559results about How to "Guaranteed specificity" patented technology

The invention discloses a human leukocyte antigen HLA-A and HLA-B gene full-length sequencing method and an HLA gene sequencing and typing method. The HLA-A and HLA-B gene full-length sequencing method comprises the following steps of: a, performing PCR amplification on about 4kb full-length sequences of HLA-A and HLA-B genes by using a pair of primers respectively; and b, cloning the amplification products to a pGEM-Tea sy vector, sequencing the full-length sequences by using ten walking sequencing primers in positive and negative directions respectively, and totally obtaining 38 allele 4.3kb full-length sequences of the HLA-A and 30 allele 3.7kb full-length sequences of the HLA-B. The HLA-A and HLA-B sequencing and typing method comprises the following steps of: performing PCR amplification on typing target areas of the HLA-A and HLA-B by using two pairs of primers respectively; and performing two directional sequencing on products by using fourteen sequencing primers respectively, wherein the HLA-DRB1 and HLA-DQB1 sequencing and typing method comprises the following steps of: amplifying sequences of second and third exons of DRB1 and DQB1 by adopting group specificity primers respectively; performing two directional sequencing on the second and third exons of the DRB1 by adopting eight group specificity primers and three sequencing primers; and performing two directional sequencing on the second and third exons of the DQB1 by adopting four sequencing primers respectively.

A method for preparing oligonucleotide chip for transgene product detection and use are disclosed. It includes: designing specific oligonucleotide probe and related primer by heterogeneric inserting gene, species internal standard gene, specific boundary sequence in transgene product gene set, fixing probe on glass slide to form transgene product detecting chip, amplifying DNA of plant to be tested by related primmer, marking by fluorescence, and hybridizing with chip. It can be use to acquire variable information.

The invention relates to a small RNA/DNA whitening product series, which mainly consists of a small RNA skin whitening lotion and a small RNA skin beauty cream. The small RNA skin whitening lotion mainly comprises efficient active ingredients for removing skin pigmentation and whitening skin, such as nano single-strand or double-strand oligonucleotides for specially and efficiency restraining genes (Asip, Mclr, Mitf, Dct, Silv, Oca2, Myo5a, Loc51151, Rab27a, Pomc, OA1,F2rl1, Kinesin, Slc24a5, Tyr and Tyrp1) relevant to melanin generation and a transdermal agent for stimulating oligonucleotides to enter skin melanocytes; the small RNA skin beauty cream comprises a plurality of highly active materials for maintaining pure and white skin and small RNA segments to prevent the sunshine from damaging the protective components of the skin and remove free radicals, and anti-aging and anti-inflammatory effective molecules. In addition, the invention further provides a formulation, a method and a procedure for preparing the series of whitening products.

The invention discloses a method and a kit for adopting a colloidal gold chromatographic technique for detecting mycoplasma pneumoniae nucleic acid and belongs to the technical field of medical biochemistry. According to the method, a colloidal gold grain is directly marked on a nucleic acid probe; the sequence for the marked nucleic acid probe is designed as a universal sequence; the nucleic acid probe also can be used for detecting other pathogens. During the design process for the kit provided by the invention, the introduced special probe A and special probe B have the functions of bridge molecular components and a gold marked probe and an MP (Mycoplasma Pneumoniae) nucleic acid amplified fragment are successively combined with each other in series by the two probes, so that the special detection for the MP nucleic acid fragment is realized. More than two probes can be designed in each set of probes; such a design is beneficial to the increasing of the sensitivity of the test strip; the advantages of the amplification technique for the depending nucleic acid sequence of MP and the colloidal gold marked detection for the products after amplification are integrated; the technical demand on the experimenter is low; no special instrument is required; the popularization of the MP nucleic acid detection in basic and faraway rural medical institutions is easily realized.

The invention belongs to the field of biomedicine examination, and particularly relates to a kit and a method for detecting mycobacterium tuberculosis infection and application. The invention discloses a novel mycobacterium tuberculosis detection reagent by screening specific T cell epitope of mycobacterium tuberculosis, wherein the reagent contains polypeptide or analog thereof represented by SEQ ID No.1-10. The method detects cell factors released from T cells by using single or more SEQ ID No.1-10 polypeptides to contact the T cells of mycobacterium tuberculosis infected individuals. The method can effectively detect active tuberculosis or latent tuberculosis infection, and is free from disturbance of Bacilli Calmette Guerin (BCG) inoculation vaccines. The invention also discloses a diagnostic kit and other application based on the polypeptide and the method. Compared with the gamma interferon release experiments in the prior art, the method can obviously improve the detection rate without reducing the specificity and has high clinical application value.

The invention discloses a hyper-blocking fluorescence quantitative PCR method with high sensitivity for detecting rare mutation. The invention adopts specific primers and a probe technology and can rapidly detect gene point mutation in a blood sample. The method has the advantage that (1) the conventional ARMS primers are modified by LNA or mismatched bases are introduced into 3' tail 2th site totail 3th site so that the sensitivity of the mutation detection is not influenced and the specificity of the single base mutation based on the mutation specific primers is guaranteed, (2) the modifiedblocker probe completely complementary to the wild type gene is introduced so that the wild type background interference is further reduced with keeping the efficiency of the mutant amplification, (3) based on the high specificity modified blocker probe, the ARMS primers can detect different base mutation forms at the same site at the same time so that multiple sites in the same gene can be detected by only a few systems, (4) sensitivity is high and (5) a detection rate is fast.

The invention discloses a method for detecting the specific antibody of classical swine fever virus and a special ELISA kit thereof. The kit includes a classical swine fever virus antigen and an enzyme-labeled classical swine fever virus single-epitope specific antibody; the said swine fever virus antigen is a polypeptide containing one or more than one amino acid residue sequence described in sequence 1. The detection reagent of the classical swine fever virus specific antibody of the present invention can carry out effective detection to the classical swine fever virus specific antibody by solid-phase antigen competition ELISA (blocking method); The B cell epitope ensures the differential diagnosis, and the high degree of conservation of the epitope among various strains ensures the specificity of detection.

The invention discloses variant porcine reproductive and respiratory syndrome virus (PRRSV) TaqMan fluorescence quantitative RT-PCR detecting kit and application thereof. A primer and a TaqMan probe are designed and synthesized by referring to an NSP2 fragment gene sequence of the variant PRRSV and common PRRSV of a GenBank. By optimizing the reaction condition and constructing a standard plasmid product, a method for diagnosing the variant PRRSV by TaqMan fluorescence quantitative RT-PCR is established. A result indicates that the method has the advantages of strong specificity, high sensitivity, and the like and can detect the standard plasmid product with 264 copy numbers, and the virus quantity of 0.5623TICD50 is 10 times more sensitive than RT-PCR. By detecting 22 disease samples, 8 disease samples are positive, and the positive rate is 36.4 percent. Because the method has the advantages of quantification, high speed, accuracy, sensitivity, and the like, the invention is suitable for the diagnosis on the swinery infected variant PRRSV in the early stage, the medium stage and the later stage and plays an important role in effectively diagnosing, preventing and treating the highly pathogenic PRRSV.

The invention discloses an on-site detection immuno-chip, comprising a chip carrier and an agarose gel layer which is paved on the chip carrier, wherein, the agarose gel layer is fixed with a plurality of antibody microarrays of 4*4, and chip-dedicated fences or Super PAP Pen scribings are used for separating the microarrays from each other; the double antibody sandwich principle is adopted to detect the antigen to carry out the dot matrix of corresponding capture antibody, positive control and negative control on the solid phase carrier simultaneously; the antibody protein is connected with the solid phase carrier through the covalent bond and physical adsorption; the sample liquid to be detected and the chip are directly incubated; the antigen to be detected in the samples combine with the corresponding antibody which is fixed on the chip; a specific monoclonal antibody probe marked by horse radish peroxidase is added and the macroscopic detection results can be obtained after the coloration of substrates. The invention can detect viruses in multiple aquatic animal samples and the results are macroscopic, thus being applicable to rapid and accurate detection of viruses of aquatic animals in breeding production.

The invention discloses an immunochromatographic test strip, and a production method and an application thereof. The test strip is characterized in that a labeling compound carries an inert protein connected with a special label and used for quality control, and a quality control line contains a monoclonal antibody specifically binding to the special label carried by the inert protein. The test strip has an independent quality control system, so the obtained test result has a high specificity and an anti-interference property, non-specific reactions brought by the self-bearing antibodies in human blood samples are effectively avoided, and the effectiveness of the test strip is effectively indicated.

The invention relates to the field of medicine synthesis, and discloses a method for synthesizing Etelcalcetide. The method comprises the following steps: performing liquid-phase synthesis so as to obtain straight-chain heptapeptide which is subjected to acetylization at an N end and is subjected to amidation at a C end on the amino acid sequence of SEQ ID NO:1; performing NCS (Chlorosuccinimide) chlorination on L-Cys, and substituting chlorine on the sulfydryl, and generating L-Cys (SCl); performing coupling reaction on the straight-chain heptapeptide and L-Cys (SCl) to generate disulfide, thereby obtaining Etelcalcetide. As the Etelcalcetide is synthesized by using a whole liquid-phase method, relatively small amounts of reagents and solvents are used when being compared with those of a solid-phase method, the synthesis is green and environmental-friendly, no expensive resin is used, and thus the cost is lowered. Meanwhile, as NCS which is cheap and easy to obtain and cysteine are adopted to form an active intermediate to construct the disulfide bond, the defects that a conventional method is low in conversion rate and low in purity are avoided, and large-scale production of the Etelcalcetide can be facilitated.

The invention provides a reagent for detecting brucella. The reagent comprises an upstream primer, a downstream primer, a fluorescence probe and a quenching probe; the gene sequence of the upstream primer is 5'-caagggcaaggtggaagatt-3'; the gene sequence of the downstream primer is 5'-ctgcgaccgatttgatgttt-3'; the gene sequence of the fluorescence probe is 5'-fam-atcgtttccgggtaaagcgtcgcca-P-3'; and the gene sequence of the quenching probe is 5'-cgctttacccggaaacga-Dabcyl-3'. The invention also provides a complex probe fluorescence quantitative polymerase chain reaction (PCR) brucella detection method using the reagent. The method is simple and convenient in operation, efficient, quick and specific; the detection time of the brucella is greatly shortened; the quantitative detection of a sample can be completed in about 2 hours; and the reagent and the method have significance for early diagnosis of brucella disease.

The invention provides a fast multiplex PCR identification diagnosis detection method of invasive fungus infection based on cfDNA (cell free DNA). The method can be used for identifying the invasive infection caused by clinic common and high-incidence Candida albicans, Candida tropicalis, Candida parapsilosis, Candida krusei, Candida glabrata and Aspergillus fumigtus. An amplification primer is designed according to characteristic genome segments of each fungus category and species; a detection fluorescence probe of a strain can be distinguished according to the amplification segment design; the real time PCR can be performed on a sample to be tested; high sensitivity of nested PCR and high specificity and multi-target performance advantages of multiple fluorescent hybrid probe PCR are integrated for identifying the fungus strain. The invention also provides a PCR diagnosis kit for the invasive fungus infection and application thereof.

The invention discloses a method and kit for realizing multiple detection of nucleic acid through a colloidal gold chromatographic technology, and belongs to the field of medical biotechnology. The invention provides a nucleic acid detection test strip, a universal probe is marked with colloidal gold particles to be fixed on a glass cellulose film, the universal probe is designed into a universal sequence, an NC film on the test strip have one or more detection lines and a quality control line, each detection line is coated with a section of nucleic acid sequence which can be in specific hybridization combination with a corresponding specific probe B series; the quality control line is coated with a section of nucleic acid sequence which can in specific hybridization combination with a specific probe A series; the specific probe A series is in complementary pairing hybridization with the universal probe; the probe marked with gold and a nucleic acid amplified fragment are combined in series by virtue of the specific probe A series and the specific probe B series, so that the specific detection of the nucleic acid fragment can be realized. The method has the advantages that the technical requirements of experimenters are low, the required detection time is short, special instruments are not required, and the method is easy to popularize towards grass-roots and remote countryside medical institutions.

The invention provides a Taqman fluorescent hydrolysis probe and a method for quantitatively detecting the methylation level of MGMT (O6-methylguanine-DNA-methyltransferase) by using the Taqman hydrolysis probe. The fundamental sequence of the Taqman fluorescent hydrolysis probe is GATTTGGTGAGTGTUTGGG, and the complementary region of the fundamental sequence and a genomic DNA (deoxyribonucleic acid) sequence extends upstream no more than 5nt and extends downstream no more than 5nt. According to the invention, the specificity of PCR (polymerase chain reaction) is doubly guaranteed; the percent of a methylated DNA template in a mixed sample can be quantitatively calculated; and the method has all the advantages (such as speediness, sensitiveness, no pollution, and the like) of the real-time quantitative PCR. The application of detection results of the invention can provide clinical guide for medicaments for carrying out chemotherapy on malignant tumors such as cerebral gliomas and the like; and the method has the advantages of high specificity, high sensitivity, accurate quantification, and the like.

The invention discloses three-party authentication method and device as well as an intelligent card supporting two-way authentication. The three-party authentication method comprises the following steps: authentication is performed between a terminal and the intelligent card; after the authentication between the terminal and the intelligent card is passed, the terminal reports a binding relationship between the terminal and the intelligent card to a management platform, and requests authentication of the binding relationship to the management platform; the management platform performs authentication of the binding relationship between the terminal and the intelligent card, if the authentication of the binding relationship is passed, the three-party authentication is judged to be passed, or else, the three-party authentication is judged not to be passed. By adopting the three-party authentication method and device as well as the intelligent card supporting two-way authentication, the safety of the terminal and the intelligent card are both ensured, and meanwhile, the binding relationship between the terminal and the intelligent card can be dynamically authenticated. The management platform side has a control and management right for the terminal and card equipment so that an operator can conveniently develop own business, and the specificity and safety of the terminal and the intelligent card during development of business of Internet of things are ensured.

The invention discloses a method for simultaneously detecting multiple anti-tumor drugs in a blood sample. A pretreated sample to be detected is detected by adopting ultra-high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The pretreatment process comprises the following steps: adding serum into a mixed solution of methanol and acetonitrile, oscillating and centrifuging,taking out the centrifuged supernatant, drying, dissolving the dried powder into a methanol aqueous solution, and filtering to obtain a sample to be detected. The method can be used for simultaneously detecting 13 kinds of anti-tumor drugs such as methotrexate, 5-fluorouracil, apatinib, busulfan, carboplatin, cyclophosphamide, docetaxel, gemcitabine, imatinib, illinotecan, lenalidomide, oxaliplatin, paclitaxel and the like in blood.

The invention discloses a multi-RCA (rolling circle amplification) method based on split padlock probes. Each novel split padlock probe is about 90bp in length and comprises four parts, namely a detection arm, a universal primer area, an HhaI endonuclease site and a tag sequence area, wherein an amplification system is composed of a connection system and an RCA system. The concrete detection method comprises the steps of: firstly, carrying out coupled reaction, mixing a target sequence DNA (deoxyribonucleic acid) segment and four split padlock probes of which the final concentration is 1mol/L, hybridizing for 15 minutes after boiling and degenerating, adding T4DNA ligase and T4DNA ligase buffer solution, supplying 10 muL of reaction system; carrying out exonuclease I and exonuclease III exterior contact for 45 minutes at 37 DEG C, preparing an annular template and then carrying out RCA reaction; mixing, boiling and degenerating 10 muL of connection product and universal primer, respectively adding dNTP, phi29DNA polymerase, HhaI restriction enzyme and buffer solution to form 20 muL of reaction system, stewing for 60 minutes at 37 DEG C; finally detecting single-chain DNA product of RCA with a specific tag sequence, thus obtaining a corresponding test conclusion according to the result.

The invention discloses a forward and reverse ABO typing and RhD blood type detecting card comprising a detecting card with six micro-columns, wherein the first three micro-columns in the six micro-columns are respectively filled with equivalent A-resistant gel, B-resistant gel and D-resistant gel, and the left three micro-columns are filled with equivalent blank gel. The forward and reverse ABO typing and RhD blood type detecting card is prepared by using the following method comprising the steps of swelling dextran gels; preparing all antibody diluents; preparing all antibody working solutions; washing the gels; and subpackaging to form the RhD blood type detecting card. According to the forward and reverse ABO typing and RhD blood type detecting card, crosslinked dextran gels with different sizes and sieving ranges are mixed and swelled by using purified water, so that the specificity is effectively ensured, meanwhile, the forward and reverse ABO typing and RhD blood type detecting card has very high sensitivity, and the ion strength of a system is effectively ensured.

The invention relates to a method for detecting, genotyping and quantifying human intestinal flora, consisting of universal primer PCR amplification and specific probe LDR. In the PCR amplification, the 16sr DNA fragment of selected bacteria genus is amplified by the designed universal primer synchronously; the specific LDR probe of each bacteria genus is utilized to achieve the goal of genotyping through ligation reaction; and LDR signal is detected by ABI Prism 377 type sequencer, and the detected fluorescence value is taken as the quantifying basis. Aiming at the universal primer of intestinal microflora, the detection method of the invention effectively amplifies all target intestinal microflora 16sr DNA sequence, thereby ensuring that each target sequence and the template of low copy therein can be detected, further ensuring the specificity of the specific probe of each bacteria genus, completely identifying each target sequence and realizing multiplex genotyping quantification.

The invention relates to primers and a method for detecting SNP gene typing of AhFAD2B genes of peanuts in high throughput. The primers comprise the specific primer of which the nucleotide sequence is shown as SEQ ID No.1 and SEQ ID No.2 and a universal prime of which the nucleotide sequence is shown as SEQ ID No.3. According to the primers and method, the allelic variation condition of the AhFAD2B genes can be detected and identified in high throughput, the maximum sample quantity of one-time detection can reach 1536, the detection cost is low, rapidness and accuracy are achieved, and the efficiency and accuracy of peanut breeding selection are greatly improved.

The invention relates to the fields of biotechnologies and medicines, particularly discloses a detection primer for EML4-ALK (anaplastic lymphoma kinase) fused gene mutation, a probe and a detection kit, and aims to detect the EML4-ALK fused gene mutation. The detection primer comprises sequences as shown in SEQ ID NO: 1 to SEQ ID NO: 14. The sequence of the probe is as shown in SEQ ID NO: 15, of which the end 5' is modified with FAM or VIC (vasoactive intestinal contractor), and the end 3' is modified with NFQ-MGB (non-fluorescent quencher-myohemoglobin). The kit comprises the detection primer, the probe and a quality control system. The detection primer can specifically identify 13 EML4-ALK fusion mutation types, is high in sensitivity and can detect 5-copied mutations; the whole reverse transcription PCR (polymerase chain reaction) and fluorescence PCR detection process can be completed only by 80 minutes.