111 results about "Molecular diagnostic techniques" patented technology

The invention relates to the technical field of clinical molecular diagnosis, in particular to a superficial modification based 5hmC (5-hydroxymethylcytosine) polymolecular marker and a CRC (colorectal cancer) early diagnosis model. With the adoption of a 5-hmC high throughput sequencing technology, 5-hmC expression quantities of peripheral blood plasma DNA whole genomes of a CRC patient and a healthy person are checked, 5-hmC differential expression of genetic loci of the whole genomes of the two groups of samples are compared, the genetic locus of the part with the most obvious differentialexpression is screened as the gene marker of the diagnosis model, and then the CRC early diagnosis model is constructed. Verification proves that the diagnosis sensitivity and specificity are both higher, and compared with a stool occult blood detection kit and a spetin9 kit in the market currently, sensitivity and specificity are improved by about 20% and 8% respectively. The diagnosis method hasthe advantages of being noninvasive in detection and convenient, can be applied to the first-line clinic and is used for early screening of CRC.

The invention discloses a nucleotide sequence and a kit for detecting a human epidermal growth factor receptor-2 (HER-2), which belong to the technical field of molecular diagnosis. A primer and a probe for detecting the HER-2 provided by the invention can be used for specifically amplifying and detecting amplification of an HER-2 gene. The invention also discloses the detection kit for detectingthe HER-2 gene based on a microdroplet type PCR (Polymerase Chain Reaction) system, so that the amplification of the HER-2 gene can be quickly, sensitively and accurately detected. The nucleotide sequence and the kit provided by the invention have the advantages of simplicity and convenience in operation, high specificity, high sensibility, low cost, high throughput and the like, can be used for quickly detecting clinic HER-2 gene amplification, and provide references for diagnosis and treatment of the clinic human epidermal growth factor receptor-2.

The invention discloses a multiple touchdown PCR (polymerase chain reaction) detection kit capable of quickly detecting acinetobacter baumannii in lower respiratory tract specimens, belonging to the technical field of molecular diagnosis of infectious diseases of lower respiratory tracts and aiming to solve quick diagnosis of acinetobacter baumannii in lower respiratory tract specimens. The kit comprises 10*PCR buffer solution, MgCl2, dNTP, TaqDNA polymerase, BSA (bovine serum albumin), acinetobacter baumannii positive control DNA and three pairs of acinetobacter baumannii primers. The invention discloses the multiple touchdown PCR detection kit and a detection method capable of detecting acinetobacter baumannii and adopts agarose gel electrophoresis to detect the PCR products. The multiple touchdown PCR detection kit and the detection method have the advantages of quickness, simpleness and convenience, specificity and sensitivity and can be used for quick diagnosis and epidemiological survey of acinetobacter baumannii infection.

The invention discloses a nucleotide sequence for detecting BRAF gene V600E mutation and application thereof and belongs to the technical field of molecular diagnosis. A primer and a probe which are used for detecting BRAF can be specifically amplified and are used for detecting the BRAF V600E gene mutation. The invention further discloses a detection kit for detecting the BRAF V600E gene mutationbased on a micro-droplet type digital PCR (Polymerase Chain Reaction) system and the mutation of a BRAF gene can be rapidly, sensitively and accurately detected. The nucleotide sequence has the advantages of convenient operation, strong specificity, high sensitivity, low cost, high flux and the like, can be used for rapidly detecting the clinical BRAF V600E gene mutation and provides reference for diagnosis and treatment of the BRAF V600E mutation.

The invention provides a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) reagent for detecting 2019-nCov and application of the CRISPR reagent in preparation of a kit for detectingthe 2019-nCov. The CRISPR reagent comprises an RPA (Recombinase Polymerase Amplification) primer pair and corresponding crRNA. Based on the CRISPR molecular diagnosis technology, through coupling of RPA-CRISPR, two-stage amplification of 'sequence amplification' (RPA completion) and 'enzymatic cascade' (Cas enzyme completion) can be realized, so that 2019-nCov is detected with high sensitivity andhigh specificity under the condition of a simple instrument (metal bath).

The invention belongs to the technical field of molecular diagnosis, and particularly relates to IncRNA application, a kit and medicine. According to the application of IncRNA for preparing the kit for diagnosis and / or prognosis evaluation of NSCLC. The nucleotide sequence of the IncRNA is shown in SEQ ID No.1. The IncRNA is highly expressed in plasma before the NSCLC patient surgery, after the surgery, the expression level in the plasma is remarkably lowered, the expression levels in the NSCLC cancer tissue, the NSCLC patient plasma and the NSCLC cells are all remarkably higher than those ofpara-carcinoma tissue, healthy human plasma and normal bronchus epithelial cells. The study result suggests that the IncRNA has the remarkable capability for inhibiting the NSCLC cell tumor activity,and the IncRNA is newly found for tumor inhibition.

The invention provides application of CD177 in preparation of a product for diagnosing biliary atresia, and relates to the technical field of molecular diagnosis. According to the invention, by detecting a large number of biliary atresia child patient samples and combining animal experiments, it is proved that the number of CD177+ neutrophile granulocytes in biliary atresia peripheral blood and liver tissue can serve as a powerful index for evaluating the liver injury degree at first; and therefore, detection of CD177 genes, CD177 protein or the CD177+ neutrophile granulocytes can serve as a more convenient, direct and accurate method for clinically evaluating the illness state of biliary atresia child patients. The invention also proves that CD177 has a pro-inflammatory effect in biliaryatresia; and the inhibitor of CD177 can be used for research and development of drugs for treating biliary atresia.

The invention relates to a gene chip, an amplification reagent and a kit for detecting alpha-thalassemia, and belongs to a molecular diagnosis technology, wherein 3 non-deletion (alpha<CS>alpha, alpha<QS>alpha and alpha<WS>alpha) alpha-thalassemia and 4 deletion (--<SEA>, --<THAI>, -alpha<4.2> and -alpha<3.7>) alpha-thalassemia can be rapidly and simultaneously detected by combining direct PCR andreverse dot blot (RDB).

The invention provides a method for detecting genes of lung cancer and colorectal cancer based on an NGS method. The method comprises the following steps: carrying out sample collection, extracting DNA in a sample, carrying out molecular label library building, carrying out hybridizing with Panel, carrying out NGS sequencing, carrying out analysis aiming at data obtained after sequencing, and judging the gene type of the sample, wherein the Panel comprises SEQ ID No 1-SEQ ID No 618. The detection method provided by the invention belongs to a molecular diagnosis technology for realizing accurate treatment on patients suffering from lung cancer and colorectal cancer, liquid biopsy is supported, and the sensitivity achieves 0.1%.

The invention relates to a porcine circovirus type 2 rapid typing detection kit which belongs to the field of biological detection and is a kit based on a SMART detection system method. By the adoption of the kit, the technical problem that a porcine circovirus type 2 (PCV2a, 2b, 2d, etc.) typing detection method requires a complex operational program, high experiment skill, long experimental period and the like is solved. A rapid and accurate molecular diagnosis technical product can be provided for porcine circovirus type 2 typing. The kit provided by the invention can be used in system diagnosis of porcine circovirus type 2 rapid typing and used to guide prevention and treatment of the disease caused by porcine circovirus type 2. And mutation detection of each type sequence of the virus can be realized, and the kit is beneficial to molecular epidemiological investigation of the porcine circovirus, meat product quarantine inspection, viral surveillance of forage and water quality in a pig farm and the like.

The invention discloses polygene detection primers for predicting a risk of a distant recurrence of early breast cancer and belongs to the technical field of molecular diagnosis. The polygenes comprise BUB1B, BLM, TPX2, DTX2, PIM1, OBSL1, YWHAB, CLCA2, SF3B5, ESR1 and ERBB2. Primer sequences are shown as SEQ ID NO.1-22. The primers have good specificity, all amplicons generated by a PCR amplification are short, and detection sensitivity of RNA extracted from an FFPE sample can be enhanced. The invention further discloses a method for predicting a risk of a distant recurrence of early breast cancer. The relative expression quantity of the polygenes in the FFPE sample is quantified through SEQ ID NO.1-28, a statistical method of a distant recurrence genetic model is established, the risk of the distant recurrence of the early breast cancer can be predicted, patients with low-risk and high-risk breast cancer are divided, and guidance is provided for treatment of postoperative breast cancer.

The invention discloses a breast cancer tumor related tumor marker, an application and a kit, and relates to the technical field of tumor molecular diagnosis. The inventor, by research, finds that plant homeodomain zinc finger protein 20 (PHF20) is highly expressed in the breast cancer. And after an shRNA gene silencing method is used for down-regulating the expression level of the PHF20, the PHF20 has the obvious inhibition effect on tumor formation of breast cancer cells. The result shows that the PHF20 can be used as a drug regulation target in addition to the breast cancer marker. The invention provides a novel biomarker for breast cancer treatment and diagnosis. The PHF20 serving as the tumor marker of the breast cancer can be used for preparing the kit for breast cancer auxiliary diagnosis, curative effect prediction and prognosis judgment, and has the important clinical application value.

The invention discloses a nucleic acid analysis cassette and nucleic acid analysis equipment. The nucleic acid analysis cassette comprises a cassette body and a magnetic bead transferring component which is movably arranged on the cassette body along a set direction, wherein the cassette body is provided with at least two liquid storage chambers, sample application passages, exhaust passages and adetection structure, the at least two liquid storage chambers are distributed sequentially along a set direction; and the magnetic bead transferring component is provided with magnetic bead enrichingstructures, the liquid storage chambers, the sample application passages, the exhaust passages and the detection structure are all located at the same side of the magnetic bead transferring component, a surface where the liquid storage chambers are located is in tight connection with the magnetic bead transferring component, and the magnetic bead transferring component can enable the magnetic bead enriching structures to communicate with each liquid storage chamber sequentially. According to the nucleic acid analysis cassette, magnetic-bead-method-based nucleic acid extraction and amplified detection can be fully integrated, the compactness of the nucleic acid analysis cassette is improved, the size of the nucleic acid analysis cassette is reduced, and the popularization of molecular diagnosis technologies is promoted.

The invention discloses a gene composition, a kit and a method used for detecting pulmonary adenocarcinoma KRAS mutation, and belongs to the technical field of molecular diagnosis. The gene composition consists of the following 25 genes, including ELN, PODN, LAMC3, COL5A3, CRYAB, SOX18, AKT3, FAM101B, TIE1, SH3PXD2A, EXOC3L2, NOTCH1, NDST1, MMRN2, SH3RF3, ZMIZ1, RNF122, FLT4, F10, TLN1, SOX17, PIP5K1C, TMEM255B, WSCD1 and PCDHB3. In the invention, through the above gene composition, a Score scoring model is constructed for detecting the pulmonary adenocarcinoma KRAS mutation, and therefore, the gene composition has the advantages of high accuracy and good sensitivity and specificity, is convenient in operation, can carry out quantitative analysis and has a wide application prospect.

The invention discloses a eukaryotic expression method of a fish viral hemorrhagic septicemia virus G gene. The method includes the following steps of (1) gene synthesis of a main antigenic domain optimized by codons, (2) construction of a reconstructed baculovirus transfer vector, (3) construction of shuttle plasmids and (4) analysis and identification of reconstructed Bacmid transfection and expression products. By means of the method, efficient expression of the G protein antigenic domain in a Bac-to-Bac baculovirus system; the activity of the expression protein is primarily identified and analyzed through Western blotting, it is proved that the expression protein has immunogenicity, and a foundation is provided for further epitope vaccine development and VHSV molecular diagnosis technology establishment.

The invention relates to the technical field of molecular diagnosis, in particular to a composition for detecting mutation of lung adenocarcinoma cell cycle progress pathway related genes (CDKN2A, CCND1, CCNE1, CDK4 and RB1) and application of the composition, and the composition is composed of reagents for detecting the expression quantity of the related genes. RNA sequencing, LASSO and binary classification Logistic regression are applied to screen genes related to the cell cycle progress pathway, a score Score is constructed, a cut-off value corresponding to the Score in the lung adenocarcinoma is obtained through ROC analysis, and the method can be used for predicting mutation of the lung adenocarcinoma cell cycle progress pathway related genes. The specific gene mutation of the lung adenocarcinoma is predicted by using the expression quantity of the gene composition, and verification shows that the prediction method has the advantages of high accuracy and good specificity, and has a good application prospect.

The invention discloses a device used for rapid visualized detection of constant temperature nucleic acid amplification in the technical field of molecular diagnosis. According to the method, constant temperature nucleic acid amplification is adopted to amplify a target nucleic acid, a lateral flow chromatography process is adopted to perform visualized detection, and a whole system is sealed in a transparent box having a size of 9 cm*3.4 cm. Through a special structure design of the device, integration of nucleic acid amplification, sample delivery, development and result determination under a sealed condition is achieved, thus avoiding an uncovering need in detection, avoiding a false positive problem caused by nucleic acid contamination and overcoming the greatest barrier in nucleic acid visualized detection application. The device can achieve high-sensitivity nucleic acid visualized detection on the premise of no generation of nucleic acid contamination, and plays an important role in the fields of disease diagnosis, epidemic disease monitoring, prenatal screening, food safety detection, and the like.

According to the integrated nucleic acid detection chip and method based on the CRISPR technology, an isothermal amplification technology and a CRISPR molecular diagnosis technology can be carried out in one device at the same time, the problem that the two technologies are incompatible is solved through integrated detection, and through simple manual operation, the detection efficiency is greatly improved. A user can realize high-sensitivity nucleic acid detection, so that the support of large-scale instruments and equipment is avoided, and the use cost is greatly reduced. The device is convenient to carry, provides a good platform for separation of a CRISPR molecular diagnosis technology from a central laboratory, is convenient to operate, accelerates the nucleic acid detection speed, saves a part of medical cost, and solves the problems of slow detection result, high cost and incapability of realizing detection integration in the prior art.

The invention belongs to the technical field of molar-diagnosis and solves the problems that in the prior art, deterioration and metastasis of tumor cells are not known clearly and diagnosis evidencecannot be supplied to tumor metastasis and prognosis diagnosis. The invention provides an application of an AMP-activated protein kinase (AMPK) as a biomarker of tumor metastasis and prognosis diagnosis, and an application of AMPK[alpha]1 as a biomarker for the tumor metastasis and prognosis diagnosis. In the invention, an experiment proves that cancer signals (PI3K, Ras and Her2) inhibiting protein expression of the AMPK[alpha]1 in malignant tumors are matched with clinical data very well, thus proving that the significant low expression of the AMPK[alpha]1 in malignant tumors is a key factorof the metastasis of malignant tumors; with the gene expression level of the AMPK[alpha]1 as an important index for the diagnosis and prognosis of tumor metastasis, the AMPK[alpha]1 can be applied asa biomarker to the tumor metastasis and prognosis diagnosis.

The invention belongs to the technical field of animal pathogen molecule diagnosis, and particularly relates to a type A seneca virus detection primer, a kit, a virus detection method and application.The following primers including an SVA forward primer being SEQ ID NO.1:5'-AGAATTTGGAAGCCATGC-3' and an SVA reverse primer SEQ ID NO.2:5'-GAAACAGAYTGCAGCTTCTC-3' are comprised, wherein the Y represents a degenerate base group, and is a base group T or C. The high sensitivity and specificity are ensured; meanwhile, the detection accuracy is improved; the false positive and false negative probability is reduced.

The invention discloses a nucleotide sequence and a kit for detecting hepatitis C viruses (HCV), and belongs to the technical field of molecular diagnosis. By a primer and a probe for detecting the HCV, virus RNA of seven HCV genotypes can be specifically amplified and detected. By the kit for detecting the hepatitis C viruses, accurate copy numbers of various subtypes of the HCV can be detected quickly, sensitively and accurately. The nucleotide sequence and the kit have the advantages of simplicity and convenience in operation, high specificity, high sensitivity, low cost, high throughput and the like, can be applied to clinical quick hyper-sensitive detection of HCV virus RNA, and provide reference to clinical diagnosis and treatment of hepatitis C.

The invention belongs to the technical field of molecular diagnosis, and particularly relates to exosome miRNA serving as a molecular marker for diagnosing type 2 diabetes mellitus complicated with coronary heart disease and application of the exosome miRNA. The exosome miRNA is any one or a molecular marker combination of more than two of miR-15a-3p, miR-18a-5p, miR-19a-3p, miR-133a-3p, miR-155-5p and miR-210-3p, and the specific nucleic acid sequence of the exosome miRNA is shown as SEQ NO. 4, SEQ NO. 8, SEQ NO. 11, SEQ NO. 3, SEQ NO. 2 and SEQ NO. 1. Based on a total inventive concept, theinvention also comprises application of the exosome miRNA in preparation of a kit, a microarray or a biochip for diagnosing type 2 diabetes mellitus complicated with coronary heart disease. The invention proves that the six plasma exosome miRNAs can be used as molecular diagnosis markers of the type 2 diabetes mellitus complicated with coronary heart disease, and can be prepared into a kit, a microarray or a biochip for diagnosing the type 2 diabetes mellitus complicated with coronary heart disease, so that the type 2 diabetes mellitus complicated with coronary heart disease can be quickly diagnosed and judged by detecting the expression quantity of the plasma exosome miRNAs of a subject.

The invention provides a product for simultaneously detecting the polymorphism of MTHFR and MTRR genes, as well as a method and application thereof, and relates to the technical field of molecular diagnosis. The primer composition for simultaneously detecting the polymorphism of MTHFR and MTRR genes comprises a primer group for detecting rs 1801133 site, rs1801131 site and rs1801394 site, can be used for simultaneously and comprehensively detecting the polymorphic site of different types of genes with comprehensive detection with strong specificity and high sensitivity, can be used for accurately distinguish three genotypes of each site respectively, and can be used for detecting the lowest concentration of 1ng / mu l. The detection result can be used in clinical directed administration. Furthermore, locked nucleic acid is introduced into a probe for modifying, the probe with enough short length and excellent parting effect can be screened, each parameter of the probe accords with the designing principle, the accuracy of the detection result can be effectively improved, and an optimized parting effect is achieved.

The invention belongs to the technical field of molecular diagnosis, and particularly relates to a method and detection kit for detecting ERBB2 gene amplification based on a digital PCR technology. The kit comprises primer and probe premix liquid, wherein the premix liquid comprises upstream and downstream primers for detecting an ERBB2 gene, a fluorescent probe for detecting the ERBB2 gene, upstream and downstream primers for detecting a human ATCB reference gene, and a fluorescent probe for detecting the human ATCB reference gene. The method comprises the following steps of S1, providing a detection sample of a testee, wherein the detection sample is a plasma sample; S2, extracting ctDNA of a to-be-detected sample from the plasma sample in the step S1; S3, performing an amplification reaction on the sample extracted in the step S2 by using a digital PCR platform; and S4, analyzing and interpreting an amplification result. The ERBB2 gene amplification detection method provided by theinvention is based on a micro-droplet digital PCR detection system, reduces the interference of manual operation, has the advantages of stable result, high accuracy and good data amplification effect,and can be more conveniently used for auxiliary diagnosis and clinical treatment guidance.

The invention relates to a kit and a method for distinguishing more HPV (human papilloma virus) subtypes through a small number of detection reaction tubes by combining sequence combination with a nano-gold probe detection method. By a specific sequence combination, 21 typed human papilloma virus subtypes are distinguished by 6 detection tubes. By combining a mathematic permutation and combination algorithm with a molecular diagnostic technique, more human papilloma virus subtypes are distinguished by the smallest number of reaction tubes without increasing detection reaction flux, and detection operation difficulty and detection cost are reduced.

The invention relates to a chip, an amplification reagent and a kit for directly and simultaneously detecting alpha-thalassemia and beta-thalassemia mutation sites, and belongs to a molecular diagnosis technology. According to the present invention, based on direct multiplex PCR, Gap-PCR and reverse dot blot combined detection principle, the corresponding amplification primers and the corresponding probes are designed according to the mutation or deletion sites of each genotype, the primer is labeled with biotin, the probe is labeled with amino, a gene chip is used as substrate, the probe is immobilized on the DNA chip, the PCR product amplified by the specific primer is hybridized with the probe, and the diagnosis of thalassemia is performed by interpreting a signal coloring box.

The invention discloses lysate and kit for carrying out a PCR on a blood and oral cavity swab, and belongs to the technical field of molecular diagnosis. The kit comprises the lysate. The lysate is prepared from 60-70 mM of Tris-Hcl, 16-17 mM of ammonium sulfate, 6-7 mM of Mgcl2 6H2O, 6-7 micron M of EDTA, 1.5-2.5 micron M of SDS and 30-55 micrograms / ml of protease K, wherein the pH value of Tris-Hcl is 8.2-9. The blood and oral cavity swab treated with the lysate can be directly subjected to the PCR, and an existing detection technology is greatly optimized.

The invention relates to the technical field of molecular diagnosis, and discloses a hypersensitive microRNA electrochemical detection method of an incision enzyme driven multi-legged DNA molecular machine. The method comprises the following steps: aiming at target microRNA, designing and synthesizing detection probes 1, 2, 3 (probe 1, 2, 3) as recognition elements, designing and synthesizing a sulfhydrylation modified probe 4 (probe 4) as a report element, adding the sulfhydrylated probe 4 on the surface of a golden electrode and subjected to incubating overnight in a refrigerator at 4 DEG Cso that a report probe is anchored on the surface of the gold electrode and further a molecular orbit is prepared, identifying through a target sequence and a detection probe, and carrying out cascadeprogrammed reaction to form a trident DNA nano structure, designing a short nucleic acid restriction endonuclease site at each end of the structure as a leg of a molecular machine, and further forming the endonuclease driven multi-legged molecular machine. According to the present invention, the accurate temperature control PCR instrument is not required, the amplification can be achieved under the isothermal condition, and the advantages of simpleness, convenience and hypersensitivity are provided.

The invention relates to a long non-coding RNA for diagnosis, treatment and monitoring on the bladder cancer and application of the RNA. Through a lncRNA expression chip, an expression profile of lncRNA in a sample of a patient with the bladder cancer is detected, more than ten kinds of the lncRNA closely related to generation and development of the bladder cancer are discovered through analysis,the lncRNA is subjected to knock-down inhibition in a bladder cell line T24 in sequence, the drug sensitivity of T24 cells to DDP after all lncRNA sequences are inhibited is analyzed, a lncKMU15 whichis not reported at home and abroad is discovered, after knock-down, the IC500 of the DDP is obviously reduced, and the sensibility is obviously improved; the expression amount discovered through experimental verification is in obvious positive correlation with staging and grading of the tumor and in negative correlation with the total survival rate and non-tumor survival rate of the patient withthe bladder cancer, and the novel lncRNA-KMU15 provides brand-new possibility for exploration of a novel noninvasive molecular diagnosis technology to achieve sensitive early screening on the bladdercancer and an after-operation monitoring system.

The invention discloses a multiple touchdown PCR (polymerase chain reaction) detection kit capable of quickly detecting haemophilus influenzae in lower respiratory tract specimens, belongs to the technical field of molecular diagnosis of infectious diseases of lower respiratory tracts and aims to solve quick diagnosis of haemophilus influenzae in lower respiratory tract specimens. The kit comprises 10*PCR buffer solution, MgCl2, dNTP, TaqDNA polymerase, BSA (bovine serum albumin), haemophilus influenzae positive control DNA and three pairs of haemophilus influenzae primers. The invention discloses the multiple touchdown PCR detection kit and a detection method capable of detecting haemophilus influenzae and adopts agarose gel electrophoresis to detect the PCR products. The kit and the method have the advantages of quickness, simpleness and convenience, specificity and sensitivity and can be used for quick diagnosis and epidemiological survey of haemophilus influenzae infection.