106results about How to "High sensitivity and specificity" patented technology

The present invention provides a method for detecting a gene mutation, comprising the step of performing PCR using generic PCR primers together with a blocking primer which competes with the generic primers and was modified at one end, and a method of diagnosing gene mutation-related diseases using the same. According to the invention, detection sensitivity and specificity can be increased by blocking the amplification of normal DNA and selectively amplifying mutant DNA.

The invention discloses an implicit malachite green hapten which retains the characteristic structures of the implicit malachite green, active arms which have different electronic structural properties and different carbon chain length and is suitable for being coupled with macromolecule carriers derive from different chemical sites of the structure of the implicit malachite green, the active arm is R group, aromatic amine, terminal carboxyl group linear chain alkyl ether or amido link with the structure of linear chain terminal carboxyl group; an implicit malachite green artificial antibody is prepared and obtained from implicit malachite green hapten and when being used for detecting the residual quantity of the implicit malachite green and malachite green, the antibody has high specificity and sensitivity and high accuracy, the recovery rate can reach 80 to 110 percent, at the same time, the operation is simple and fast, no complicated pretreatment process is needed, a great amount of samples can be detected at one time, the cost is low, the operation has little requirements to operators so as to be convenient for on-site monitoring and be complementary to conventional methods, therefore, a sensitive, fast, efficient and economical implicit malachite green hapten immunity detecting method is successfully set up.

The invention relates to a fluorescent silver cluster, and a preparation method and application thereof. The average particle size of the fluorescent silver cluster is 1nm, and the fluorescent silver cluster can emit fluorescence, of which the wavelength is 650nm or so, under the excitation of light with different wavelengths. The preparation method comprises the following steps: reacting a silver salt and a protective agent in a water solution to generate a soluble silver complex, and slowly adding a reducer to generate the water-soluble fluorescent silver cluster. The fluorescent silver cluster can be used as a reagent for detecting mercury ions. The high-stability nano silver cluster, which can emit strong red fluorescence within the range of quantum scale, is formed by using common and cheap chemical reagents through a simple and practical synthesis technique. The nano silver cluster has very high sensitivity and specificity when being used as a reagent for detecting metallic ions, especially mercury ions. The invention can be widely used in the technical fields of environment detection, food safety and the like.

The invention discloses a bacterium drug-resistant gene detection method, a gene chip and a kit. The kit comprises a primer pair for amplifying the bacterium drug-resistant gene to be detected, and detection probes. The gene chip comprises probes for detecting the bacterium drug-resistant gene to be detected. The method for detecting the drug-resistant gene by using the kit and gene chip comprises the following steps: 1) carrying out PCR (polymerase chain reaction) amplification on the drug-resistant gene in the sample DNA (deoxyribonucleic acid) to be detected by using the primers in the kit; 2) carrying out fluorescence labeling reaction on the amplified product and the detection probes in the kit; and 3) hybridizing the fluorescence-labeled amplified product with the gene chip to determine the drug-resistant gene contained in the sample. The detection method disclosed by the invention covers the detection of the most common 16 drug-resistant genes in clinic, and is beneficial to implementing rapid diagnosis of clinical bacterium drug resistance conditions, thereby providing supports for reasonably selecting therapeutic drugs in clinic.

The invention provides a telomerase activity detecting probe, a telomerase activity detecting reagent kit and a telomerase activity detecting method. The design of telomerase combination primers (TS) and reverse primers is simple, and TS only needs to comprise DNA (deoxyribonucleic acid) enzyme sequences and restriction enzyme cutting sites of cutting enzyme and does not need any modification; the reverse primers only need to comprise sequences complemented with multiple-G sequences extending from telomerase, so the primers are easy to design, and the feasibility is high. The telomerase activity detecting method provided by the invention has the advantages that the reverse primers are only combined with TS primers extending from result telomerase, the nonspecific amplification is reduced, in addition, in the chemical luminous reaction, only amplified telomerase multiple-G sequences and DNA enzyme can be combined with hemin to form a tetramer, and chemical luminous signals are generated, so the telomerase activity detecting method provided by the invention has higher specificity and higher sensitivity. In addition, the telomerase activity detecting reagent kit provided by the invention has the advantages that the cost is low, the operation is simple, and the time is saved.

The invention discloses an LAMP detection primer composition for phytophthora infestans and an LAMP detection kit and an LAMP detection method of the LAMP detection primer composition. The LAMP detection primer composition consists of a positive inner primer FIP, a reverse inner primer BIP, a positive outer primer F3, a reverse outer primer B3 and a reverse ring primer LB. The detection method disclosed by the invention is high in accuracy, high in specificity, convenient to operate and high in practicability, realizes constant-temperature multiplication, also provides a new technical platform for detection on the phytophthora infestans and can be used for performing high-sensitivity quick detection on the phytophthora infestans; meanwhile, pathogens can be identified at the initial stage of disease invasion, and the pathogens in field soil can be detected. The LAMP detection primer composition has an important significance for preventing and treating late blight of potatoes and tomatoes, reducing the blind use of pesticide, lowering the production cost and alleviating the environmental pollution of the pesticide.

The invention belongs to the technical field of medical care, and particularly provides a production method of an early pregnancy detection kit using urine as a detection sample. The production methodadopts the principle of a colloidal gold immunochromatography technology and uses a double-antibody sandwich method technology, a T1 detection line of a nitrocellulose membrane is coated with an anti-beta-HCG core fragment monoclonal antibody, a T2 detection line of the nitrocellulose membrane is coated with an anti-alpha-HCG monoclonal antibody, and a quality control line of the nitrocellulose membrane is coated with a goat anti-mouse IgG polyclonal antibody. The kit having a high sensitivity, a high specificity and a good stability and being able to rapidly detect the pregnancy or not and the ectopic pregnancy or not is obtained by controlling a series of production process steps and process parameters. The invention further provides the product obtained through the production method, and a use of the product.

The invention discloses a diagnostic reagent kit for trichinosis by employing dot-immunogold filtration assay. The invention has the following advantages: gold labeled SPA is taken as an antibody labeling tag, isopropyl beta-D-thiogalactoside is used for inducing the pMAL-c2X-Ts21/TB1 recombinant strain, expressing and purifying trichina Ts21 gene-encoded protein (Ts21 antigenic gene, ORF17.20, the access number at GenBank is U88239), after being purified on a nitrocellulose membrane (NCM), the Ts21 recombinant protein coats the detection point T, and human IgG coats the quality control point C, thus constructing Ts21 recombinant protein-dot-immunogold filtration assay (Ts21-DIGFA) and assembling the rapid diagnostic reagent kit for trichina with Ts21 recombinant protein by employing dot-immunogold filtration assay. The kit is simple to operate and has high sensitivity and specificity and good stability and repeatability; the detection results can be observed and judged within 1min immediately after adding samples, the diagnosis results are accurate and clear and misjudgment can not happen; and the kit is suitable for rapid diagnosis and serum epidemiological survey at the scene and is easy to popularize and apply extensively.

This invention relates to garlic virus RT-PCR test reagent box and its test method specially used in the test of GMV and GCLV, among which, the box includes: an inverse transcription reagent box, a PCR box and a positive control. The test method includes reaction of the inverse transcription, PCR reaction and electrophoresis detection, the electrophoresis containing GMV test result is the amplified 32Tbp strip and the result with GCLV is the amplified 469bp strip. The PCR part of the reagent box adopts small reaction system.

The invention discloses a human echinococcosis colloidal gold immunochromatographic assay urine testing quick diagnosis test paper card. The test paper card comprises a plastic card box and a detection module in the card box; the plastic card box is provided a sample loading hole and an observation window; the detection module comprises sample filtering paper, a nitric acid fibrous membrane and absorbent paper; echinococcosis resistance fluid antigen humanized single chain variable region antibody which is marked by colloidal gold is absorbed on the absorbent paper and nitric acid fibrous membrane; a quality control line and a detection line are marked on the echinococcosis resistance fluid antigen humanized single chain variable region antibody marked by the colloidal gold on nitric acid fibrous membrane; the sample filtering paper is opposite to the sample loading hole; and the nitric acid fibrous membrane is opposite to the observation window. The test paper card has the advantages of simplicity and convenience, fastness, no invasiveness, reliability and stability.

The invention relates to a warm starting TaqDNA polymerase and its preparation method and an application. The warm starting TaqDNA polymerase is prepared by an antibody modification method, and includes steps of (1), structuring and transformation of a recombination expression carrier; (2), inductive expression of recombinant bacteria; (3), preparation and purification of crude enzyme; (4), antibody modification. The warm starting TaqDNA polymerase is high in specificity, high in sensitivity, and convenient to operate; the warm starting TaqDNA polymerase can be applied to regular PCR amplification, real-time fluorogenic quantitative PCR, multiplex PCR, and digital PCR and TA cloning.

The invention relates to primers and TaqMan probes used for carrying out two-channel fluorescence PCR detection of trichomonas vaginalis viruses and candida albicans viruses. The primers and the TaqMan probes are trichomonas vaginalis virus primers and trichomonas vaginalis virus TaqMan probes designed based on a trichomonas vaginalis virus conserved area and candida albicans virus primers and candida albicans virus TaqMan probes designed based on a candida albicans virus conserved area. Simultaneously, the primers and the TaqMan probes are used for detecting whether trichomonas vaginalis virus RNA and candida albicans virus DNA exist in a sample to be detected. The invention further comprises a detection method employing the primers and the TaqMan probes and a kit containing the primers and the TaqMan probes.

The invention belongs to the field of analytical chemistry, and relates to a method for detecting bisphenol A based on a quantum dot-gold nanoparticle self-assembled superstructure. The method comprises the following steps: at first, respectively preparing a fluorescent quantum dot and a gold-nanopartcle, preparing the quantum dot-gold controlled nanoparticle self-assembled superstructure by modifying a DNA on the surface of a nano materia and taking a DNA molecule as a template. The quantum dot is enabled to be separated from an assembly by using the specific recognition of bisphenol A and an aptamer modified on the surface of the quantum dot; the concentration of bisphenol A is detected by the change of fluorescence intensity in a system, the detection line of bisphenol A is 1.64 pg/mL, and the linearity range is 1-500 pg/mL. Quick and highly sensitive detection of bisphenol A can be achieved specifically, and an effective method is provided for detecting traces of noxious substances in food packaging materials.

The invention provides a campylobacter jejuni fluorescence quantitative PCR detection method and a detection kit thereof. The method can rapidly and accurately detect the campylobacter jejuni and reduces the false positive. The kit comprises the following reagents: PCR MIX reaction liquid, 5*FQ PCR buffer, a primer mixture with the concentration of 10 mu M, a TaqMan probe mixture with the concentration of 10 mu M, and dNTPs and ddH2O with the concentration of 10 mM, wherein the primer mixture comprises a 16S rRNA upstream primer F, a 16S rRNA downstream primer R, and an upstream primer hipO-F and a downstream primer hipO-R; and the TaqMan probe mixture comprises: (1) 16S rRNA-P and hipO-P; (2) Taq enzyme; and (3) positive standard substances and negative standard substances. By introducing high-efficiency and strong-specificity amplification primers and fluorescent probes, the sensitivity and specificity of detecting the campylobacter jejuni are improved and missing detection and error detection are avoided.

The invention discloses a specific primer for diagnosing nosema antheraeae and an application of the primer. High-sensitivity specific detection for the nosema antheraeae is realized by an SYBR Green fluorescent quantitative PCR (polymerase chain reaction) method, the optimal reaction condition includes 95 DEG C and 30 seconds, circulation is performed for 40 times under the condition of 95 DEG C and 5 seconds and 62.5 DEG C and 30 seconds, and the melting curve condition includes 95 DEG C, 15 seconds, 60 DEG C, 60 seconds, 95 DEG C, 15 seconds, 60 DEG C and 15 seconds. Detection sensitivity is improved by two magnitude orders as compared with that of disclosed PCR technology, and lower detection limit includes that each reaction system contains 0.0046ng nosema genome DNA (deoxyribonucleic acid). Reliability of detection results is quantized by calculating primer amplification efficiency. The specific primer effectively solves the problems of low sensitivity, late detection period, easiness in leak detection, environmental pollution caused by conventional PCR nucleic acid coloring agents and the like, and has a wide application prospect in terms of antheraeae pebrine inspection and quarantine and cocoon quality improvement.

The invention relates to the technical field of biology, and concretely discloses application of echinococcus granulosa glutaredoxin-1 to preparation of an immunizing antigen and/or a kit for detecting echinococcosis granulose. When being used as an immunizing antigen, the echinococcus granulosa glutaredoxin-1 can be recognized by sheep positive blood serum naturally infecting the echinococcosis granulose. When being applied to indirect ELISA detection, the echinococcus granulosa glutaredoxin-1 has high specificity and sensitivity; the clinic detection coincidence rate is as high as 97.9 percent. The detection method built by using the echinococcus granulosa glutaredoxin-1 as the immunizing antigen has a good diagnosis effect, and can be used for the primary screening of the echinococcus granulosa of sheep in an infected area.

The invention relates to a reagent kit for identifying breast cancer states or breast precancerous lesion and application of the reagent kit. The reagent kit comprises primer pairs for detecting methylation levels of biological marker genes or corresponding segments in biological samples from subjects. The primer pairs are used for carrying out PCR (polymerase chain reaction) by the aid of the biological marker genes or the segments of the biological marker genes, the biological marker genes or the segments are used as templates, and the biological marker genes are treated by hydrosulfite; thebiological marker genes are selectively a type or a plurality of types of APC, BRCA1, CCND2, CST6, GP5, GSTP1, PITX2, RARB, RASSF1A and SOX17. The reagent kit and the application have the advantagesthat the methylation levels of the biological marker genes or the corresponding segments are jointly detected, the sensitivity and the specificity of breast precancerous lesion and breast cancer detection can be improved, accordingly, the correctness and the reliability of detection results can be guaranteed, and a novel quick, reliable and accurate way can be provided to predicting, diagnosing and evaluating breast cancer.

The invention discloses a colon cancer specific antigenic peptide, an expression vector and a preparation method thereof, a colon cancer specific autoantibody enzyme-linked immune detection kit comprising the expression vector and an operation method thereof. The preparation method comprises the following steps of: showing a colon cancer cDNA library onto the surface of a phage by utilizing the phage; screening an expression vector of the colon cancer specific antigenic peptide from the colon cancer cDNA library through affinity screening and serology analysis as a main component; and formingthe detection kit by the main component with a negative control, an elisa plate, a color developing agent, a stop solution, and the like. The expression vector of the colon cancer specific antigenic peptide effectively expresses the colon cancer specific antigenic peptide, is coated on the elisa plate and has the capacity of detecting autoantibodies generated by tumor simulation in the blood serum of a colon cancer patient so as to achieve the purposes of early detecting and screening colon cancer patients.

The invention relates to a prostate cancer early diagnosis or prognosis evaluation marker lncRNA malat1 and application thereof. The marker is an exosome-derived long-chain non-coding RNA (lncRNA) malat1, has a good indicating effect on the early diagnosis of a prostate cancer, can realize the non-invasive detection on the prostate cancer and has higher sensitivity and specificity when being compared with a conventional prostate cancer non-invasive detection method, thereby clinically avoiding the unnecessary puncture biopsy for prostate cancer suspected patients. In addition, the marker alsohas a good prediction effect on the prognosis of the prostate cancer and favorably guides the recurrence monitoring and the early clinical intervention of the patients.

The invention discloses an anti-H3N2 subtype canine influenza virus nucleoprotein monoclonal antibody hybridoma 2D10, and a monoclonal antibody. The above hybridoma cell strain is preserved in China Center for Type Culture Collection on August 12, 2016 with the preservation number of CCTCC NO:C2016155. The hybridoma cell strain obtained in the invention has high secretion output, and the monoclonal antibody prepared for an H3N2 subtype canine influenza virus NP has high specificity and sensitivity, can bind to a denaturated NP protein in ELISA and Western-blot detection, can bind a natural NP protein with a high-grade structure in immunofluorescence detection, can provide scientific bases for researches of the structure functions of the NP protein and development of various subtype in-vitro diagnosis reagents, and provides experiment data support for the feasibility of the monoclonal antibody used as a disease marker.

The invention provides a screening method and screening device of microsatellite sites pertinent to genome stability, and an application. The screening method comprises the steps of determining the state of microsatellite sites of respective tumor tissue and blood cells of a plurality of samples and the sample state of the samples by a first method; performing verification on the sample state of the samples by a second method; selecting the samples consistent in the sample state, determined by the first method and the second method, and dividing the samples consistent in the sample state, intoMSS first-class samples and MSI-H second-class samples; respectively screening out a first site set formed by a plurality of microsatellite sites highest in relevance with the first-class samples, and a second site set formed by a plurality of microsatellite sites highest in relevance with the second-class samples; and taking the intersection of the first site set and the second site set to obtain the microsatellite sites pertinent to the genome stability. The microsatellite sites screened by the method are high in detection accuracy and high in detection sensitivity and specificity.

The invention belongs to the technical field of biology and discloses a primer set for the detection of a koi herpesvirus Sph gene and the application of the primer set. The premier set for the detection of the koi herpesvirus Sph gene comprises a forward outer primer having the nucleotide sequence shown in SEQ ID No.1, a backward outer primer having the nucleotide sequence shown in SEQ ID No.2, a forward inner primer having the nucleotide sequence shown in SEQ ID No.3 and a backward inner primer having the nucleotide sequence shown in SEQ ID No.4. The primer set provided by the invention is high in specificity and sensitivity, and suitable for amplification of koi herpesvirus Sph gene through an LAMP (Loop-Mediated Isothermal Amplification) method, and can be used for preparation of a reagent for detection of the koi herpesvirus Sph gene.

The invention relates to an anti-glyphosate resistance protein 5-enolpyruvyl-shikimate-3-phosphate synthase (EPSPS) monoclonal antibody in commonly seen in transgenic crops and a preparation method thereof. The anti-CP4 EPSPS monoclonal antibody is a novel monoclonal antibody for detecting if a natural transgenic plant (such as soybean, corn, cotton or paddy rice) contains a CP4 EPSPS protein. A special-purpose antigen of the anti-CP4 EPSPS monoclonal antibody is obtained by recombination expression of a published CP4 EPSPS gene sequence in escherichia coli. The subtype of the anti-CP4 EPSPS monoclonal antibody is IgG1 and has affinity constant of 2.35*10<8>. The anti-CP4 EPSPS monoclonal antibody can identify a CP4 EPSPS protein in the transgenic plant and can be used for detection of a plant with the CP4 EPSPS gene. The invention also provides light chain and heavy chain variable-range coding sequences of the anti-CP4 EPSPS monoclonal antibody.

The invention discloses a method for measuring nitrofuran antibiotics in cosmetics. The method comprises the following steps: (1) pretreating a sample, namely adding the sample into a mixed solution of acetonitrile and methanol in a volume ratio of 70:30, uniformly mixing, performing ultrasonic extraction, centrifuging, airing supernatant till the supernatant is nearly dry, dissolving residues in a mixed solution of a 15mmol/L ammonium acetate solution and acetonitrile according to a volume ratio of 85:15, and sieving the solution through a micro-porous filter membrane with the size of 0.22 micron, thereby obtaining filtrate for later use; (2) performing high performance liquid chromatography separation on the filtrate, eluting a mobile phase by a gradient elution program, and quantifying according to an external standard method; and (3) confirming a detected positive sample by high performance liquid chromatography-mass spectrometry/mass spectrometry. According to the method, four nitrofuran antibiotics such as macrodantin, furazolidone, furaltadone and nitrofurazone are simultaneously detected. The method is accurate, reliable and high in sensitivity.

The invention discloses a detection kit of ochratoxin and a method thereof for detecting the ochratoxin, and belongs to the technical field of detection of harmful substances. The detection kit of ochratoxin disclosed by the invention comprises a nucleic acid aptamer of the ochratoxin, a cDNA which is not completely complementary to the nucleic acid aptamer, and two hairpin sequences H1 and H2, wherein the sequence of the nucleic acid aptamer of the ochratoxin is as shown in a sequence table SEQ ID NO. 1; the two hairpin sequences H1 and H2 can be self-assembled into a long chain; and the cDNAwhich is not completely complementary to the nucleic acid aptamer of the ochratoxin can trigger the self-assembly of the hairpin sequences H1 and H2. The kit provided by the invention has very high sensitivity and specificity, simple operation and rapid detection, the result is visible to naked eyes, the kit is suitable for promotion in all laboratories, and real-time onsite analysis of the ochratoxin can be realized without professional training.

The invention discloses an anti-p16 monoclonal antibody. A heavy chain variable region amino acid sequence of the anti-p16 monoclonal antibody is encoded by the DNA sequence shown in SEQ ID No.2. A light chain variable region amino acid sequence of the anti-p16 monoclonal antibody is encoded by the DNA sequence shown in SEQ ID No.3. The heavy chain variable region amino acid sequence of the anti-p16 monoclonal antibody is the amino acid sequence shown in SEQ ID No.4. The light chain variable region amino acid sequence of the anti-p16 monoclonal antibody is the amino acid sequence shown in SEQ ID No.5. The monoclonal antibody generated through hybridoma (30859-S17) secretion can recognize recombinant protein p16 molecules and lymphocytes expressing p16, and various kinds of tumor tissue of skin squamous-cell carcinoma, pancreatic cancer, melanoma, lymphoma, esophageal carcinoma, cervical cancer and the like of high-expression p16 protein can be detected.

A system and a method are described for applying a noise model for predicting the occurrence and a level of noise that is present in cfDNA read information. The significance model is trained for a plurality of stratifications of called variants using training data in the stratification. Stratifications may include a partition and a mutation type. The significance model predicts the likelihood of observing a read frequency for a called variant in view of two distributions of the significance model. The first distribution predicts a likelihood of noise occurrence in the sample. The second distribution predicts a likelihood of observing a magnitude of the read frequency for the called variant. The two distributions may further depend on a baseline noise level of blank samples. With these two distributions, the significance model, for a particular stratification, more accurately predicts the likelihood of a false positive for a called variant.

The invention discloses PCR primer groups and a probe used for detecting human immunodeficiency virus type I, a kit containing the PCR primer groups and the probe, and a detection method. The kit contains the high sensitivity primer groups and the probe. According to the detection method, multiple circulation is designed, and temperature is increased to a primer annealing temperature, and extremely high sensitivity and specificity are achieved. According to the minimum detection limit, in each time of detection, 2 copies HIV-1 DNA/ millions of cells can be realized, and sensitivity of the detection method is higher than that of other detection methods. A cell quantitative system is introduced into the detection method, so that quantification of HIV-1 DNA and cell number can be realized simultaneously in a same reaction. A quantitative standard substance is added into the kit so as to solve a problem of HIV DNA auantitative traceability in the prior art, wherein the quantitative standard substance is simple to prepare, is stable, and is reliable in traceability. The PCR primer groups, the probe, the kit, and the detection method can be used for HIV infection early detection, HIV infection screening of infants or high risk groups, HIV-1 virus repository detection, antiviral drug therapeutic effect evaluation, disease relapse control, and radical cure standard establishment; operation is simple; cost is low; clinical application prospect is promising; and it is beneficial for popularization and application.

The invention discloses application of EG-TPx and a kit for diagnosing echinococcosis granulosa of livestock. The EG-TPx can serve as an immunizing antigen and can be recognized by positive sheep serum with natural infection of echinococcosis granulosa. When applied to indirect ELISA (Enzyme-Linked Immuno Sorbent Assay), the EG-TPx has excellent immunogenicity and high specificity and sensitivity,the sensitivity is 92.6%, the specificity is 99.0%, and the cross reaction with other taeniasis positive serum is low. The ELISA detection method established by an immunizing antigen prepared from echinococcus granulosus thioredoxin peroxidase has an excellent diagnosis effect, and can be applied to preliminary screening of echinococcosis granulosa of cattle and sheep in epidemic areas.

The invention provides an isolated binding protein bound with NS1 protein and in an antigen binding structural domain. The binding protein comprises a specific heavy chain CDR and a light chain CDR and is capable of specifically recognizing and being bound with the NS1 protein and high in sensitivity and specificity, so that dengue virus detection is realized. In addition, the binding protein doesnot need to be produced from mouse peritoneal induced hybridoma cells, and antibody functional stability is improved while low production difficulty is realized.