31 results about "Quantitative Real-Time Polymerase Chain Reaction" patented technology

The present invention provides a novel robust, sensitive, reproducible, and accurate method of detecting and quantifying host cell genomic DNA contamination utilizing quantitative real time Polymerase Chain Reaction (qPCR), wherein the qPCR primers are complementary to the highly repetitive host cell genomic DNA sequences, e.g., Alu-equivalent sequences. The present invention is particularly useful for determining the levels of residual genomic DNA in biological products to be administered as therapeutics, e.g., therapeutic proteins.

The invention relates to the technical field of the biological experimental facilities and discloses a coordinate sample injection box of a 384-hole PCR plate for quantitative real-time polymerase chain reaction. The coordinate sample injection box of the 384-hole PCR plate for the quantitative real-time polymerase chain reaction comprises a sample injection box and a base, wherein the sample injection box is a shape like a cuboid with only four side surfaces; the sample injection box is matched with the 384-hole PCR plate in size; the 384-hole PCR plate can be placed in the sample injection box; a movable transverse mark post and a movable longitudinal mark post are arranged on the sample injection box; 1 to 24 figure marks are formed on the transverse mark post, while A to P letter marks are formed on the longitudinal mark post, and each figure and each letter are capable of corresponding to one micropore in the 384-hole PCR plate; the base is used for placement of the 384-hole PCR plate and the sample injection box and can be separated from the sample injection box. The coordinate sample injection box of the 384-hole PCR plate for the quantitative real-time polymerase chain reaction is capable of realizing accurate positioning of every micropore in the 384-hole PCR plate and immediately tracking the position of the sample injection micropores, and is capable of stopping in one position any time without error.

The present invention relates to a method for detection, differentiation and quantification of T cell populations, comprising the following steps a) contacting a first aliquot of a body fluid of an individual with at least one antigen, wherein the body fluid contains antigen presenting cells (APC) and T cells, b) incubating the first aliquot with at least one antigen for a certain period of time, c) detection and differentiation of the T cell population by detecting in the first aliquot and in a second aliquot of the body fluid of the individual, which has not been incubated with the at least one antigen, at least a first marker of the APC induced by T cells in a specific T cell population using reverse transcription quantitative real time-time polymerase chain reaction (RT-qPCR), and d) detection and quantification of the T cell population by determining the ratio of the detected marker of the APC of the first aliquot to the second aliquot as well as a kit for performing the method.

The invention relates to long non-coding RNA and application thereof. According to a sequence of the long non-coding RNA, specific qRT-PCR (Quantitative Real-time Polymerase Chain Reaction) primers and probes are designed and synthesized and reagents used for esophageal carcinoma auxiliary diagnosis or curative effect prediction are prepared. The qRT-PCR reagents are utilized for detecting the expression level of the long non-coding RNA in specimens of esophageal carcinoma clinical cases and a substantial decrease of the long non-coding RNS expression is found in esophageal carcinoma. The long non-coding RNA can be applied to preparing reagents used for esophageal carcinoma auxiliary diagnosis, curative effect prediction or prognosis judgment.

The invention relates to the field of the molecular biology and provides an improved method for extracting total RNA in each growth period of pichia pastoris, aiming to solve the problem of difficult extraction of total RNA of pichia pastoris cells occuring in molecular research. The method is improved on the basis of Sangon UNIQ-10 column type Trizol total RNA extraction kit, and suitable for extraction of total RNA in each growth period of pichia pastoris in contrast with methods for extracting total RNA by using hot acid phenol; even for the stable-phase cells having thick cell walls, the method is still capable of successfully extracting total RNA and the extraction result is stable; moreover, the total RNA extracted by the method is good in quality, and has clear and tidy 28SrRNA and 18SrRNA strips in 2-time relationship; besides, the method is capable of meeting the purity requirement between 1.8 and 2.1 of OD260 / OD280; and degradation hardly occurs; and therefore, the method meets the requirement of QRT-PCR (quantitative real-time polymerase chain reaction) measurement.

The invention discloses fluorescence quantitation internal reference genes of leaf tissues of different parts of two pear trees different in three form and primer and application thereof. According tothe fluorescence quantitation internal reference genes of leaf tissues of different parts of the two pear trees different in three form and primer and application thereof, qRT-PCR (quantitative real-time polymerase chain reaction) is applied to analyzing the expression level of 13 candidate internal reference genes in pear leaf tissue samples of different parts of the two pear trees different intree form, geNorm software is applied to estimating the expression stable value of the candidate internal reference genes to determine that ARM and Actin2 are the optimal internal reference gene combination; different internal reference genes are applied to performing relative expression standardization analysis on APX to prove that the internal reference combination of ARM and Actin2 is high inreliability when serving as normalized factors.

The invention relates to esophageal carcinoma-related long non-coding RNA and application thereof. Based on the sequence of the long non-coding RNA, specific qRT-PCR (Quantitative Real-time Polymerase Chain Reaction) primers and probes are designed and synthesized and reagents used for esophageal carcinoma auxiliary diagnosis or curative effect prediction are prepared. The qRT-PCR reagents are utilized for detecting the expression level of the long non-coding RNA in specimens of esophageal carcinoma clinical cases and a substantial decrease of the long non-coding RNS expression is found in esophageal carcinoma. The esophageal carcinoma-related long non-coding RNA can be applied to preparing reagents used for esophageal carcinoma auxiliary diagnosis, curative effect prediction or prognosis judgment.

The present disclosure relates to a highly specific and sensitive method of detecting host cell impurities in a biological sample by using quantitative real time polymerase chain reaction (q PCR). The present disclosure also provides novel designed primer and probe to amplify only the specific Alu family of dispersed repetitive sequences from Chinese hamster ovary cells used for expression of therapeutic proteins.

The invention discloses application of application of OC-STAMP (osteoclast stimulatory transmembrane protein) as a prognostic risk marker in evaluating multiple myeloma patients. According to the application, expression levels of OC-STAMP in bone marrow cells of newly diagnosed and released MM (multiple myeloma) patients and healthy donors are analyzed by using an RT-qPCR (real-time-quantitative real time polymerase chain reaction) method, and results show that the expression level of the gene in an MM patient is remarkably higher than that of a normal reference level and is possibly related to clinical courses. Subsequently, the clinical significance of the gene is investigated and results show that high expression of the gene is an independent risk factor of low progression-free survival(PFS) of the MM patient. OC-STAMP has important value in evaluating the prognostic risk of the multiple myeloma patients.

The present invention relates to methods and uses for the detection or quantification of newly-synthesized double-stranded target nucleic acid molecules in a sample during quantitative real-time polymerase chain reaction (qPCR) amplification. According to the invention, an intercalating dye recognizing double-stranded DNA molecules with higher affinity than single-stranded DNA molecules and a fluorophore-labeled oligonucleotide-probe being sequence specific for a target nucleic acid molecule are simultaneously employed, thus enabling quantification a specific target and total amount of a mixed nucleic acid population, and enabling assessing the cause of suboptimal PCR performance.

The invention provides an application of a PPAR alpha-UGT (peroxisome proliferator activated receptor alpha-uridine diphosphate glucuronosyl transferase) pathway inhibitor in treating inflammatory bowel diseases, belongs to the field of new medicine research and development, and in particular relates to discovery of the treatment effect of the PPAR alphas-UGT pathway inhibitor, which is used for relieving hepatic and gall disease complications of inflammatory bowel diseases by controlling the steady equilibrium of bile acid, and reducing the risk that inflammatory bowel diseases are transformed to colon cancer. The application proves that the PPAR alpha-UGT pathway can be remarkably activated in a DSS colonitis mouse due to the detection to mice in intestine PPAR alpha and downstream typical target genes of a normal group and DSS (dextran sodium sulfate) induced colonitis mice by a qRT-PCR (quantitative real-time polymerase chain reaction) technology; and the application discovers and proves that an FXR-FGF15 (farnesoid X receptor-fibroblast growth factor15) negative feedback pathway of bile acid is remarkably inhibited in the colonitis mouse and bile acid synthesis is remarkably increased due to the qRT-PCR technology and an immunohistochemical technology. Therefore, activation of the PPAR alpha-UGT pathway is an initial factor of homeostasis imbalance of bile acid in inflammatory bowel diseases, and the PPAR alpha-UGT pathway inhibitor is expected to become a new target for researching and developing medicines for treating vicious transformation of inflammatory bowel diseases and complications thereof.

The invention discloses a new application of ligusticum chuanxiong hort or an extract thereof. The ligusticum chuanxiong hort or the extract thereof has the following four new purposes: (1) application in preparing anti-aging products; (2) application in preparing gene expression products for regulating caloric restriction pathways; (3) application in preparing gene expression products for regulating insulin signaling pathways; and (4) application in preparing caenorhabditis elegans life extenders. In the invention, the caenorhabditis elegans is used as a model organism, the influence of CXE (Ligusticum Chuangxiong Extract) on caenorhabditis elegan life is observed by taking a whole animal experiment as a detection means, the CXEs of a plurality of concentrations are all found to have a role or trend of life prolongation to the caenorhabditis elegan, wherein the concentration of 25mg / L has an optimal life prolongation effect, and the average life and the longest life are both prolonged by about 15 percent. Molecular mechanism research for life prolongation by utilizing the known relevant caenorhabditis elegan aging gene and adopting a qRT-PCR (Quantitative Real Time-Polymerase Chain Reaction) method shows that the CXE prolongs the life through intervening an Insulin / IGF-1 pathway and a feeding control (caloric restriction) pathway, thereby providing new content and direction for the molecular mechanism research of the drug and also providing a new ideal and means for the research of delaying aging by using the traditional Chinese medicine.

The invention provides application of molecule miR-150 in screening preparation of a colorectal cancer detecting medicament. The molecule miR-150 is screened out by using a muParaflo microfluidic chip by the inventor of the invention; a paraffin section preparation of a clinical colorectal cancer patient is detected by an established digoxigenin labeled probe of miR-150 and an established detection method of qRT-PCR (Quantitative Real-Time-Polymerase Chain Reaction), respectively; results show that miR-150 low expression level in the clinical colorectal cancer patient indicates short lifetime of the patient and poor sensitivity of the patient to postoperative chemotherapy, and miR-150 high expression level indicates long lifetime of the patient and good sensitivity of the patient to postoperative chemotherapy. Compounds having high sensitivity to miR-150 can be screened by the method, and the compounds can be used for developing a kit for detecting the colorectal cancer. The kit is composed of compounds sensitive to the molecule miR-150 as active ingredients and a medicinal carrier. Simple detection of the miR-150 expression level is advantageous for monitoring of the colorectal cancer patient after healing and determination of chemotherapy sensitivity of the patient, and thus provides an effective method for clinical prevention and treatment of the colorectal cancer; high clinical application value and social benefit are created.

The invention discloses a detection primer and a probe for rapid identification of sturgeon based on quantitative real time polymerase chain reaction (PCR) as well as a detection reagent kit and a detection method which adopt the primer and the probe. A detection primer group comprises a forward primer FP, a backward primer BP and the probe; the detection reagent kit comprises a primer solution, aPCR mix reaction solution, deionized water, a negative control and a positive control; and the detection method comprises the following steps: the DNA of a sample to be examined is extracted; the quantitative PCR is utilized for rapidly identifying the DNA of the sample to be examined; and whether the sample to be examined is the sturgeon or not is judged by a real-time amplification curve. The invention has the advantages of high speed, sensitivity, specificity, easy operation, and the like, and is suitable for popularization and application.

The invention relates to a method for detecting the yield of royal jelly by using a bee LOC726515 gene. The method comprises bee sample collection, bee head RNA (Ribose Nucleic Acid) extraction, bee head cDNA (complementary Deoxyribonucleic Acid) synthesis, primer design, Qrt-PCR (quantitative Real-Time Polymerase Chain Reaction) reaction system and conditions, and royal jelly yield performance identification. The invention provides a fluorescent quantitative PCR method which is capable of identifying good or bad royal jelly yield performance of a swarm of bees more scientifically and accurately at the molecular biology level, aiming at the problem that the production of the royal jelly is enormously affected by conditions such as the population of the swarm of bees, nectar sources and weather, and as a result, the study period of the royal jelly production traits can be greatly shortened, the honeybee breeding speed can be increased, the cost of studying the royal jelly production traits and the cost of breeding can be reduced, and the working strength of the field study of the royal jelly yield performance also can be reduced.

The invention discloses an establishing method and application for a multi-gene liver cancer prognosis grading system. The system can be used to predict survival time and recurrence probability of patients with liver cancer after surgery, regardless of the ages and tumor sizes of the patients. Samples come from a total of 378 clinical liver cancer surgery patients in a hospital affiliated to Zhejiang University School of Medicine in 2012-2018, after strict quality control of the samples, through a multi-gene quantitative real time polymerase chain reaction (qRT-PCR), the correlation between each gene and the survival time is further analyzed, and finally high-risk genes related to prognosis of the clinical liver cancer surgery patients are screened to establish the prognosis grading system. The method has the following beneficial effects: the prognosis grading system capable of predicting the survival time and recurrence probability of the clinical liver cancer patients after the surgery is established, regardless of the ages and tumor sizes of the patients, and the system can be used for postoperative survival time assessment of the liver cancer surgery patients, and assist in formulation of a postoperative treatment plan for the patients.

The invention relates to a method of detecting yield of royal jelly by LOC409360 gene of a bee. The method comprises the following steps: bee sample collection; extraction of RNA (Ribonucleic Acid) at the head of the bee; synthesis of cDNA (Complementary Deoxyribonucleic Acid) at the head of the bee; primer designing; qRT-PCR (Quantitative Real-Time Polymerase Chain Reaction) reaction system and condition; and yield capability identification of royal jelly. In the level of molecular biology, the invention provides a fluorescent quantitative PCR method which can more scientifically and accurately identify the yield capability of a swarm in order to solve the problem that production of royal jelly is hugely affected by conditions such as trend of the swarm, nectar sources, environment and climate, thereby greatly shortening the inspection period of production trait of royal jelly, accelerating the breeding speed of bees, reducing the inspection cost and breeding cost of production trait of royal jelly and further alleviating the working strength of inspection of yield of royal jelly in the field.

The invention provides qRT-PCR (quantitative real-time polymerase chain reaction) for detecting the RBSDV (rice black-streaked dwarf virus) and an application thereof, belonging to the technical field of PCR detection. Firstly, a conserved segment is determined by comparing genomic sequences of the RBSDV and a primer pair and probe used for detecting the RBSDV are designed by using the conserved region as a target sequence. The nucleotide sequences of the primer pair and probe are respectively shown in SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.3. The invention also provides a method and detection kit for carrying out real-time fluorescence quantitative detection on the RBSDV. The detection method has the advantages of accuracy in detection, high sensitivity, strong specificity, simplicity and quickness, can achieve quantitative detection of early infection of crops with the RBSDV, is low in cost and has good economical value and application prospect.

Disclosed herein are methods, compositions and kits for the quantification of a nucleic acid target present on a solid support. This entails quantitative real-time polymerase chain reaction wherein minor groove binders are excluded.

The invention relates to the technical field of medical biological detection and provides an HOST2 detection kit for assisting in early diagnosis of an epithelial ovarian cancer (EOC). The HOST2 detection kit consists of a reverse transcription system, a primer system and an amplification system, wherein HOST2 specific qRT-PCR (quantitative real-time polymerase chain reaction) upstream primer is shown as SEQ ID No: 2; and a downstream primer is shown as SEQ ID NO: 3. The kit adopts a qRT-PCR method, detects an HOST2 content of plasma or serum for assistance of the early diagnosis of the EOC, and has the characteristics of small trauma, high operability and the like. The invention further provides a detection method of the HOST2 detection kit for assisting in the early diagnosis of the EOC.

The invention discloses an application of UCP1 (uncoupling protein 1) to activity detection of human recombinant FGF21 (fibroblast growth factor 21) protein. Adipocytes subjected to induced differentiation are taken as objects, qRT-PCR (quantitative real-time polymerase chain reaction) is adopted to detect the rising degree of UCP1 gene expression level of the adipocytes processed by the human recombinant FGF21 protein with different concentrations, and the activity of the human recombinant FGF21 protein is determined. The invention discovers that the UCP1 expression level and the concentration of the used human recombinant FGF21 protein have a good dose-effect relationship, so that the UCP1 expression level of the adipocytes is determined to be a new index for activity detection of the human recombinant FGF21 protein. A method adopts a simple operation process, has low environmental requirement and has good practical value.

Disclosed herein are methods, compositions and kits for the quantification of a nucleic acid target present on a solid support. This entails quantitative real-time polymerase chain reaction wherein minor groove binders are excluded.

The invention discloses a method for establishing a polygenic liver cancer prognosis grading system and its application, which can be used to predict the survival period and recurrence probability of liver cancer patients after surgery, regardless of the patient's age and tumor size. The samples came from a total of 378 clinical patients with liver cancer surgery in an affiliated hospital of Zhejiang University School of Medicine in 2012-2018. After strict quality control, the samples were subjected to multi-gene quantitative real-time polymerase chain reaction (qRT-PCR) , and further analyzed the correlation between each gene and the survival period, and finally screened out the high-risk genes related to the prognosis of patients with clinical liver cancer surgery, and constructed a prognosis grading system. Beneficial effects of the patented technology of the present invention: the present invention constructs a prognosis grading system that can predict postoperative survival and recurrence probability of patients with clinical liver cancer, regardless of patient age and tumor size. It can be used to evaluate the postoperative survival of patients with liver cancer surgery, and assist in formulating postoperative treatment plans for patients.

The invention relates to an antrodia cinnamomea housekeeping gene sequence. The housekeeping gene sequence refers to a c4163 or c10657 sequence. The c4163 sequence is shown as SEQ ID NO: 1. The c10657 sequence is shown as SEQ ID NO: 2. The invention further relates to a screening method of the antrodia cinnamomea housekeeping gene sequence, a qRT-PCR (quantitative real-time polymerase chain reaction) amplification primer of the antrodia cinnamomea housekeeping gene sequence and a qRT-PCR amplification method of the antrodia cinnamomea housekeeping gene sequence. The antrodia cinnamomea housekeeping gene sequence can be stably expressed in antrodia cinnamomea samples with different shapes and cultivated in different times, stripe singleness is achieved, and a correction sequence is provided for research on functional gene expression in antrodia cinnamomea.

The invention discloses a new application of ligusticum chuanxiong hort or an extract thereof. The ligusticum chuanxiong hort or the extract thereof has the following four new purposes: (1) application in preparing anti-aging products; (2) application in preparing gene expression products for regulating caloric restriction pathways; (3) application in preparing gene expression products for regulating insulin signaling pathways; and (4) application in preparing caenorhabditis elegans life extenders. In the invention, the caenorhabditis elegans is used as a model organism, the influence of CXE (Ligusticum Chuangxiong Extract) on caenorhabditis elegan life is observed by taking a whole animal experiment as a detection means, the CXEs of a plurality of concentrations are all found to have a role or trend of life prolongation to the caenorhabditis elegan, wherein the concentration of 25mg / L has an optimal life prolongation effect, and the average life and the longest life are both prolonged by about 15 percent. Molecular mechanism research for life prolongation by utilizing the known relevant caenorhabditis elegan aging gene and adopting a qRT-PCR (Quantitative Real Time-Polymerase Chain Reaction) method shows that the CXE prolongs the life through intervening an Insulin / IGF-1 pathway and a feeding control (caloric restriction) pathway, thereby providing new content and direction for the molecular mechanism research of the drug and also providing a new ideal and means for the research of delaying aging by using the traditional Chinese medicine.

The invention relates to long non-coding RNA and application thereof. According to the sequence of the long non-coding RNA, specific qRT-PCR (Quantitative Real-time Polymerase Chain Reaction) primers and probes are designed and synthesized and reagents used for esophageal carcinoma auxiliary diagnosis or curative effect prediction are prepared. The qRT-PCR reagents are utilized for detecting the expression level of the long non-coding RNA in specimens of esophageal carcinoma clinical cases and a substantial decrease of the long non-coding RNS expression is found in esophageal carcinoma. The long non-coding RNA can be applied to preparing reagents used for esophageal carcinoma auxiliary diagnosis, curative effect prediction or prognosis judgment.

The invention relates to the technical field of the biological experimental facilities and discloses a coordinate sample injection box of a 384-hole PCR plate for quantitative real-time polymerase chain reaction. The coordinate sample injection box of the 384-hole PCR plate for the quantitative real-time polymerase chain reaction comprises a sample injection box and a base, wherein the sample injection box is a shape like a cuboid with only four side surfaces; the sample injection box is matched with the 384-hole PCR plate in size; the 384-hole PCR plate can be placed in the sample injection box; a movable transverse mark post and a movable longitudinal mark post are arranged on the sample injection box; 1 to 24 figure marks are formed on the transverse mark post, while A to P letter marks are formed on the longitudinal mark post, and each figure and each letter are capable of corresponding to one micropore in the 384-hole PCR plate; the base is used for placement of the 384-hole PCR plate and the sample injection box and can be separated from the sample injection box. The coordinate sample injection box of the 384-hole PCR plate for the quantitative real-time polymerase chain reaction is capable of realizing accurate positioning of every micropore in the 384-hole PCR plate and immediately tracking the position of the sample injection micropores, and is capable of stopping in one position any time without error.

The invention provides qRT-PCR (quantitative real-time polymerase chain reaction) for detecting the RBSDV (rice black-streaked dwarf virus) and an application thereof, belonging to the technical field of PCR detection. Firstly, a conserved segment is determined by comparing genomic sequences of the RBSDV and a primer pair and probe used for detecting the RBSDV are designed by using the conserved region as a target sequence. The nucleotide sequences of the primer pair and probe are respectively shown in SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.3. The invention also provides a method and detection kit for carrying out real-time fluorescence quantitative detection on the RBSDV. The detection method has the advantages of accuracy in detection, high sensitivity, strong specificity, simplicity and quickness, can achieve quantitative detection of early infection of crops with the RBSDV, is low in cost and has good economical value and application prospect.