CN104614524A - Test strip for rapid cervical cancer HPV detection device - Google Patents

Test strip for rapid cervical cancer HPV detection device Download PDF

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Publication number
CN104614524A
CN104614524A CN201510091653.XA CN201510091653A CN104614524A CN 104614524 A CN104614524 A CN 104614524A CN 201510091653 A CN201510091653 A CN 201510091653A CN 104614524 A CN104614524 A CN 104614524A
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China
Prior art keywords
pad
sample
nucleic acid
test strip
base plate
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CN201510091653.XA
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Chinese (zh)
Inventor
张鹭鹭
李先坤
李晓磊
程昀静
其他发明人请求不公开姓名
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Henan Kelong Medical Apparatus & Instruments Co Ltd
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Henan Kelong Medical Apparatus & Instruments Co Ltd
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G01N33/57411—Specifically defined cancers of cervix
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983—Viruses
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/70—Mechanisms involved in disease identification
    • G01N2800/7023—(Hyper)proliferation
    • G01N2800/7028—Cancer

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Virology (AREA)
  • Biotechnology (AREA)
  • Food Science & Technology (AREA)
  • Pathology (AREA)
  • Microbiology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Oncology (AREA)
  • Hospice & Palliative Care (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention provides a test strip for a rapid cervical cancer HPV detection device. The test strip comprises a bottom plate, a sample pad, a bonding pad and an absorption pad, wherein the sample pad is arranged above on end of the bottom plate; the absorption pad is arranged above the other end of the bottom plate; the bonding pad is overlapped below one end of the sample pad; a nitrocellulose membrane pad is arranged between the bonding pad and the absorption pad; a test line and a control line are arranged on the nitrocellulose membrane pad; and nucleic acid probes are respectively arranged on the test line and the control line. The test strip has the advantages that 1, the sensitivity is high, namely according to the design of the nucleic acid probes, the sensitivity of the detection method can be greatly improved during application; and 2, the specificity is high, compared with high nonspecific binding interference of antigens-antibodies, due to the design of the specific nucleic acid probes, the nonspecific action in a sample can be greatly reduced during actual application.

Description

A kind of test strips of cervical carcinoma HPV device for fast detecting
Technical field
The present invention relates to cervical carcinoma detection field, particularly relate to a kind of test strips of cervical carcinoma HPV device for fast detecting.
Background technology
HPV virus is a kind of double-stranded DNA virus, has spherical housing, diameter 55nm, and the mucocutaneous epithelium of main infection, causes different lesions.The HPV virus identified at present is more than 200 kinds, and at least 30 kinds are infected relevant to genital tract mucosa.HPV women 80% can infect HPV in life, is usually naturally removed in 8-10 month, only has minority (5%) women to be persistent infection state.
HPV persistent infection is the primary cause of disease causing cervical carcinoma.Cervical carcinoma is one of common gynecologic malignant tumor, and the incidence of disease occupies female malignant second.China is a cervical carcinoma district occurred frequently, about annual new cases 200,000 people, accounts for nearly 1/3 of the total number of the infected in the world, and about has 50,000 people to die from cervical carcinoma every year.Along with shifting to an earlier date of present young man's sexual life, early childbirth, sexual life is disorderly, immunity is low etc. increases again the risk suffering from cervical carcinoma and relevant disease to a great extent.How to prevent and treat and treat the important topic that cervical carcinoma has become current social development.
At present, the methods such as PCR, HCII, TCT are adopted more to the detection of HPV virus.But above-mentioned several method complex operation, detection time is long, accuracy rate is lower, need specific instrument and equipment to detect, need special technician's operation and testing cost is higher, so these factors are unfavorable for the early stage rapid screening of nucleic acid.Therefore, need a kind of method of quick detection pathogen nucleic acid newly, can the existence of direct-detection pathogen.Although there has been the multiple method determining microbial nucleic acids, but existing technology to be used as in diagnosis index still Shortcomings.Therefore, set up a kind of sensitive, fast nucleic acid detection technique be the problem that research worker inquires into always.
Summary of the invention
Object of the present invention for solve above technical matters, provide a kind of highly sensitive, stability is high, it is quick to detect, the test strips of easy to carry, free of contamination cervical carcinoma HPV device for fast detecting.
In order to solve the problems of the technologies described above, the concrete technical scheme taked is as follows: a kind of test strips of cervical carcinoma HPV device for fast detecting, comprise base plate, sample pad, pad, absorption pad, be sample pad above described base plate one end, be absorption pad above the base plate other end, below sample pad one end, overlap has pad, is nitrocellulose filter pad between pad and absorption pad, nitrocellulose filter pad is provided with p-wire and control line, p-wire and control line are respectively arranged with nucleic acid probe.
Further, described base plate is inert plastic.
Further, described pad is fiberglass packing, and fiberglass packing is coated with nano material, and nano material is golden nanometer particle and is made up of the silicon nanowires of Nanoparticle Modified.
The invention has the beneficial effects as follows, this test strips has the following advantages: 1. highly sensitive: the design of nucleic acid probe, makes it greatly can improve the sensitivity of detection method when applying.2. high specificity: the non-specific binding interference of antigen-antibody is compared more by force, and the design of special nucleic acid probe makes it can reduce non-specific effect in sample to greatest extent in actual applications.3. stability is high: in the present invention, mark be nucleic acid probe because the stability combined than antigen-antibody in the stability of extraneous nucleic acid hybridization is high.4. detect fast: result interpretation generally can complete in 20 minutes.5. easy to carry, simple to operate: to detect without any need for special instrument or equipment, operation steps is simple, and without the need to pcr amplification, layman can operate.Be convenient to medical personnel carry, also can for general population or patient oneself detecting & monitoring.Be convenient to rapid screening and the postoperative tracking treatment of HPV, be applicable to the grass-roots unit such as backcountry and the underdeveloped hospital of medical and health conditions.6. cost is low; 7. can carry out the detection of HPV somatotype fast; 8. this test strips is disposable product, environmentally safe, according to general treatment of!medical waste.
Accompanying drawing explanation
Fig. 1 is structural representation of the present invention;
Embodiment
In order to make object of the present invention, technical scheme and advantage clearly understand, below in conjunction with drawings and Examples, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
According to Fig. 1 as shown, a kind of test strips of cervical carcinoma HPV device for fast detecting, comprise base plate 6, sample pad 1, pad 2, absorption pad 5, be sample pad 1 above described base plate 6 one end, be absorption pad 5 above base plate 6 other end, below sample pad 1 one end, overlap has pad 2, it is nitrocellulose filter pad 8 between pad 2 and absorption pad 5, nitrocellulose filter pad 8 is provided with p-wire 3 and control line 4, p-wire 3 and control line 4 are respectively arranged with nucleic acid probe 7, base plate 6 is inert plastic, pad 2 is fiberglass packing, fiberglass packing is coated with nano material, nano material is golden nanometer particle and is made up of the silicon nanowires of Nanoparticle Modified.
Concrete principle of work is:
One, sample process
This test strips is applied to HPV rapid screening and monitoring after operation, applied widely, can be used for detecting the polytype samples such as paraffin-embedded tissue (FFPE), dipped into formalin tissue, blood sample, cervical exfoliated cell.Illustrate the disposal route of cervical exfoliated cell and FFPE two kinds of samples herein.
1, the process of cervical exfoliated cell:
A. outwell supernatant after sample being poured into 15mL centrifuge tube centrifugal (3000rpm), with again centrifugal after 2mLDDH2O cleaning, outwell supernatant; B. add following mixed liquor: 200 μ L lysates and 400 μ L DDH2O (blending ratio is 1:2), blow and beat cell mass with liquid-transfering gun, cell dispersal is suspended; C. add 5 μ L Proteinase Ks again, place 1 hour in 65 DEG C, 2 ~ 3 concussions in cracking process, can be carried out.D. supernatant is got for detecting.
2, the process of the sliced sample of FFPE:
A. get paraffin-embedded tissue section to be placed in 65 DEG C of baking ovens and to toast 20-30min, paraffin is tentatively melted; B. after paraffin tentatively melts, add successively in following solutions carry out dewaxing, aquation: dimethylbenzene Ι (10min), dimethylbenzene II (10min), absolute ethyl alcohol (2min), 50% ethanol (2min), washing; C. the biopsy tissues cover glass after washing is scraped in 1.5mL EP pipe gently, add 300 μ L Tissue lysates and 2 μ L Proteinase Ks, hatch 1-3h for 65 DEG C, every concussion in 1 hour once, until solution becomes limpid (as organized the more incubation time that extends to 3-5h; Understand the precipitations such as residual fraction connective tissue after sample cracking, do not affect subsequent experimental); D. directly get supernatant to detect.
The process of 3.FFPE tubular specimen:
A. observe paraffin organization sample, bulk paraffin in pipe can first be removed; B. dimethylbenzene washing: add 800 μ L dimethylbenzene, mixing is placed on 65 DEG C, 10min, accelerates the dissolving of paraffin; C.13000rpm centrifugal 3min.Careful absorption supernatant, keeps off bottom tissue precipitation (if any more paraffin, can repeat step 2,3, until paraffin is removed); D. ethanol aquation: add 800 μ L 100% ethanol, concussion mixing, 13000rpm, 5min.Carefully remove supernatant; E. 800 μ L 50% ethanol are added, concussion mixing, 14000rpm, 10min.Carefully remove supernatant, notice that keeping off bottom tissue precipitation (notes: not easily precipitate after organizing aquation, be different from dehydrated tissue and easily precipitate; As non-precipitate, centrifugation time can be increased to 15min.); F. add 500 μ L Tissue lysates and 5 μ L Proteinase Ks, hatch 1-3h for 65 DEG C, every concussion in 1 hour once, until solution becomes limpid (as organized the more incubation time that extends to 3-5h; Understand the precipitations such as residual fraction connective tissue after sample cracking, do not affect subsequent experimental); G. directly get supernatant to detect.
Two, ELISA test strip:
Get after the supernatant after 10 μ L sample cracking and 40 μ L DDH2O are mixed, add test strips well, after the complete standing 15min of application of sample, then add 30 μ L DDH2O and clean, after waiting for 5min, get final product observations.
Three, interpretation of result
If there is specific nucleic acid to be measured in sample, all visible colour band of the p-wire of test strips and control line.
If without specific nucleic acid to be measured in sample, the only visible colour band in the control line place of test strips.
If the p-wire of test strips and control line are all without visible colour band, illustrate that this testing result is invalid.

Claims (3)

1. the test strips of a cervical carcinoma HPV device for fast detecting, comprise base plate, sample pad, pad, absorption pad, it is characterized in that, be sample pad above described base plate one end, be absorption pad above the base plate other end, below sample pad one end, overlap has pad, is nitrocellulose filter pad between pad and absorption pad, nitrocellulose filter pad is provided with p-wire and control line, p-wire and control line are respectively arranged with nucleic acid probe.
2. the test strips of cervical carcinoma HPV device for fast detecting according to claim 1, is characterized in that, described base plate is inert plastic.
3. the test strips of cervical carcinoma HPV device for fast detecting according to claim 1, it is characterized in that, described pad is fiberglass packing, and fiberglass packing is coated with nano material, and nano material is golden nanometer particle and is made up of the silicon nanowires of Nanoparticle Modified.
CN201510091653.XA 2015-03-01 2015-03-01 Test strip for rapid cervical cancer HPV detection device Pending CN104614524A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106636307A (en) * 2015-11-02 2017-05-10 广东医学院 Application of miRNA-146a mutant in preparing test strip for early screening and rapid diagnosis of cervical cancer
CN109001467A (en) * 2018-07-20 2018-12-14 贵州省人民医院 A kind of gene association prognosis detection reagent and preparation method thereof for prostate cancer
WO2019239430A1 (en) * 2018-06-13 2019-12-19 Rajeshwar Nath Sharan Biomonitoring device for detection and monitoring of cancer and a method thereof

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106636307A (en) * 2015-11-02 2017-05-10 广东医学院 Application of miRNA-146a mutant in preparing test strip for early screening and rapid diagnosis of cervical cancer
WO2019239430A1 (en) * 2018-06-13 2019-12-19 Rajeshwar Nath Sharan Biomonitoring device for detection and monitoring of cancer and a method thereof
CN109001467A (en) * 2018-07-20 2018-12-14 贵州省人民医院 A kind of gene association prognosis detection reagent and preparation method thereof for prostate cancer

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Application publication date: 20150513