77results about How to "Achieving absolute quantification" patented technology

The invention relates to application of differential expression of a gene in oral cancer diagnosis, and particularly relates to application of differential expression of LRRC26 gene in oral squamous cell carcinoma diagnosis. Transcriptome deep sequencing analysis is performed by virtue of a high-throughput sequencing platform to preliminarily screen the gene LRRC26 with remarkable differential depression in oral squamous cell carcinoma tissues, para-carcinoma tissues and normal tissues; and the RT-PCR experiment further proves that the LRRC26 gene has low expression with oral squamous cell carcinoma tissues, can be used for preparing oral squamous cell carcinoma auxiliary diagnosis or prognosis preparations, and has an important clinical practical application value.

The invention provides genes highly expressed in the tongue squamous carcinoma para-carcinoma tissue and applications of the genes, and in particular relates to the LCE3E gene and the GSDMA gene and applications of expression products of the LCE3E gene and the GSDMA gene in diagnosing tongue squamous carcinoma and preparing biological medicines for treating tongue squamous carcinoma. According to the invention, based on the high-throughput sequencing and in combination with the molecular biology experiment, the screened molecular diagnosis and treatment markers, namely, the LCE3E gene and the GSDMA gene for tongue squamous carcinoma are verified, therefore, an experimental foundation is provided for the clinical applications of the LCE3E gene and the GSDMA gene in tongue squamous carcinoma, and thus a novel target gene is provided for the early diagnosis and treatment of tongue squamous carcinoma.

The invention provides a self-driven microfluidic chip and a method for using the same. The self-driven microfluidic chip includes a substrate and a packaging sheet. A sample inlet, a sample outlet, aplurality of reaction chambers and one or more main channels are formed between the substrate and the packaging sheet. The sample inlet and the sample outlet can be communicated with the outside, themain channels are communicated with the sample inlet and the sample outlet, and liquid inlet channels and a liquid outlet channel are disposed between each of the reaction chambers and one main channel. The inner surfaces of the main channel, the reaction chambers, the liquid inlet channels and the liquid outlet channel located on the substrate and / or the packaging sheet are hydrophilic surfaces,the cross sectional area of the liquid inlet channel is smaller than that of the liquid outlet channel, and in the liquid flow direction in the main channel, the liquid inlet channels are located onthe front side of the liquid outlet channel. After a to-be-tested sample solution enters the reaction chamber, an oil phase reagent can be led into the main channel to seal the liquid inlet channels and the liquid outlet channel. The self-driven microfluidic chip can achieve self-driven flowing sample loading of the to-be-tested sample solution without an external driving device.

The invention relates to acute myocardial infarction related gene SERPINB13 and an application thereof. Through research, the inventor finds that the gene SERPINB13 has high expression in the peripheral blood of a patient suffering from the acute myocardial infarction, and meanwhile, the inventor designs interference RNAs of SERPINB13, wherein the interference RNAs can effectively inhibit the expression of the gene SERPINB13, and can be used for preparing the medicines for treating the acute myocardial infarction. With the adoption of the acute myocardial infarction related gene SERPINB13 and the application thereof, a novel solution is provided, and great application values are achieved.

The invention provides a kit and method for detecting the mutation of an EGFR gene and particularly discloses a method, a primer and a probe for detecting the mutation of the EGFR gene, a kit including prime probe mixing liquid, a primer and a probe for detecting the mutation of a C797S locus, a primer and a probe for detecting the mutation of a T790M locus, a primer and a probe for detecting themutation of a 19del locus and a primer and a probe for detecting the mutation of a L858 locus. According to the method provided by the invention, 0.1% mutation rate can be detected based on a digitalPCR platform; and the method has the advantages that the optimization process is simple, much detected mutation types can be detected, the absolute quantification is realized, the sensitivity is high,samples are easily acquired, and the like.

The invention provides a high-sensitivity EBV DNA quantitative detection kit based on ddPCR and a using method thereof. The kit comprises DNA extraction buffer solution, PCR primers and probes, a PCR reaction solution and EBV positive and negative control. When the kit provided by the invention is used for detecting ddPCR, the influences of PCR equipment, standard substances and the like can be effectively eliminated, and further high-sensitivity absolute quantification is further realized. The EBV DNA quantitative detection kit provided by the invention is capable of performing high-sensitivity accurate quantification on EBV in serum and plasma through a ddPCR technology, and providing a reference basis for diagnosis and curative effect monitoring of EBV related diseases. According to the kit, a function relationship between the international unit and common unit of EBV DNA international standard substances is determined to be 1IU / ml=2.5Copy / ml, the international traceability of EBV DNA is realized, and the international comparability between the detection results and quantification unit is realized.

The invention discloses a pair of primers and a probe for detecting hog cholera virus based on a digital PCR technology, a kit and a method thereof. The primers and the probe include an upstream primer FP, a downstream primer BP and a probe. The detection kit includes a primer set, 2*RT-ddPCR Supermix, RNase-free distilled water, a viral total RNA extraction reagent, a negative control and a positive control. The detection method comprises the following steps: extracting a sample RNA to be tested, quantitatively detecting the sample RNA to be detected by the microdrop digital PCR technique, judging whether the sample to be tested contains hog cholera virus by the copy number obtained by the amplification analysis, and determining the content thereof. The invention has the advantages of accuracy, sensitivity and wide applicability, has no dependency on a standard curve, can realize absolute quantification, can achieve early detection, early treatment and early prevention of diagnosis ofswine fever, and can effectively control the outbreak of the epidemic.

The invention provides a method and kit for quantitative detection of BCR-ABL fusion genes. The specific technical scheme comprises that two BCR-ABL fusion genetypes are detected only by two differentupstream primers, a common downstream primer, a fluorescent labeled probe and a set of ABL internal reference gene detection system. With use of the specific primers and probes provided by the invention, the two fusion types of BCR-ABL can be detected only by one tube of a material, and the kit has the advantages of high specificity, good sensitivity and high accuracy.

The invention belongs to the technical field of molecular biology and relates to a detection product for genomic methylation, in particular to a primer probe combination for methylation detection of an RASSF1A gene and a P16 gene and application of the primer probe combination. The primer probe combination for the methylation detection of the RASSF1A gene and the P16 gene comprises detection primer probes; and the detection primer probes comprise an RASSF1A gene methylation detection specific primer pair, an RASSF1A gene specific probe, a P16 gene methylation detection specific primer pair anda P16 gene specific probe. According to the primer probe combination, methylation sites of the RASSF1A gene and the P16 gene are accurately detected, detection precision of ctDNA is remarkably improved, and absolute quantification is realized; and in addition, the primer probe combination has high specificity and accuracy, and is suitable for early screening of liver cancer.

The invention belongs to the technical field of biomedical treatment, and specifically relates to a method and a test kit for detecting novel coronavirus. The test kit comprises a primer pair and a probe, wherein the sequences of the primer pair and the probe are as shown in SEQ ID NO. 1-6. The digital PCR detection test kit provided by the invention can reach the minimum detection sensitivity of50 copies / ml, realizes accurate quantification of trace pathogen nucleic acid, can detect pathogen nucleic acid from a large number of background signals, eliminates the dependence on a standard curve, does not need formula conversion and realizes absolute quantification of pathogen copies.

The invention relates to novel application of a transcription factor FOXD1, in particular to application of the transcription factor FOXD1 in regulating pituitary tumor disease. On the basis of a gene chip method, 10 pituitary tumor case samples and 5 normal comparison samples are sequenced, and a backup gene is screened. In addition, QPCR verification and siRNA interference experiment are performed, results show that the FOXD1 is highly related to pituitary tumor, and proliferation of pituitary tumor cells is slowed down by interfering FOXD 1 gene expression. The results provide a novel idea for clinical diagnosis and novel treatment of the pituitary tumor.

The invention provides an isotope derivatization reagent for labeling amino / phenolic hydroxyl and a synthesis method thereof and belongs to the field of chemical synthesis. A homologous compound of p-aminophenylacetic acid containing amino, a benzene ring and carboxyl in a structure or a homologous compound of 4-(2-aminoethyl)benzoic acid is used as a raw material; and the amino is dimethylated and then the carboxyl in the product is subjected to esterification by adopting N-hydroxysuccinimide to obtain the isotope derivatization reagent. The isotope derivatization reagent has structure characteristics that one part of the isotope derivatization reagent is succinimidyl ester and the other part of the isotope derivatization reagent is dimethyl substituted tertiary amino; the succinimidyl ester is connected to dimethylamino through a carbon chain and a benzene ring; and the number of carbon atoms on the benzene ring and the carbon chain which is connected to the amino is 0 to 6. According to the isotope derivatization reagent provided by the invention, different D substituted or <13>C substituted formaldehyde, or H or D substituted sodium cyanoborohydride are selected and a reactionroute is adjusted to obtain a series of isotope labeled reagents which can efficiently react with amino or phenolic hydroxyl and have different isotope substitutions. A starting raw material is changed, and the isotope labeled reagents with similar functions can be prepared through the method.

The invention provides a kit for miRNA detection and application thereof. The kit comprises a miRNA detection primer, DNA polymerase, incising endonuclease and a digital microfluidic chip. The kit isdeveloped based on the digital microfluidic chip and an isothermal amplification technology, and has relatively high detection sensitivity, specificity and accuracy. At the same time, the kit has relatively strong universality, and the detection of all miRNA can be realized only by designing a specific base sequence of a primer according to specific sequences of different miRNA. The kit provided by the invention directly can be used for directly detecting free and trace miRNA in body fluid without complicated pretreatment, RNA extraction, reverse transcription and the like on a human body fluid sample, is not interfered by other similar miRNA sequences, can realize absolute quantification of miRNA, has the advantages of simple operation, low requirement on detection equipment, low cost andthe like, and is suitable for wide popularization and application.

The invention discloses a centrifugal micro-droplet particle chip. The chip comprises an upper piece and a lower piece bonded with the upper piece, a sample storage ring arranged in the middle of theouter surface of the upper piece or in the middle of the outer surface of the lower piece, M closed sample flow channels which are formed in a position between the upper piece and the lower piece andare outward distributed in a radial manner from the middle of the chip, and closed arc micro-droplet particle storage slot groups (5) which are arranged between the upper piece and the lower piece onthe left sides of the flow channels, wherein M is greater than 2; the closed arc micro-droplet particle storage slot groups comprise a plurality of arc micro-droplet particle storage slots which sharethe same center of a circle; M oil discharge flow channels which are outward distributed in a radial manner from the middle of the chip are formed in a position between the lower surface of the upperpiece and the upper surface of the lower piece; each oil discharge flow channel extends to the edge of the chip and is communicated with an oil discharge outlet at the edge of the chip. The chip hasthe characteristics of high sensitivity and high accuracy.

The invention relates to a PDMS material-based microchamber array type digital PCR chip. The chip is made of a PDMS material and comprises a sample inlet hole, a microchamber array and a sample outlet hole; the sample inlet hole comprises a sample inlet and a water sample inlet; the sample outlet hole comprises a sample outlet and a water sample outlet; the water sample inlet and the water sample outlet are communicated with the microchamber array at the periphery of the chip; and the sample inlet and the sample outlet are communicated with each reaction chamber in the microchamber array through a microchannel. According to the PDMS material-based microchamber array type digital PCR chip, water channels at the periphery of the chip can prevent PCR reaction liquid in the PCR process from evaporating; a designed automatic counting software can automatically counts a chip result; and the PDMS material-based microchamber array type digital PCR chip can reach very high detection sensitivity and realizes single molecule detection of DNA.

The invention discloses a test kit and a detection method for quantitative detection of a high-throughput sequencing library. The test kit comprises a detection primer and a detection probe which aredesigned for the joint sequence of a to-be-detected high-throughput sequencing library, wherein upstream and downstream primers of the detection primer have sequences shown as Seq ID No.1 and Seq ID No.2; the detection probe has a sequence shown as Seq ID No.3; the 5'end of the detection probe is modified by a fluorophore; and the 3'end of the detection probe is modified by a fluorescence quenching group. According to the test kit, an NGS library is quantitatively detected through a TaqMan probe method, and absolute quantification of the NGS library can be achieved; and compared with a conventional dye method for QPCR quantitative detection, the test kit can obtain the actual concentration of the NGS library more accurately; the size of a DNA fragment based on the NGS library does not needto be corrected, which is of great significance to high-throughput sequencing; and the sequencing quality can be improved, so an optimal sequencing data result is obtained.

The invention relates to new application of a KCNK2 gene, in particular to application of the KCNK2 gene and an expression product thereof to treatment of papillary thyroid carcinoma as tumor markers .In order to diagnose thyroid carcinoma more accurately and avoid missed diagnosis and misdiagnosis, tumor molecular markers are screened based on sequencing and data analysis, the result shows that the KCNK2 gene can be used for preparing a papillary thyroid carcinoma auxiliary diagnosis and treatment preparation, and quite good clinical application value is achieved.

The invention discloses a primer and probe, a kit and a method for detecting streptococcus suis based on the digital PCR technology. The primer and probe include a forward primer FP, a backward primerBP and a Probe. The detection kit includes 2 x ddPCR Supermix, ultrapure water without RNA enzymes, a bacterial total DNA extraction reagent, negative control and positive control. The detection method comprises the steps that DNA of a sample to be tested is extracted, the DNA of the sample to be tested is quantitatively detected by the microdrop digital PCR technology, a copy number obtained byamplification analysis is used for determining whether the sample to be tested contains the streptococcus suis or not, and the content of the streptococcus suis is determined. The primer and probe, the kit and the method for detecting the streptococcus suis based on the digital PCR technology have the advantages of accuracy, sensitivity, wide applicability, no dependence on a standard curve, realization of absolute quantification and the like, can realize early detection, early treatment and early prevention of the diagnosis of the streptococcus suis disease, and can effectively control the disease outbreak.

A DNA methylation degree quantifying method and application are provided. The method includes (1) adding sulfite modified DNA into a PCR system, and performing digital PCR; and (2) acquiring the DNA methylation degree according to the DNA methylation copy number and non-methylation copy number. The PCR system in the step (1) includes a Taqman probe, with the Taqman probe including a methylation Taqman probe or a non-methylation Taqman probe. The target gene modified with sulfite is detected with the methylation probe and the non-methylation probe separately, and the methylation copy number andthe non-methylation copy number of the gene to be detected are measured by the digital PCR accurately, thus achieving absolute quantification of the methylation degree of the target gene. The methodis suitable for detection of trace ctDNA in blood samples, is high in sensitivity, high in specificity, precise and rapid, and facilitates promotion in the technical field of liquid biopsy.

The invention relates to application of an SPAG7 (Sperm Associated Antigen) gene in preparing a senile dementia diagnosis kit, in particular to application of the SPAG7 gene and an expression productthereof in diagnosing and treating senile dementia diseases, and aims to solve the problem that senile dementia molecular markers are scarce at present. According to the application, peripheral bloodsamples of patients suffering from senile dementia and reference peripheral blood samples of healthy people are subjected to high-flux sequencing, the candidate gene SPAG7 is selected, an RT-PCR (Reverse Transcription-Polymerase Chain Reaction) method shows that the SPAG7 gene has very good correlation with senile dementia, and a base is made for clinical gene diagnosis on senile dementia.

The invention belongs to the field of detection of virus nucleic acid, and particularly relates to a digital PCR reagent kit and detection method for quantitative detection of EB virus nucleic acid. The reagent kit contains digital PCR reaction liquid and a quality control product, wherein the digital PCR reaction liquid contains a primer and probe for detecting EB virus nucleic acid, and comprises a forward primer as shown in SEQID NO:1, a reverse primer as shown in SEQID NO:2 and a probe as shown in SEQID NO:3. Fluorophore is marked at two ends of the probe. The reagent kit and the detectionmethod can realize absolute quantitation of nucleic acid molecules, so as to avoid deviation caused by PCR amplification efficiency variance, and the reagent kit and the detection method have high accuracy, sensitivity and repeatability, and easily realize standardization.

The invention belongs to the technical field of food safety, and discloses an intelligent food quality fast detection system. The intelligent food quality fast detection system is provided with a foodsafety system control end; the food safety system control end is connected with an intelligent food classification instrument through a data wire; the intelligent food classification instrument is connected with an aquatic product detection module, a meat detection module, a fruit and vegetable detection module and an other food monitoring module; the aquatic product detection module, the meat detection module, the fruit and vegetable detection module and the other food monitoring module are connected with an enzyme linked immunosorbent assay detection module; the enzyme linked immunosorbentassay detection module is connected with a pesticide residue compound detection module, a microorganism module, a fluorescent quantitative PCR module and a food attribution module through data wires.According to the intelligent food quality fast detection system provided by the invention, the food is classified, so that the food quality detection is faster; through the enzyme linked immunosorbentassay detection module, an antibody and an enzyme compound are combined; whether food is transgenic food or not is detected through color development; the food health and safety are ensured, so thatconsumers are relieved.

The invention relates to a droplet digital PCR (Polymerase Chain Reaction) kit for detecting carp edema virus. The kit includes a primer group and a fluorescent probe. The primer group includes an upstream primer of the sequence shown in SEQ ID NO.8 and a downstream primer of the sequence shown in SEQ ID NO.9. The fluorescent probe includes the fluorescent probe sequence shown in SEQ ID NO.5. Whenusing the kit to perform droplet digital PCR, the lowest detection limit of CEV (carp edema virus) is 1.8 copies / muL which is better than the sensitivity (17.6 copies / muL) of the fluorescent quantitation PCR detection method of the CEV; and no cross reaction is found with the nucleic acids of important aquatic animal pathogens like andriasda vidianus ranaviurs (ADRV), bohle virus (BIV), channel catfish virus (CCV) and spring viremia of carp virus (SVCV). The droplet digital PCR kit provided by the invention is of good specificity.

The invention provides a high-throughput sequencing method for absolute quantification of RNA molecules. The method comprises the following steps: dephosphorylating a RNA sample; ligating the dephosphorylated RNA to DNA linkers 1 by T4 RNA ligase to capture 95% or more than 95% of RNA; then using demethylase to reduce RNA modification; removing excess linkers 1; reverse-transcribing to obtain cDNAwith the RNA as a template; degrading the RNA template; ligating the cDNA to DNA linkers 2 of a hairpin structure by the T4 DNA ligase so as to ligate 98% or more than 98% of the cDNA; removing excess linkers 2; and finally performing PCR primer amplification and sequencing. According to the invention, the defects of inaccurate quantification caused by the ligation efficiency preference in the existing RNA sequencing technology and the inability to capture RNA molecules containing modified sites are overcome by specially designing the DNA linkers, thereby realizing the absolute quantificationof all RNA molecules in RNA samples from different sources. The method provided by the invention is simple and can be used in the fields of clinical molecular diagnosis, molecular biological research, bacterial resistance, cancer occurrence and prevention and the like.

The technical solution of the present invention discloses an absolute quantification analysis method of ceramide. After a to-be-tested sample, a working reagent and a solvent are uniformly mixed, a mixture is stratified to form an upper-layer to-be-tested sample and a lower-layer to-be-tested sample, the upper-layer to-be-tested sample and a calibration reagent are detected by using a high performance liquid chromatography-tandem mass spectrum system, to obtain a mass spectrum detection result of the calibration reagent, calibration is performed according to mass spectrum detection results ofat least two calibration reagents, and absolute quantification analysis is performed on the ceramide in the to-be-tested sample according to a calibration result and a mass spectrum detection result of the upper-layer to-be-tested sample. According to the analysis method provided in the technical solution of the present invention, molecular number absolute quantification on ceramide can be effectively performed by using only a small quantity of serum.

The invention provides a kit for detecting activity degree of telomerase in a whole blood or serum sample, belonging to the field of molecular biology detection. The kit comprises the following primers: (1) primer sequences of telomerase hTERT genes: hTERT-F: 5'-GGCCGATTGTGAACATGGA-3'; hTERT-R: 5'-CCTCTTTTCTCTGCGGAACGT-3'; and (2) probe sequences of telomerase hTERT genes: hTERT-P: 5'-TACGTCGTGGGAGCCA-3', the 5' end is connected with fluorescence reporter genes, and the 3' end is connected with fluorescence quenching genes. The kit for detecting activity degree of telomerase in the whole blood or serum sample has the advantages of high detection sensitivity, short analysis time and high stability.

The invention provides a method for detecting bile acid in excrement by UPLC-MS / MS. The method comprises the following steps: establishing a standard curve; treating a to-be-detected excrement sample,and dissolving the frozen to-be-detected excrement sample by adopting mass spectrum water to obtain to-be-detected slurry; carrying out extraction and centrifugal separation on the to-be-detected slurry by adopting an extracting agent to obtain supernatant; drying the supernatant, and performing redissolving by adopting an initial mobile phase to obtain a redissolving system; centrifuging the redissolving system to obtain a to-be-detected solution, wherein the extracting agent comprises a mixed solution of acetonitrile and methanol, the volume ratio of acetonitrile to methanol in the mixed solution is 7:3-9:1, and the initial mobile phase comprises a mixed solution of acetonitrile and a 0.1% formic acid aqueous solution in a volume ratio of 7:3-9:1; detecting the to-be-detected solution by adopting UPLC-MS / MS to obtain ion spectrum peaks of bile acids; and converting the content of each bile acid according to the standard curve and the ion spectrum peak. The method can be used for quantifying 33 bile acids.

The invention relates to the field of biomedicine, in particular to LYRM2 (LYR motif-containing 2) gene and new use of an expression product thereof. In order to solve the problem that osteoporosis molecular markers are scarce at present, the inventor screens pathogenic target genes based on the analysis of high-throughput sequencing in connection with bioinformatic methods, selects candidate gene LYRM2, verifies that LYRM2 gene is well related to osteoporosis by molecular biology experiments, and also designs an efficient interference RNA for LYRM2. The osteoporosis assisted diagnosis and treatment target provided by the invention is well worthy of clinical application.

Belonging to the technical field of fluorescence in-situ hybridization detection, the invention relates to an absolute quantification method for fluorescence in-situ hybrid cell counting. The absolute quantification method mentioned in the invention adds a weight quantification step in a fluorescence in-situ hybridization process, makes improvements to sampling, sample pretreatment, experimental treatment, late-stage result analysis and other stages of an original experimental scheme, so that the complete set of method is established. The method is characterized in that: when quantifying a biological membrane and other solid samples, a quantitative high speed centrifuged sample can be adopted, high temperature drying is performed on the same sample at the same time, then the sample is subjected to fluorescence in situ hybridization and observation under a microscope as well as calculation, thus finally obtaining the microorganism concentration of dry sludge with certain weight. The method realizes comparison of different forms of samples in a same reactor in terms of microbial community numbers, and at the same time, puts forward an absolute quantification means able to establish a standard method.

The invention relates to a digital PCR method for detecting the brucella nucleic acid DNA, and relates to the technical field of molecular biology. A digital PCR technology in the invention can detectand identify pure brucella nucleic acid DNA, and also can detect and identify trace brucella nucleic acid DNA in tissue fluid, such as blood; the brucella nucleic acid DNA of a sample to be detectedcan be qualitatively identified; the copy number of the brucella nucleic acid DNA of the sample to be detected also can be quantitatively detected; the minimum 3.67 fg / [mu]L of the nucleic acid DNA can be detected; namely, the minimum detectable range can be one brucella nucleic acid DNA copy; in addition, digital PCR result analysis can be standardized; and data comparison of different experimenters can be carried out.