CN104818342B - Detection kit, detection architecture and method for 19 kinds of high-risk human mammilla papillomavirus (HPV) - Google Patents
Detection kit, detection architecture and method for 19 kinds of high-risk human mammilla papillomavirus (HPV) Download PDFInfo
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Abstract
The invention discloses a kind of detection kit, detection architecture and method of detection HPV (HPV) 19 kinds of high-risk-types, wherein 19 kinds of high-risk HPV s include HPV16,18,26,31,33,35,39,45,51,52,53,56,58,59,66,68,70,73,82；The present invention is expanded using a plurality of universal primer to purpose fragment, amplified fragments are detected in real time using special fluorescence probe, using universal primer, the complexity of system primer consumption and system are reduced, cost is saved, 10 primer collective effects detect all high-risk-types；20 specific probe makes a distinction to each type.The present invention can detect 19 kinds of high-risk HPVs in same reaction tube, and sensitivity is high, and high specificity is simple and efficient to handle, meanwhile, for patients, save examination expense.
Description
Technical field
The invention belongs to biological technical field, it is used to detecting 19 kinds of high-risk human mammilla papillomavirus more particularly to a kind of
Primer, probe, kit and its method.
Background technology
HPV (HPV) is a kind of papilloma vacuolating virus A category for belonging to papovaviridae, is spherical DNA
Virus, can cause the scaly epithelium of human body skin mucous membrane to be bred, and the mankind are its unique hosts.Have determined HPV types at present
190 kinds are had been over, about more than 50 plant the stratified squamous epithelium of orientation infection human body genital tract skin and mucous membrane.Clinically,
HPV is divided to for high-risk-type and the major class of low risk two according to HPV different subtype pathogenicities big I.High-risk-type is except can cause outer reproduction
Outside device wart, it is often more important that cause external genital organs cancer, cervical carcinoma and high-grade cervical intraepithelial neoplasia (cin), high-risk-type is to cause uterine neck
The main arch-criminal of cancer, about 99.7% cervical carcinoma is caused by the lasting or repeated infection of high-risk human mammilla papillomavirus；And it is low
It is low in epithelium of cervix uteri that danger type HPV (HPV) mainly causes the benign lesions such as the condyloma acuminatum of men and women's external genital organs to include
Lesion (CIN I) is spent, if benign lesion is diagnosed and treated not in time also the possibility for developing into canceration.Therefore, targetedly
Complete detection is carried out to high-risk HPV, had great importance to the early diagnosis and therapy of cervical carcinoma.
Traditional cytology detection method is only capable of detecting the change in terms of cytology, its sensitivity is low, poor specificity, vacation
Negative rate and false positive rate are high, and can not carry out parting to HPV.
The content of the invention
It is an object of the invention to provide primer, the probe for detecting 19 kinds of high-risk HPV s, 19 kinds of high-risk HPV s
Including HPV16,18,26,31,33,35,39,45,51,52,53,56,58,59,66,68,70,73,82；Its primer sequence is such as
Shown in table 1, as shown in table 2, interior label primer probe is as shown in table 3 for probe sequence.
The primer of table 1 and sequence
The probe of table 2 and sequence
| HPV types | Probe title | Probe sequence | Sequence number |
| HPV16 | HPV16-P1-C | CGCAGTACAAATATGTCATTATGTGCTGCC | SEQ ID NO:11 |
| HPV18 | HPV18-P3-C | TTAACAATATGTGCTTCTACACAGTCTCCTGTACC | SEQ ID NO:12 |
| HPV26 | HPV26-P1-F | GTTGATACCACCCGCAGTACTAACCTTACC | SEQ ID NO:13 |
| HPV31 | HPV31-P1-F | CCAATATGTCTGTGTGTGCTGCAATTGC | SEQ ID NO:14 |
| HPV33 | HPV33-P1 | CCACTCGCAGTACTAATATGACTTTATGCACAC | SEQ ID NO:15 |
| HPV35 | HPV35-P1-F | GTCTGTGTGTTCTGCTGTGTCTTCTAGTGACAG | SEQ ID NO:16 |
| HPV39 | HPV39-P1-F | CCTGTCTACCTCTATAGAGTCTTCCATACCTTCTAC | SEQ ID NO:17 |
| HPV45 | HPV45-P1 | TTATGTGCCTCTACACAAAATCCTGTGCC | SEQ ID NO:18 |
| HPV51 | HPV51-P1 | CTATTAGCACTGCCACTGCTGCAGTTTCC | SEQ ID NO:19 |
| HPV52 | HPV52-P1-F | CAGTTGTAGATACCACTCGTAGCACTAACATGACTTTA | SEQ ID NO:20 |
| HPV53 | HPV53-P1-F | CATGACTCTTTCTGCAACCACACAGTCTATG | SEQ ID NO:21 |
| HPV56 | HPV56-P1 | CTGCTACAGAACAGTTAAGTAAATATGATGCACG | SEQ ID NO:22 |
| HPV58 | HPV58-P1 | GATACCACTCGTAGCACTAATATGACATTATGCAC | SEQ ID NO:23 |
| HPV59 | HPV59-P1 | CAGCACCAATCTTTCTGTGTGTGCTTC | SEQ ID NO:24 |
| HPV66 | HPV66-P1-F | CTACCAGAAGTACCAACATGACTATTAATGCAGC | SEQ ID NO:25 |
| HPV68 | HPV68-A-P2-F | CGTTGTGGATACAACGCGCAGTACTAATT | SEQ ID NO:26 |
| HPV68 | HPV68-B-P2-F | TATTTCTTACTGTTGTGGATACCACTCGCAGTACC | SEQ ID NO:27 |
| HPV70 | HPV70-P1-F | ATTGTCTGCCTGCACCGAAACGG | SEQ ID NO:28 |
| HPV73 | HPV73-P1-F | GTGTAGGTACACAGGCTAGTAGCTCTACTACAACG | SEQ ID NO:29 |
| HPV82 | HPV82-P3-F | CACTGCTGTTACTCCATCTGTTGCACAA | SEQ ID NO:30 |
The interior label primer probe of table 3 and sequence
| Primed probe title | Sequence | Sequence number |
| E-Ex4-F1 | TCAGCAACAACCCTGCCCTG | SEQ ID NO:31 |
| E-Ex4-R1 | GAAGGTCTTGGTGGACCCGTC | SEQ ID NO:32 |
| E-Ex4-P1 | TGGGACGGGACACGTTGCACCTCT | SEQ ID NO:33 |
Another object of the present invention is to provide the kit of 19 kinds of high-risk HPVs of detection, it is characterised in that the reagent
Box includes sequence such as SEQ ID NO:1~SEQ ID NO:Primer shown in 10, and sequence such as SEQ ID NO:11~SEQ
ID NO:The one or more of 30 specific probe, and sequence such as SEQ ID NO:31~SEQ ID NO:It is interior shown in 33
Index thing and probe；
The kit uses noncompetitive internal standard detecting system, chooses one section of conservative region of human genome as interior
Mark amplification purpose fragment, probe marks VIC fluorescence, is used to monitor sample DNA whether there is correct add and PCR expansions as internal standard reagent
Increasing course of reaction has without exception.CY5 fluorophors, other 17 kinds of high-risk-types are marked for 16/18 high-risk HPV DNA probe
HPV DNA probe flag F AM fluorophors, internal standard probe mark VIC fluorophors.
The kit forms are as shown in table 4.
Internal standard testing result can be truly reflected sample DNA mass and experimental implementation situation, can be used as to reagent,
The Quality Control of DNA mass and experimental implementation, the amplification region of selection is human EGFR gene conservative section relatively, and target sequence is long
Spend about 100bp.
The kit forms of table 4
Another aspect of the present invention provides a kind of reaction system for being used to detect 19HPVs, specific as follows：
The application method of kit of the present invention comprises the following steps：
1. sample collection：Applicable specimen types have urogenital tract secretion cotton swab, genital tract scraping blade, tissue biopsy
Sample etc.；
2. extract sample DNA；
3. the DNA extracted is mixed according to correspondence ratio with mixed enzyme, and it is added in PCR reaction tubes；And by HPV19
Positive control and HPV19 negative controls are separately added into corresponding PCR reaction tubes；
4. the Ct values shown according to fluorescent PCR amplification instrument judge testing result；
5. the explanation of testing result, as shown in table 5：
1. HEX (or VIC) passage should have substantially or have the amplification curve of " S " type trend to rise, and Ct value≤29.If Ct values
>29 or HEX passages rise without obvious amplification curve, illustrate add DNA contain PCR inhibitor or DNA additions it is very few, it is necessary to
Again extract or increase DNA applied sample amounts and tested again；In such cases, if FAM the and CY5 passages of sample one of them or two
Individual rise and Ct value≤27, then the sample need not be detected again, as a result be judged as that high-risk HPV DNA is positive.
2. the amplification curve of CY5 passages is judged as the types of HPV 16/18 sun in typical " S " type and Ct value≤27, then result
Property.
1. the amplification curve of FAM passages is judged as other high-risk-types of HPV in typical " S " type and Ct value≤27, then result
It is positive.
2. a sample is probably that the types of HPV 16/18 and other high-risk-types are simultaneously positive.
The sample results of table 5 judge
The beneficial effects of the invention are as follows：
Molecular diagnostic techniques can then improve the sensitivity and specificity of HPV detections.By cytology detection method and molecule
Diagnostic method combines the diagnostic level that can preferably improve that cervical carcinoma and other anogenital cancers become.The present invention provides a kind of highly sensitive
The primer, probe and detection architecture that are used to detect the parting of HPV 19 of degree and high specific.The detection kit is adopted
Purpose fragment is expanded with a plurality of universal primer, amplified fragments detected in real time using special fluorescence probe, is grasped
Make that simple and efficient there is provided a kind of fast and reliable detection method.This method can realize that same reaction tube disposably detects 19 kinds
HPV (HPV), substantially reduces time and the examination expense of detection, for clinical early diagnosis provide it is important according to
According to.
The present invention can detect 19 kinds of high-risk HPVs in same reaction tube, i.e., the present invention uses general primer and pin
To various other Idiotype probe, the difference of the probe by adding Idiotype can constitute detection one or more, at most may be used
To detect 19 kinds of high-risk HPVs.The amplified fragments of selection are located at the L1 gene regions of HPV genomes, using a plurality of universal primer pair
Purpose fragment is expanded, and amplified fragments is detected in real time using special fluorescence probe, using noncompetitive internal standard
Detecting system, chooses one section of conservative region of human genome as amplification of internal standard purpose fragment, probe marks VIC fluorescence, as
Internal standard reagent is used to monitor sample DNA and whether there is correctly to add have without exception with pcr amplification reaction process.For 16/18 high-risk-type
HPV DNA probe mark CY5 fluorophors, other 17 kinds of high-risk HPV DNA probe flag F AM fluorophors, internal standard
Probe marks VIC fluorophors.Judge finally according to PCR amplification curves among system with the presence or absence of kit detection range
High-risk HPV DNA.Using universal primer, the complexity of system primer consumption and system is reduced, cost has been saved；10 are drawn
Thing collective effect detects all high-risk-types；19 specific probe makes a distinction to each type.PCR amplifications of the present invention are anti-
Answer system simultaneously contain UNG enzymes, can selectively be broken the uracil glycosidic bond in the PCR fragment containing dU, effectively reduce because
The false positive that PCR primer pollution is produced.
Sensitivity of the present invention is high (reaching 5 copies/reaction, improve more than 2 orders of magnitude than existing certain methods), specifically
Property it is strong, it is simple and efficient to handle, meanwhile, for patients, save examination expense.
Brief description of the drawings
Fig. 1 is HPV16 and HPV18 sensitivity analysis figures.
Fig. 2 is 17 kinds of HPV sensitivity analysis figures.
Fig. 3 is national full-length genome parting reference material (HPV16 and HPV18) detection figure.
Fig. 4 is national full-length genome parting reference material (HPV26/31/33/35/45/56) detection figure.
Fig. 5 is national full-length genome parting reference material (HPV58/59/66/6/73/82) detection figure.
Fig. 6 is national full-length genome parting reference material negative reference product N1-N5 detection figures.
Fig. 7 is national full-length genome parting reference material (HPV6/11/61/67/69/71/81) detection figure.
Fig. 8 is clinical negative sample testing result figure.
Fig. 9 is HPV16/18 positive clinical sample amplification figures.
Figure 10 is other high-risk-type positive clinical pattern detection result figures.
Embodiment
Below in conjunction with specific embodiment, the present invention is expanded on further.It should be understood that these embodiments be only used for the present invention and
It is not used in limitation the scope of the present invention.Unless otherwise defined or described herein, described in this patent scientific and technical term and this area is general
Logical technical staff is understood with identical implication.
Embodiment 1
The present embodiment using 19 kinds of high-risk human mammilla papillomavirus DNA plasmids in full-length genome parting reference material as template,
Real-time PCR detection is carried out using kit of the present invention：
(1) PCR reaction systems are set up
Using the probe and special primer of above-mentioned design, 5 μ L reference materials DNA are taken as reaction template, it is anti-using following PCR
System is answered to be expanded, pcr amplification reaction system is：
Having added the PCR reaction tubes of template to be put into real-time PCR detection instrument, carried out by following amplification program：First
Stage：50 DEG C of 2min, 95 DEG C of 5min, 1 circulation；Second stage：95 DEG C of 5s, 40 DEG C of 30s, 72 DEG C of 30s, 10 circulations；3rd
Stage：95 DEG C of 5s, 60 DEG C of 35s, 72 DEG C of 30s, 35 circulations；Phosphor collection：Collected during 60 DEG C of phase III FAM, HEX (or
) and CY5 signals VIC.
(2) the Ct values shown according to fluorescent PCR amplification instrument judge testing result, as shown in table 5.
1. HEX (or VIC) passage should have substantially or have the amplification curve of " S " type trend to rise, and Ct value≤29.If Ct values
>29 or HEX passages rise without obvious amplification curve, illustrate add DNA contain PCR inhibitor or DNA additions it is very few, it is necessary to
Again extract or increase DNA applied sample amounts and tested again；In such cases, if FAM the and CY5 passages of sample one of them or two
Individual rise and Ct value≤27, then the sample need not be detected again, as a result be judged as that high-risk HPV DNA is positive.
2. the amplification curve of CY5 passages is judged as the types of HPV 16/18 sun in typical " S " type and Ct value≤27, then result
Property.
3. the amplification curve of FAM passages is judged as other high-risk-types of HPV in typical " S " type and Ct value≤27, then result
It is positive.
4. a sample is probably that the types of HPV 16/18 and other high-risk-types are simultaneously positive.
Sensitivity analysis：Concentration is the reference material DNA of 500 copies after taking quantitatively, carries out 10 times of gradient dilutions, 3 are done altogether
Individual dilution gradient, 5 μ L, 8 parallel carry out amplified reactions are added per secondary response.As a result show, sensitivity of the present invention can be detected
To 5 copies/μ L.
Testing result shows that detection architecture of the invention can accurately detect 19 kinds of high-risk HPVs, and the sensitivity of detection can
Reach 5 copies/μ L.
Embodiment 2
Reaction system such as embodiment 1, is referred to using kit of the present invention to the national full-length genome parting of HPV
Each component in product is detected, investigates the reaction system finally determined and HPV full-length genome parting country is referred to
The detection case of product, result of the test such as Fig. 3-7.
Result of the test shows that this kit is to the negative reference in HPV full-length genome parting National reference
Product N1-N5 detection be feminine gender, in HPV full-length genome parting National reference 7 kinds of low risks (HPV6,
11st, 61,67,69,71,81) positive reference product are detected as feminine gender, to HPV full-length genome parting National reference
In 13 kinds of high-risk-types (HPV16,18,26,31,33,35,45,56,58,59,66,73,82) positive reference product examine survey be sun
Property, show that the reaction system finally determined is to the detection coincidence rate of HPV full-length genome parting National reference
100%.
Embodiment 3
Clinical sample is detected with the present invention, 80 clinical samples are detected altogether, and have with sub- energy biotechnology (Shenzhen)
The human papilloma virus kit for detecting nucleic acid (PCR- fluorescence probe methods) (17 type) of limit company is used as control.Utilize the present invention
The step of real-time fluorescence PCR system detects 80 clinical samples is as follows.
(1) detection sample DNA is extracted
Detect that sample includes urogenital tract secretion cotton swab, genital tract scraping blade, tissue biopsy specimen etc..
In centrifuge tube clean the sample transfer 1mL to one gathered, 12,000rpm centrifuge 1 minute；Carefully siphon away
700 μ L of supernatant liquid, 350 μ L Buffer VDL12 (plus absolute ethyl alcohol) are added into remaining precipitated liquid；Concussion mixes 10s, uses
Hand held 5~10s of centrifuge, is placed in 80 DEG C of digestion 3min in incubator；Concussion mixing 10s, hand held centrifuge 5~
10s, whole liquid are transferred in CB3DNA adsorption columns, 12000rpm centrifugations 30s；The liquid in collecting pipe is outwelled, is inhaled toward DNA
400 μ L Buffer RTW, 12000rpm centrifugation 3min (not empty to get rid of) are added in attached column；Collecting pipe is abandoned, adsorption column is carefully turned
Move in clean 1.5mL centrifuge tubes, 80 μ L Buffer BDE are added dropwise toward DNA absorption center membranes；It is stored at room temperature 2min,
12000rpm centrifuges 1min, collects sample DNA and preserves.
(2) PCR reaction systems are set up
Using the probe and special primer of above-mentioned design, 5 μ L reference materials DNA are taken as reaction template, it is anti-using following PCR
System is answered to be expanded, pcr amplification reaction system is：
Having added the PCR reaction tubes of template to be put into real-time PCR detection instrument, carried out by following amplification program：First
Stage：50 DEG C of 2min, 95 DEG C of 5min, 1 circulation；Second stage：95 DEG C of 5s, 40 DEG C of 30s, 72 DEG C of 30s, 10 circulations；3rd
Stage：95 DEG C of 5s, 60 DEG C of 35s, 72 DEG C of 30s, 35 circulations；Phosphor collection：Collected during 60 DEG C of phase III FAM, HEX (or
) and CY5 signals VIC.
(3) the Ct values shown according to fluorescent PCR amplification instrument judge testing result, as shown in table 5.
1. HEX (or VIC) passage should have substantially or have the amplification curve of " S " type trend to rise, and Ct value≤29.If Ct values
>29 or HEX passages rise without obvious amplification curve, illustrate add DNA contain PCR inhibitor or DNA additions it is very few, it is necessary to
Again extract or increase DNA applied sample amounts and tested again；In such cases, if FAM the and CY5 passages of sample one of them or two
Individual rise and Ct value≤27, then the sample need not be detected again, as a result be judged as that high-risk HPV DNA is positive.
2. the amplification curve of CY5 passages is judged as the types of HPV 16/18 sun in typical " S " type and Ct value≤27, then result
Property.
3. the amplification curve of FAM passages is judged as other high-risk-types of HPV in typical " S " type and Ct value≤27, then result
It is positive.
4. a sample is probably that the types of HPV 16/18 and other high-risk-types are simultaneously positive.
As a result show, 80 detected altogether clinical sample, contrast agents box detection positive sample 39, negative sample 41
Example, and this kit detection positive sample 43, negative sample 37；Compared with contrast agents box, the positive symbol of this kit
Conjunction rate is up to 92.3%, and negative match-rate is up to 97.6%, and total coincidence rate is up to 95%, and result of the test visualizingre agent box has been criticized with market
Quasi- kit has good coincidence rate.Such as Fig. 8-10.
Claims (3)
1. a kind of kit for being used to detect 19 kinds of high-risk human mammilla papillomavirus, it is characterised in that including：
1) primer
General HPV-F1TGCCTGGGGYAATCAGTTATTTGTTAC SEQ ID NO:1
General HPV-R3CCTCTGCCATGICKAATATATTSCTTAA SEQ ID NO:2
General HPV-R5CCTCTGCCATGYCKAATATAYTSCTTAA SEQ ID NO:3
General HPV-R6CCTCTGCCATGYCKAATATACTSCTTAA SEQ ID NO:4
HPV39 HPV-390-F1TGCCTGGGGCAATCAGTTATTTGTTAC SEQ ID NO:5
HPV53 HPV-530-R1CCTCTGCCATGTCTAAGGTATTCCTTAA SEQ ID NO:6
HPV51 HPV-510-R CCTCTGCCATGACTAACCTATTCCTTAA SEQ ID NO:7
HPV16 HPV-160-R CCTCTGCCATGYCTAATATAYTSCTTAA SEQ ID NO:8
HPV18 HPV-180-F TGCCTGGCATAATCAATTATTTGTTAC SEQ ID NO:9
HPV52 HPV-520-R CCTCTGCCATGTCTAATATATTCCTTAA SEQ ID NO:10；
2) probe
HPV16 HPV16-P1-C CGCAGTACAAATATGTCATTATGTGCTGCC SEQ ID NO:11
HPV18 HPV18-P3-C TTAACAATATGTGCTTCTACACAGTCTCCTGTACC SEQ ID NO:12
HPV26 HPV26-P1-F GTTGATACCACCCGCAGTACTAACCTTACC SEQ ID NO:13
HPV31 HPV31-P1-F CCAATATGTCTGTGTGTGCTGCAATTGC SEQ ID NO:14
HPV33 HPV33-P1 CCACTCGCAGTACTAATATGACTTTATGCACAC SEQ ID NO:15
HPV35 HPV35-P1-F GTCTGTGTGTTCTGCTGTGTCTTCTAGTGACAG SEQ ID NO:16
HPV39 HPV39-P1-F CCTGTCTACCTCTATAGAGTCTTCCATACCTTCTAC SEQ ID NO:17
HPV45 HPV45-P1 TTATGTGCCTCTACACAAAATCCTGTGCC SEQ ID NO:18
HPV51 HPV51-P1 CTATTAGCACTGCCACTGCTGCAGTTTCC SEQ ID NO:19
HPV52 HPV52-P1-F CAGTTGTAGATACCACTCGTAGCACTAACATGACTTTA SEQ ID NO:20
HPV53 HPV53-P1-F CATGACTCTTTCTGCAACCACACAGTCTATG SEQ ID NO:21
HPV56 HPV56-P1 CTGCTACAGAACAGTTAAGTAAATATGATGCACG SEQ ID NO:22
HPV58 HPV58-P1 GATACCACTCGTAGCACTAATATGACATTATGCAC SEQ ID NO:23
HPV59 HPV59-P1 CAGCACCAATCTTTCTGTGTGTGCTTC SEQ ID NO:24
HPV66 HPV66-P1-F CTACCAGAAGTACCAACATGACTATTAATGCAGC SEQ ID NO:25
HPV68 HPV68-A-P2-F CGTTGTGGATACAACGCGCAGTACTAATT SEQ ID NO:26
HPV68 HPV68-B-P2-F TATTTCTTACTGTTGTGGATACCACTCGCAGTACC SEQ ID NO:27
HPV70 HPV70-P1-F ATTGTCTGCCTGCACCGAAACGG SEQ ID NO:28
HPV73 HPV73-P1-F GTGTAGGTACACAGGCTAGTAGCTCTACTACAACG SEQ ID NO:29
HPV82 HPV82-P3-F CACTGCTGTTACTCCATCTGTTGCACAA SEQ ID NO:30
3) internal standard
E-Ex4-F1 TCAGCAACAACCCTGCCCTG SEQ ID NO:31
E-Ex4-R1 GAAGGTCTTGGTGGACCCGTC SEQ ID NO:32
E-Ex4-P1 TGGGACGGGACACGTTGCACCTCT SEQ ID NO:33。
2. a kind of kit for being used to detect 19 kinds of high-risk human mammilla papillomavirus as claimed in claim 1, it is characterised in that
Also include：
HPV19 mixed enzymes：Including Taq archaeal dna polymerases, the μ L of 1 pipe of UNG enzymes 27
HPV19 positive controls：Including the μ L of 18/26 type plasmids of HPV, 1 pipe 50.
3. a kind of reaction system for being used to detect 19 kinds of HPVs, specific as follows：
DEPC H2O
10 × PCR buffer solutions
MgCl2 20.0-50.0mmol
50-200 μm of ol of each pair of primer of claim 1
50-200 μm of ol of each pair of probe of claim 1
dNTP 20.0-30.0mmol
Mixed enzyme 0.5-1.0 μ L
Cumulative volume 30-50 μ L.
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| CN109554506A (en) * | 2016-01-15 | 2019-04-02 | 艾康生物技术(杭州)有限公司 | It is a kind of for detecting the design method of the primer and probe of pathogen |
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| CN107267663A (en) * | 2017-05-08 | 2017-10-20 | 常州金麦格生物技术有限公司 | HPV detection methods and kit |
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| US20110111389A1 (en) * | 2001-11-07 | 2011-05-12 | Diagcor Bioscience Incorporation Limited | Rapid genotyping analysis for human papillomavirus and the device thereof |
| GB0516145D0 (en) * | 2005-08-05 | 2005-09-14 | Genomica Sau | In vitro diagnostic kit for identification of human papillomavirus in clinical samples |
| NO330943B1 (en) * | 2009-04-30 | 2011-08-22 | Unilabs Telelabs As | A method for detecting and / or typing and / or quantifying human papillomavirus (HPV) type, primers and probes thereof, and diagnostic and use thereof. |
| CN103320533A (en) * | 2013-06-26 | 2013-09-25 | 清华大学 | HPV (human papillomavirus) DNA (deoxyribonucleic acid) genetic typing method and kit thereof |
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