard variance that normal person's average value adds twice is selected during CUTOFF values as the criterion of HPV yin and yang attributes, it is as a result as follows：
Cutoff calculation formula are：Cutoff=average value OD+2*SD standard variances
1.1 according to the result of table one, HPVE7（SEQ ID NO：1, SEQ ID NO：2, SEQ ID NO：3, SEQ ID NO：
4, SEQ ID NO：5）5 polypeptide detection normal human serum result statistical analysis is drawn, as a result such as following table：
Table five：HPV E7 5 polypeptide serum ELISA detection tables of data
| SEQ ID NO：1 | SEQ ID NO：2 | SEQ ID NO：3 | SEQ ID NO：4 | SEQ ID NO：5 | |
| Average value | 0.292 | 0.279 | 0.382 | 0.301 | 0.383 |
| Standard variance | 0.098 | 0.100 | 0.111 | 0.076 | 0.111 |
| cutoff | 0.487 | 0.480 | 0.604 | 0.452 | 0.605 |
1.2 according to the result of table two, HPVE7（SEQ ID NO：3 and SEQ ID NO：5）2 polypeptide detects normal human serum
As a result statistical analysis is drawn, as a result such as following table：
Table six：HPV E7 2 polypeptide serum ELISA detection tables of data
| SEQ ID NO：3 | SEQ ID NO：5 | |
| Average value | 0.393 | 0.387 |
| Standard variance | 0.104 | 0.108 |
| cutoff | 0.601 | 0.603 |
1.3 draw SEQ ID NO according to the result of table five and table six, comparative analysis：3 and SEQ ID NO：5 this 2 polypeptides
The cutoff values drawn in different experiments are consistent.
2nd, by cutoff values, the patients serum HPV of detection feminine gender, the positive is judged, what it is more than this cutoff value is
Be considered positive, as long as and a polypeptide test positive, that is, think there is HPV infection, while being contrasted with HC2 diagnostic results.
2.1 in the patients serum of detection 291 serum have HC2 diagnostic results, have 84 of case that pathological diagnosis is positive
In pathology positive patients group (clinical diagnosis " goldstandard "), Serum Antibody Detection result compares with HC2 Molecular Detection results（See
Table 7）, it is contrasted such as following table with this kit detection HPV results：（Statistics includes for positive in this testing result：
（i）Two antigen testing results have any one to be positive findings,（ii）Two antigen testing results are all positive findingses）
Table seven：According to the result of table three, for serum ELISA diagnostic results and the HC2 diagnostic result contrast tables of patient
（"+" represents that testing result is positive；"-" represents that testing result is negative）
Table eight：According to the result of table four, for serum ELISA diagnostic results and the HC2 diagnostic result contrast tables of patient
（"+" represents that testing result is positive；"-" represents that testing result is negative）
The HC2 results of the result of table seven and blood ELISA inspection results are subjected to demographics analysis, as a result such as following table：
Table nine：HC2 results and blood ELISA Detection accuracies number and ratio contrast table
Other gynecological diseases are mainly pelvic infecton, ovarian cyst, the gynecological disease such as fibroid.
The HC2 results of the result of table eight and blood ELISA inspection results are subjected to demographics analysis, as a result such as following table：
Table ten：HC2 results and blood ELISA Detection accuracies number and ratio contrast table
Other gynecological diseases are mainly pelvic infecton, ovarian cyst, the gynecological disease such as fibroid.
There are 10 serum there is no a HC2 examining reports in 2.2 detection patients serums, but pathological replacement is CIN and cervical carcinoma,
This kit detects HPV results, patient's number and positive blood tests rate comparing result such as table.
Table 11：For there is no HC2 diagnostic results, but there are the patient of pathological findings and HPV blood ELISA testing results total
Knot
（"+" represents that testing result is positive；"-" represents that testing result is negative）
Table 12：The number table of the patient blood ELISA detections of CIN without HC2 results, cervical carcinoma and cervical lesionses
| Pathology | Number（10） | SEQ ID NO：1 is positive | SEQ ID NO：3 is positive | SEQ ID NO：5 is positive |
| CINⅠ | 2 | 2 | 1 | 1 |
| CIN II, III | 7 | 4 | 5 | 5 |
| Cervical carcinoma | 1 | 0 | 1 | 1 |
2.3 by the patient blood ELISA testing results statistical number of person of all CIN I II III and cervical carcinoma and positive ratio knot
Fruit such as table 13.
Table 13：The number and accuracy rate ratio of the patient blood ELISA detections of all CIN, cervical carcinoma and cervical lesionses
Table
The aforementioned polypeptides of the present invention are the selections by inventor and examined in patient samples and normal human sample
Test card, and directly use GST-HPV16-E7 and GST-HPV118E7 total length fusion proteins（Respectively SEQ ID NO：6、SEQ
ID NO：Sequence shown in 7）Serum is detected, effect is not good, refers to following experiments data.Or
4th, experiment material and ELISA method are with above first point, simply using polypeptide（SEQ ID NO：3, SEQ ID
NO：5）While detection, also using GST-HPV16-E7 and GST-HPV118E7 total lengths fusion protein as detection antigen, with
When PBS dilutes GST-HPV16-E7, GST-HPV18-E7 total length fusion protein, concentration is 2ug/ml, and testing result is as follows：
1. according to experimental procedure above, detection part patients serum and common human serum, testing result and patient's pathology report
Accuse as shown in the table：
Five, experimental analyses
1. by using the testing result of full-length proteins and before using HPV polypeptide SEQ ID:3 and SEQ ID:5 two kinds of polypeptides
Joint detection results are contrasted, contrast such as following table：
Summarize：Summarize：HPV16-E7, HPV18-E7 total length fusion protein have certain in detection CIN during Patients with Cervical Cancer
Positive patients missing inspection, sensitivity detects poor than polypeptide.There is certain vacation when detecting common gynaecological imflammation patient and general population
Positive rate, sensitiveness detects slightly poor than polypeptide.So, HPV16-E7 and HPV18-E7 full-length proteins are for detection CIN, cervical carcinoma
Etc. being not well suited for.
There is provided high sensitivity, height based on immune biochemical basis for the blood enzyme mark immunoassay (ELISA) of the present invention
The Ag-Ab detection method of specificity and high stability.Patient can go out after being infected in serum and saliva of buccal cavity
Existing specific antibody IgM, IgA or IgG molecules.Detect special in patients serum or saliva with enzyme-labeled immunity analytic approach ELISA
The anti-HPV of property antibody molecule.The present invention passes through to HPV viruse high-risk HPV 16, the analysis of HPV18 gene orders, with biology letter
Credit analysis is ceased to antigenic theoretical prediction, and is experimentally screened, proved the meaning of clinical practice.It is multiple high by preparing
The antigen polypeptide of purity, and repeated screening, checking are carried out to preparing antigen with domestic people healthy person serum and infected person anteserum,
Obtained high sensitivity, the purpose antigen HPV16-1 of high specific and high stability, HPV16-2, HPV16-3, HPV18-1,
HPV18-2.And prove to be attached to certain surface of solid phase carriers with these polypeptide antigens, its immune response activity can be kept.Surveying
Regularly, the antigen by inspection sample and surface of solid phase carriers is made to react.The antigen that is made to be formed on solid phase carrier with the method for washing,
Antibody complex is separated with other materials, and the amount of tested substance is into one in the enzyme amount and sample that are finally incorporated on immobilization carrier
Fixed ratio.Add enzyme reaction substrate after, then detected by developing the color, thus can be made according to the depth of color reaction it is qualitative or
Quantitative analysis.The blood ELISA detection kit of the present invention is compared with DNA detections (HC2, CareHPV), and sampling is uniform, non-to invade
Property is omited, operation is more quick, simply；Expense is cheap, easily promotes, is more suitable for Chinese market；The vacation that PCR method can be avoided to bring
Positive and false negative, substantially increases accuracy and the degree of accuracy of testing result；VIA is dyed with traditional Pap smear, acetic acid,
Vaginoscopy is compared, more accurately, easily, quickly.ELISA detection method, can through the optimization in various parameters and experiment journey
It is used for the early screening of cervical carcinoma as the instrument of Large-scale Screening.
To sum up, the of the invention antigen for being used to detecting anti-human papilloma virus (anti-HPV) antibody can with high sensitivity, high specific and
The HPV antibody in blood or saliva is detected to high stability, so as to being doctor's Accurate Diagnosis patient's Cervical intraepitheliaI neoplasia
Foundation is brought with cervix cancer, and then takes treatment in time, to prevent cancer, or cancer diffusion, mitigates patient's pain in time
Hardship, the immunity detection reagent being made from it can quickly and easily detect the HPV antibody in blood or saliva exactly, suitable for big
Scale popularization and application.
In this description, the present invention is described with reference to its specific embodiment.But it is clear that can still make
Various modifications and alterations are without departing from the spirit and scope of the present invention.Therefore, specification is considered as illustrative rather than limit
Property processed.
Claims (9)
1. a kind of be used to detect the antigen of anti-human papilloma virus (anti-HPV) antibody, it is characterised in that described to be used to detect anti-human nipple
The amino acid sequence of the antigen of tumor virus antibody such as SEQ ID NO:Shown in 3.
2. a kind of be used to detect the antigen of anti-human papilloma virus (anti-HPV) antibody, it is characterised in that described to be used to detect anti-human nipple
The antigen of tumor virus antibody includes the amino acid sequence as shown in SEQ ID NO.3 and the one or more of amino being selected from the group
Acid sequence：Such as SEQ ID NO:Amino acid sequence, such as SEQ ID NO shown in 1:Amino acid sequence and such as SEQ ID shown in 4
NO:Amino acid sequence shown in 5.
3. a kind of be used to detect the antigen of anti-human papilloma virus (anti-HPV) antibody, it is characterised in that described to be used to detect anti-human nipple
The antigen of tumor virus antibody includes such as SEQ ID NO:Amino acid sequence and such as SEQ ID NO shown in 3:Amino acid shown in 5
Sequence.
4. the antigen according to claim 1 for being used to detect anti-human papilloma virus (anti-HPV) antibody, it is characterised in that described use
Obtained in the antigen of detection anti-human papilloma virus (anti-HPV) antibody using chemical synthesis or biological engineering method.
5. a kind of be used to detect the antigen of anti-human papilloma virus (anti-HPV) antibody, it is characterised in that described to be used to detect anti-human nipple
The antigen of tumor virus antibody includes such as SEQ ID NO:Amino acid sequence, such as SEQ ID NO shown in 1:Amino acid sequence shown in 3
Row, such as SEQ ID NO:Amino acid sequence and such as SEQ ID NO shown in 4:Amino acid sequence shown in 5.
6. a kind of immunity detection reagent, it is characterised in that detect anti-human including being used for according to claim 1-5 is any
The antigen of papillomavirus antibody.
7. immunity detection reagent according to claim 6, it is characterised in that the immunity detection reagent also includes carrying
Body, the described antigen coat for detecting anti-human papilloma virus (anti-HPV) antibody is on the carrier.
8. detection reagent is being prepared according to any described antigens for being used to detect anti-human papilloma virus (anti-HPV) antibody of claim 1-5
In application, the detection reagent be used for detect anti-human papilloma virus (anti-HPV) antibody in human body fluid.
9. application according to claim 8, it is characterised in that the body fluid is blood or saliva.
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| CN201310056516.3A CN104004067B (en) | 2013-02-22 | 2013-02-22 | Antigen and related immune detection kit for detecting anti-human papilloma virus (anti-HPV) antibody |
| CN201610854932.1A CN106478784B (en) | 2013-02-22 | 2013-02-22 | For detecting the antigen and related immune detection kit of anti-human papilloma virus (anti-HPV) antibody |
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| CN110596384B (en) * | 2019-10-12 | 2022-08-12 | 南京黎明生物制品有限公司 | Human papilloma virus 6 type and 11 type immunoassay kit prepared based on Cas protein and gRNA compound |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5629161A (en) * | 1991-07-13 | 1997-05-13 | Behringwerke Aktiengesellschaft | Use of HVP-16 E6 and E7-gene derived peptides to diagnose HPV-16-associated invasive cervical cancer |
| CN1433428A (en) * | 2000-04-05 | 2003-07-30 | 冲击诊断公司 | Immunological methodology for discerning human papillomavirus |
| US7026443B1 (en) * | 1999-12-10 | 2006-04-11 | Epimmune Inc. | Inducing cellular immune responses to human Papillomavirus using peptide and nucleic acid compositions |
| US7314630B2 (en) * | 2005-01-07 | 2008-01-01 | Yao-Xiong Hu | Compounds and methods of early diagnosis of cervical cancer and genital condyloma with HPV, CHSP60 tumor suppressor H-Ras, K-Ras and PTEN derived peptides modified |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SE9001705D0 (en) * | 1990-05-11 | 1990-05-11 | Medscand Ab | SET FOR DIAGNOSTICS OF VIRUS BREAKING TUMORS BY IMMUNOASSAY |
| CN1775802A (en) * | 2004-11-17 | 2006-05-24 | 宋硕 | Human papillomavirus immunogenic polypeptide, and its preparing method and use |
| CN103254307B (en) * | 2012-02-16 | 2015-03-04 | 艾托金生物医药（苏州）有限公司 | HPV18E7 monoclonal antibody, hybridoma cell strain and application |
-
2013
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-
2014
- 2014-02-21 WO PCT/CN2014/072407 patent/WO2014127741A1/en active Application Filing
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5629161A (en) * | 1991-07-13 | 1997-05-13 | Behringwerke Aktiengesellschaft | Use of HVP-16 E6 and E7-gene derived peptides to diagnose HPV-16-associated invasive cervical cancer |
| US7026443B1 (en) * | 1999-12-10 | 2006-04-11 | Epimmune Inc. | Inducing cellular immune responses to human Papillomavirus using peptide and nucleic acid compositions |
| CN1433428A (en) * | 2000-04-05 | 2003-07-30 | 冲击诊断公司 | Immunological methodology for discerning human papillomavirus |
| US7314630B2 (en) * | 2005-01-07 | 2008-01-01 | Yao-Xiong Hu | Compounds and methods of early diagnosis of cervical cancer and genital condyloma with HPV, CHSP60 tumor suppressor H-Ras, K-Ras and PTEN derived peptides modified |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106478784B (en) | 2019-10-25 |
| WO2014127741A1 (en) | 2014-08-28 |
| CN106478784A (en) | 2017-03-08 |
| CN104004067A (en) | 2014-08-27 |
| CN106632617B (en) | 2020-07-03 |
| CN106632617A (en) | 2017-05-10 |
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