73 results about "High risk hpv" patented technology

The invention provides an sgRNA combined with immunogene to inhibit high risk HPV expression, a gene knockout vector thereof, and an application of the sgRNA and the vector. The sgRNA sequences of a human papilloma virus 16 type gene suitable for CRISPR-Cas9 targeting editing and a human PD-1 gene are selected, and the plasmid vector of the sgRNA and a CRISPR-Cas9 nuclease gene expression vector are transferred to HPV16+ human cervical carcinoma cells and HPV16+ transplantation tumor-bearing mice, and can obviously inhibit HPV16 expression and inhibit tumor growth. The gene expression vector prepared in the invention has the advantages of simple method steps, good sgRNA targeting property, high CRISPR-Cas9 knockout efficiency, accurate targeting splicing of high risk HPV16 type and PD-1 genes, efficient reduction of the expression of the high risk HPV16 type gene, and obvious inhibition of the tumor growth through combined application.

The invention discloses a primer, a probe and a kit for fluorescence PCR (Polymerase Chain Reaction) detection of 18 types of high-risk human papilloma viruses. Different typing kits are detected by adding different specificity probes; the DNAs (Deoxyribose Nucleic Acids)) of 18 types of common high-risk human papilloma viruses internationally recognized and closely related to the cervical cancer can be detected once; and typing detection is carried out on HPV (Human Papilloma Virus)16 and 18. The application provides 6 universal primers and 18 specific molecular beacon probes. The DNAs of the 18 types of common high-risk human papilloma viruses can be amplified by the 6 universal primers; and meanwhile, the 18 probes are the specific molecular beacon probes designed by aiming at the 18 high-risk types; different types of probes are added according to the detection need and combined as the kits aiming at the detection need of the different high-risk types; and at the same time, the added probes are marked by different report genes, so that the purpose of carrying out typing detection on the high-risk HPV is achieved.

The invention belongs to the technical field of medicines, and discloses a gel and a use thereof. The gel comprises a chitosan polymer, a hyaluronic acid matter, a lactic acid matter, a gel matrix, a wetting agent and a diluter. The gel has the advantages of good antibacterial effect, wide antibacterial spectrum (comprising high risk HPV (Human Papilloma Virus), physical antibacterial effect, no tolerance, biodegradability, safety and environment-friendliness and lasting effect for a long time, and has the functions of nourishing, lubricating, whitening, repairing skins or mucosal tissues, and removing spots and inhibiting cicatrisation for skins or mucosal tissues.

The present invention provides a method and a kit for determining whether a head and neck cancer is HPV-related. In one embodiment, an RNAscope® HPV assay was designed to detect the presence of E6 / E7 mRNA of certain high-risk HPV subtypes related to head and neck cancer. The present invention also provides a method and a kit for determining whether a cervical lesion is a benign lesion or a cervical intraepithethial neoplasm lesion. The present invention further provides a method for determining the progression of cervical intraepithethial neoplasm based on the spatial pattern and levels of the E6 / E7 mRNA of certain high-risk HPV subtypes. The present invention also provides a method for determining the risk of developing cervical cancer in a human diagnosed with cervical intraepithethial neoplasm based on presence and absence of the certain subgroups of high-risk HPV subtypes.

The invention relates to the pharmaceutical application of a medicament combination containing oil of zedoary turmeric. The medicament combination has new pharmaceutical application for preventing and treating various diseases, such as various HPV virus subtype infections, intraepithelial lesion(isl), papilloma and cervical carcinoma, and the like, in particular to high-risk HPV subtype infections. The medicament is prepared into various conventional formulations in the category of pharmaceutics, and the medicament combination can significantly prevent and treat multiple related diseases of HPV infection by clinical verification.

A testing and typing method and its reagent boxes for Human Papillomavirus(HPV), relating to exciters and nucleic acid probes which are especially for testing and typing of HPV. The invention fixes three types of special probes including different subgroups of low-hazard HPV and high-hazard HPV, subgroups with extra-low infection rate of low-hazard or high-hazard HPV, onto the same position or mediator. The invention also fixes oligomeric nucleoside probes of special HPV onto microballoons based on Luminex xMAP technical platform, which forms microballoons probes for testing HPV subgroups and typing slugs of HPV. A testing and typing method with simplicity of operator and its reagent boxes are invented. The method has much strongpoint including high-sensitivity, high-flux, stable result and good repeatability.

The invention relates to methods of categorizing a cervical tissue or cytology sample by performing an in situ hybridization assay using an antisense E6 or E7 probe on a cervical tissue sample, wherein the antisense E6 or E7 probe can simultaneously detect HPV DNA and HPV RNA; detecting the presence of HPV nucleic acid; and categorizing the cervical tissue sample based on HPV nucleic acid expression.

The invention provides a monoclonal rabbit antibody for identifying HPV16 positive cervical tissues. The antibody can be used for specifically detecting a biomarker HPV16E7 protein in tissues including cervical cancer and cervical lesions, so that HPV persistent infection related cervical cancer tissues from abnormal or noncancerous cervical epithelial tissues, cervical cancer caused by high-risk HPV infection can be accurately diagnosed, and risk early-warning can be especially performed for whether early cervical lesions cancerate or not, so that the misdiagnosed rate of cervical lesions can be effectively reduced, and injuries and resource waste caused by over treatment for patients can be improved.

The invention provides a method, oligonucleotide and kit for detecting common high-risk HPV (human papilloma viruses). A fluorescence PCR (polymerase chain reaction) technology is adopted, HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 26, 53, 66, 73 and 82 which possibly exist in a sample are subjected to initial detection and type identification. By the method, oligonucleotide and kit for detecting the common high-risk HPV, 18 high-risk types can be detected simultaneously, and the HPV 16 and 18 can be subjected to genetic typing simultaneously. On the basis of 18 types of high-risk HPV and affinity of other types on a phylogenetic tree, a high-risk HPV primer and a probe are designed, a background fluorescence value is reduced remarkably while detection accuracy, sensitivity and specificity are guaranteed, detection flux is increased remarkably, and detection cost is greatly reduced.

The invention discloses a nucleic acid detection kit for 12+2 high-risk human papilloma virus (HPV), comprising: (1) probes as shown in SEQ ID No:1-12 and primers as shown in SEQ ID No: 13-36 of a nucleotide sequence which is only complementary with the DNAs of 12 high-risk HPV; (2) a probe as shown in SEQ ID No:37 and primers as shown in SEQ ID No: 38-39 of a nucleotide sequence which is only complementary with the DNA of high-risk HPV16; (3) a probe as shown in SEQ ID No:40 and primers as shown in SEQ ID No: 41-42 of a nucleotide sequence which is only complementary with the DNA of high-risk HPV 18; and (4) a probe as shown in SEQ ID No: 43 and primers as shown in SEQ ID No: 44-45 of a nucleotide sequence which is only complementary with the DNA of an intracellular beta-globin gene, wherein each group of probes respectively mark different dyes with the maximum emission wavelengths of 518nm, 538-553nm, 574-575nm, 607-615nm, 640nm, 666-667nm and 690nm. The kit disclosed by the invention can furthest lighten huge cost brought by HPV gene detection in China, and the health of people is protected.

The present invention relates to a method for discrimination of p16INK4a overexpressing metaplasias from neoplastic or preneoplastic p16INK4a overexpressing lesions by determination of the level of high risk HPV encoded gene-products such as e.g. HPV E2 and / or HPV E7 molecules in biological samples in the course of cytological testing procedures. The method thus enables for reduction of false positive results in the p16INK4a based detection of anogenital lesions in cytological testing procedures.

The invention discloses a detection kit, detection system and detection method for 19 high-risk HPVs. The 19 high-risk HPVs comprise HPV16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 70, 73 and 82. According to the invention, a plurality of general primers are employed for amplification of target fragments, and specific fluorescence probes are used for real-time detection of the amplified fragments; since the general primers are employed, the primer amount of the system and complexity of the system are reduced, cost is saved, and all the high risk types are detected through combined action of 10 primers; and 20 specific fluorescence probes are used to distinguish different types. With the detection kit, detection system and detection method provided by the invention, the 19 high-risk HPVs can be detected in a reaction tube; high sensitivity and good specificity are obtained; operation is simple and fast; and for patients, screening cost is saved.

The invention relates to a kit for auxiliary diagnosis of cervical carcinoma and high-grade cervical intraepithelial lesion, and discloses a kit for detecting high-risk HPV. The kit for detecting the high-risk HPV comprises a high-risk HPV nucleic acid fluorescent PCR detection mixture and Taq enzyme. The invention also discloses a method for preparing the kit for detecting the high-risk HPV and a method for using the same. The kit can be used for detecting the high-risk HPV and overcomes the defects of high false positive rate, low specificity, high cost and the like of typing detection of the high-risk HPV in the prior art.

The present invention provides CD4+ T-cell epitopes in E6, E7 and E2 proteins from various strains of human papillomavirus (HPV). In some preferred embodiments, the present invention provides means for the development of HPV vaccines, in particular multivalent vaccines for the prevention of infection with high-risk HPV strains. In additional embodiments, the present invention provides means for the development of therapeutic vaccines against high-risk HPV types that prevent the development of benign and / or malignant tumors in infected individuals. The present invention further provides epitopes suitable for use in prophylactic and therapeutic vaccines.

The invention discloses a gene chip for detecting human papillomavirus (HPV), comprising a solid carrier and a human papillomavirus detecting probes fixed on the solid carrier. The detecting probes comprise nucleotide sequences in SEQ ID No.9-37, and the 27 types of HVP subtype specific probes in the invention correspondingly comprise 22 kinds of high-risk and mediate-risk HPV, and 5 kinds of low-risk HPV and two kinds of high-risk HPV integration (HPV16 / HPV18). The invention also discloses a detecting kit for detecting various subtypes of HPV in a sample with high speed and high flux, and the kit comprises the gene chip, a primer sequence for detection, a human GAPDH gene, special HPV lysing solution, PCR reaction liquid and a color reagent, wherein the detecting probe fixed on the gene chip has the nucleotide sequence in the SEQ ID No.9-37; the primer sequence for detection is used for amplifying the DNA sequence of the HPV in a clinical sample, and the human GAPDH gene is used for amplifying the clinical sample. The HPV detection has important significance for the clinical diagnosis of HPV as well as the early prevention, the diagnosis, the treatment and the postoperation tracking of cervical cancer.

The invention relates to the field of gene therapy and discloses a fixed-point knockout system for high-risk HPV (human papilloma virus) E6E7 gene, namely targeted cutting of TALENs (transcription activator-like effector nucleases) of HPV E6E7 gene, wherein a TALENs expression vector of the target specific site is designed according to the high-risk HPV E6E7 gene sequence, and the HPV E6E7 gene sequence in the cells are cut in a targeted manner so as to knock out the target gene and induce the apoptosis increase and proliferation inhibition of corresponding subtype cells while the TALENs do not act on the HPV positive or HPV negative cells of other subtypes. By transfecting the TALEN expression vector in a subcutaneous tumor model of HPV positive cervical cancer cells, the growth of subcutaneous tumor can be obviously inhibited, and the tumor weight is reduced. According to the method for targeted knockout of high-risk HPV E6E7 gene using TALEN provided by the invention, the virus oncogene can be efficiently broken on the DNA level so as to cause apoptosis and proliferation inhibition of the HPV infected cells, and the method is of tremendous therapeutic value on HPV infection related diseases and particularly precancerous lesions such as cervical intraepithelial neoplasias.

The invention relates to a kit for detecting high-risk human papillomavirus E6 / E7 mRNA, and discloses a detection method of the kit. The method comprises: 1, performing reverse transcription by usingrandom primers; and 2, detecting 14 kinds of high-risk HPV E6 / E7 mRNAs and an internal reference gene in a single PCR reaction. According to the present invention, with the primers and the probes designed for the 14 kinds of the high-risk HPVs, the high-risk HPV E6 / E7 mRNA can be specifically and rapidly detected, wherein the detection time is only 2.5 h; and the kit has characteristics of rapidness, high efficiency, high sensitivity, good specificity, real-time detection analysis and the like, provides the rapid and accurate molecular detection method for the general investigation of HPV andthe prevention and treatment of cervical cancer, has the greatly-reduced inspection cost, and further has important promotion and application value.

The invention provides a kit for fluorescent PCR (Polymerase Chain Reaction) detection of a high-risk HPV (Human Papilloma Virus). The kit comprises a nucleic acid releasing agent and a PCR reaction solution, wherein the nucleic acid releasing agent comprises 0.01-05mmol / L surfactin, 20-300mmol / L potassium chloride, 0.01-2% sodium dodecyl sulfate and 0.05-1% ethanol; the PCR reaction solution comprises high-risk HPV16-type and 18-type primer probe sequences and internal standard primer probe sequences. The invention aims at providing a kit for fluorescent quantitative PCR detection of high-risk Human Papilloma Virus, and the kit has the advantages of fastness in operation, simple method, high detection sensitivity and wide detection range. By virtue of the kit, a common fast fluorescent PCR detection is carried out on DNA nucleic acid fragments of the high-risk human papilloma virus in unknown samples, such as cast-off cells of the surface of a verruca acuminata body, women cervical epithelial cells and genital secretion and the detection results can be applied in the auxiliary diagnosis of the high-risk human papilloma virus infection and the early screening of cervical cancer.

The invention provides an HPV (human papilloma virus) high-risk typing fluorescence PCR (polymerase chain reaction) detection kit which comprises a nucleic acid releaser and two or more PCR reaction solutions, wherein the nucleic acid releaser comprises 0.01-0.5 mmol / L surfactin, 20-300 mmol / L potassium chloride, 0.01-2% of sodium dodecylsulfate and 0.05-1% of ethanol. One of the PCR reaction solutions comprises high-risk HPV 16 type and / or 18 type primer probe sequences, and another PCR reaction solution comprises an internal reference primer probe sequence. The kit provided by can be used for carrying out quick fluorescence PCR typing detection on DNA (deoxyribonucleic acid) nucleic acid segments of high-risk HPV in wart surface shedding cells, female cervix epithelial cells, genital secretion and other unknown samples; and the detection result can be used for auxiliary diagnosis of high-risk HPV infection, early screening of cervical carcinoma, follow-up survey of cervix pathological changes and instruction of vaccine development.

The invention discloses gynecological gel for preventing and inhibiting HPV viruses. The gynecological gel comprises sodium hyaluronate, extract of bitter bean, extract of argyle leaf, extract of wildchrysanthemum, extract of radix isatidis, extract of Astragalus mongholicus, extract of coptis root, mannitol, Carbomer, triethanolamine and water. The gynecological gel for preventing and inhibitingHPV viruses contains extract of multiple types of plants, does not stimulate the vagina, is safe and has no side effect. The sodium hyaluronate in the gel carries antiviral and antibacterial components which can fast penetrate into the skin and prevents mucous cells from being damaged by viruses. The gynecological gel can fast repair the skin tissue and improve the wound healing and regenerativecapacity. The extract of multiple types of plants in the gel can improve immunity and produce good antiviral effects. The experiment result shows that the gynecological gel has an effective rate of 91.1% to the low-risk HPV infection and an effective rate of 86.1% to the high-risk HPV infection.

The invention relates to application of 5-aminolevulinic acid or pharmaceutically acceptable salts thereof or a medicament composition containing the 5-aminolevulinic acid or the pharmaceutically acceptable salts thereof in preparing medicaments for treating HPV (Human Papilloma Virus) infection, which is characterized by being applied to treating patients with continuous high-risk HPV infection with viral load not less than 10. When in treatment implementation, the invention is very safe and has no haemorrhage or anesthesia in the treatment process, pathological tissues can be selectively damaged, normal cervical tissue structures a re reserved, and the invention has no influence on the reproductive function of the patient, can be used for repetitive treatment, and has fast drug metabolism, no phototoxicity and no need of being kept in the dark.

The present invention is high risk human papilomavirus (HPV) detecting process, which includes the following steps: 1. designing and synthesizing proper specific primer, and amplifying the target DNA of the detected sample specifically for the second time by means of one-step nest PCR technology; and 2. hybridizing and selectively typing the target DNA of the detected sample through twice specific selective amplification and the specific segment of high risk HPV by means of combination to low density DNA chip technology, and signal identification through fluorescent detecting or direct color developing. The present invention provides also one corresponding kit. The present invention can amplify specific target DNA segments of 13 kinds of HPV simultaneously and detect several kinds of virus subtypes of HPV through once reaction, and has the advantages of high speed, high efficiency, high sensitivity, high specificity, etc.

The invention provides a biological composition dressing for preventing and treating HPV. The biological composition dressing comprises PHPV protein, carbomer, glycerol, green tea extract and Yujie pure MP components. The biological composition dressing can be prepared in dosage forms of dressing suppository, gel dressing, lotion, film agent and the like. The biological composition dressing has an obvious effect on treating genital tract high-risk HPV infection and lower-level cervical lesions caused by high-risk HPV infection, blocking genital tract HPV infection, improving vaginal micro-environment, relieving vaginal loosening and drying symptoms, reducing vaginal inflammatory reaction and lowering the vaginal PH value.

The invention provides a high-risk human papilloma virus detection kit which comprises a HPV amplification primer. Universal fluorescent mark primers are introduced into a multiplex PCR (polymerase chain reaction) system, and more than ten amplicons different in length or fluorescence can be obtained by amplification by designing three fluorescent mark primers; a specific joint of 20bp is introduced to a 5' terminal of a specific amplification primer, the universal fluorescent mark primers are introduced into the PCR system, length of fluorescent marked amplicons obtained through multiplex PCR amplification is larger than that of a target amplification area by two joints, and a multiplex PCR product is applied to capillary electrophoresis analysis to obtain genotype. By the high-risk human papilloma virus detection kit, expense for fluorescent marking of multiplex PCR is further lowered, and the problems of increase of HPV subtypes and linear increase of detection cost are solved. Taking using a method to detect 14 high-risk HPV subtypes as an example, multiplex PCR cost can be lowered by 50%, and cost reduction amplitude is higher as detected HPV subtypes are increased.

Relating to the field of gene therapy, the invention provides a method for knockout of a high-risk human papillomavirus (HPV) E6E7 gene by zinc finger nucleases (ZFNs). According to a high-risk type HPV E6E7 gene sequence, the method designs ZFNs expression vectors of a targeted specific site to targetedly cut the HPVE6E7 gene sequence in cells, thereby knocking out a target gene. In corresponding subtype HPV infected cells, the ZFNs can significantly induce cell apoptosis and proliferation inhibition, and has no effect on other subtype HPV positive cells and HPV negative cells. Transfection of the ZFNs expression vectors in a subcutaneous tumor model of HPV positive cervical cancer cells can obviously inhibit the growth rate of subcutaneous tumors and alleviate tumors. The method for targeted knockout of the high-risk HPV E6E7 gene by ZFNs can efficiently destroy virus oncogenes at the DNA level so as to make HPV infected cells undergo apoptosis and proliferation inhibition, and has enormous treatment value to HPV infection related diseases, especially cervical intraepithelial neoplasias and other precancerous lesions.

The invention discloses a composition for detecting and typing high-risk HPV, a kit and a method and relates to the field of molecule biology detection, in particular to the field of HPV detection. The composition can be used for specifically detecting E6 / E7 mRNA of HPV types 16, 18, 31, 33, 35, 39, 45 ,51, 52, 53, 56, 58, 59, 66 and 68. At the same time, the invention also provides application ofthe composition in the detection of HPV E6 / E7 mRNA, the kit comprising the composition and the method for detecting the HPV E6 / E7 mRNA.

The invention relates to a primer and probe set and a kit for detecting a virus, in particular to a primer and probe set and a kit for detecting oncogene E6 / E7 mRNA typing of high-risk human papillomaviruses, and belongs to the biochemical technology field. Specific upstream and downstream primers and corresponding specific molecular beacon probes are designed ingeniously for oncogenes E6 / E7 of thirteen high-risk HPVs (human papilloma viruses) and reference genes beta-actin, and specific amplification and high-specificity detection of the thirteen high-risk HPVs and the reference genes beta-actin are realized. The primer and probe set and the kit can be applied to real-time quantification and typing detection of oncogene E6 / E7 mRNA of the HPVs, and can be also applied to nature determination and typing detection of the thirteen HPVs at PCR (polymerase chain reaction) end points by the molecular beacon probes. The thirteen HPVs refer to HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59and 68. The oncogene E6 / E7 mRNA of the HPVs serves as a detection target, so that false positive results and false negative results both caused by HPV L1 DNA serving as a detection method in the prior art can be effectively avoided; the applied molecular beacon probes have the advantages of high detection sensitivity, high specificity, simplicity in operation and the like.

The invention belongs to the technical field of biology, and discloses a primer and molecular beacon combination for quickly detecting human papillomaviruses (HPVs) based on an isothermal amplification technology, a kit and a detecting method. The primer and molecular beacon combination is shown as SEQ ID No.1-17, and can specifically recognize the 13 different types of high-risk HPVs and two types of medium-risk genotypes with the high incidence in Chinese population; and according to the kit and the method for real-timely detecting the HPVs through fluorescence based on the isothermal amplification technology, the 15 types of medium-and-high-risk HPVs can be detected within the short time (1.5 h), accuracy is good, sensitivity is high, repeatability is high, specificity is good, no nonspecific reaction with clinically similar pathogens exists, using is convenient, only an extracted template sample needs to be added into a reaction tube, the manual operation time is less than 5 min, the requirement of isothermal amplification for temperature control over an instrument is not high, and thus the primer and molecular beacon combination, the kit and the detecting method can be appliedto instant detection well.

The invention discloses a method for preparing a nano-micro-spherical type human papilloma virus resistant and gynecologic inflammation pathogene polyclone combined IgY antibody. The method comprisesthe following steps: A, screening combined pathogene with a high-risk HPV and gynecologic inflammation pathogene; B, introducing the combined pathogene into engineering bacteria, and expressing combined antiogen by using the engineering bacteria; C, injecting the combined antiogen into livestock so as to obtain an egg with a polyclone yolk immune globulin combined IgY antibody; D, separating and purifying a combined IgY antibody extract of the combined IgY antibody for preventing the HPV and the gynecologic inflammation pathogene from the egg; e, concentrating the combined IgY antibody extract, and performing freezing drying, thereby obtaining the nano-micro-spherical type human papilloma virus resistant and gynecologic inflammation pathogene polyclone combined IgY antibody. The polyclonecombined IgY antibody prepared according to the method has the characteristics of high activity, good stability, good specificity, high safety and good permeability, and can be applied to treatment and auxiliary treatment on HPV infection and mixed infection of multiple gynecologic viruses and pathogenic bacteria.