222 results about "Quantitative PCR instrument" patented technology

The invention discloses a PCR (Polymerase Chain Reaction) chip based on droplet array. The chip is obtained by taking a mono crystalline silicon sheet or a glass sheet as a base material and adopting standard photoetching as well as wet etching technique. The mono crystalline silicon sheet or the glass sheet is the base material of the chip. A SiO2 oxide layer and a silane layer generated by silanization on the surface of the SiO2 oxide layer are adhered to the base material successively. A hydrophilic pit array, which can be a droplet array and is formed by etching with photoresist technology, is disposed on the silane layer. A closed circular guardrail is arranged on the outer periphery of the hydrophilic pit array and the guardrail is connected with the silane layer in a sealed manner. The invention also discloses the application of the chip on real-time quantitative PCR detection and real-time quantitative isothermal amplification reaction detection. The detection system provided in the invention remarkably reduces the reaction volume and the detection sensitivity of the system fairly equals that of a commercialized quantitative PCR instrument. The detection system in the invention is easy for operation and has low cost, thus can be applied to commercialization.

The invention discloses a fluorescence quantitative PCR (Polymerase Chain Reaction) detection method for a porcine transmissible gastroenteritis virus gene S and a primer thereof in the technical field of biotechnology. The method comprises the following steps of: cloning a PCR amplification target segment identified as a positive PCR product to a vector pMD18-T, transforming to a competent cell DH5alpha, selecting positive clone by screening blue and white spots and identifying sequencing; extracting a positive recombinant plasmid, quantifying by using an ultraviolet spectrophotometer, diluting a standard product series by 10 times of gradient until the final concentration is 1.0*10<3>-1.0*10<11> copies/mL, undergoing a fluorescence quantitative PCR by taking the standard product series as a template, and establishing a fluorescence quantitative PCR standard curve; and extracting virus RNA (Ribonucleic Acid) of a clinical excrement sample, undergoing a fluorescence quantitative PCR, and calculating the content of viruses in the sample according to a result and the standard curve, wherein the sequences of the primer are sequence 1 and sequence 2. The method and the primer have theadvantages that: a fluorescent probe does not need to be designed additionally, the cost is lowered, operation is easy and convenient, and detection can be completed within 2 hours. The detection method and the primer are suitable for any fluorescence quantitative PCR instrument, and can be applied to the detection of large-scale and high-flux samples.

The invention discloses a quantitative detection method for a gene copy number. The method comprises the following steps of: respectively designing primers and fluorescence probes for target gene and reference gene amplification sequences, and designing a closed probe for the target gene amplification sequence; detecting a sample: performing multiple TaqMan fluorescent quantitative polymerase chain reaction (PCR) on target genes and reference genes by using the DNA sample to be detected as a template, and respectively recording Ct values by using a DNA sample with known target gene copy number as a reference sample; and analyzing the result. By the method, the copy number of the functional genes can be directly and quantitatively detected, and deletion, deletion number and multiple copies (CNVs) of the detected genes are directly reflected, so that synchronous quick detection of gene deletion and multiple gene copies is realized; and the method is high in sensitivity, the synchronous quick detection of the gene deletion and the multiple gene copies can be finished only by a common fluorescent quantitative PCR instrument, and experiments prove that the method is accurate and reliable.

The invention relates to a disc ultra high-speed real-time fluorescence quantitative PCR instrument, comprising a case, three heating blocks which are circularly distributed and have a temperature preset function, a fluorescence detection unit and a matched sealable sampling disc. The circulating amplification of nucleic acid is realized by continuously rotating the disc on different heating blocks for a certain time. According to the invention, the heating and cooling process of the heating blocks in the PCR instrument in the prior art is avoided, and the control program is simplified, so that amplification time of PCR is greatly reduced from more than 1 hour by 5-10 min. The invention can be widely applied in nucleic acid quantitative detection of diseases, especially for rapid detection of paroxysmal infectious diseases to save precious time for treating paroxysmal infectious diseases and controlling the epidemic situation. The instrument has the advantages of fast and convenient operation and high application value.

The invention discloses a kit for jointly detecting breast cancer 21 genes. A PCR (polymerase chain reaction) amplification primer consists of 21 primer pairs, and can be used for jointly detecting 21 gene-related expression quantities: Ki67, STK15, Survivin, CCNB1, MYBL2, MMP11, CTSL2, GRB7, HER2, GSTM1, CD68, BAG1, ER, PGR, BCL2, SCUBE2, ACTB, GAPDH, PRLPO, GUS, and TFRC. Output fragments of the 21 pairs of primers related in the invention are smaller than 100bp, so that PCR efficiency after reverse transcription of mRNA (messenger ribonucleic acid) extracted by a paraffin specimen is greatly improved, all primers of 21 genes are premixed, and time and labor for designing and synthesizing 21 pairs of genes by an experimenter are saved; the corresponding primers of the 21 genes are not needed to be added for premixing in mother liquor, and therefore, errors of reaction holes are greatly reduced. The kit disclosed by the invention is simple to operate, and capable of detecting on a quantitative PCR instrument by only adding a template, so that detection experimental time is greatly shortened. The kit disclosed by the invention is applicable to a single sample and multiple samplers, so that results can be quickly obtained, and therefore, scientific research and clinical detection requirements can be satisfied very well.

The invention discloses a detection method for identifying the American classical PRRSV strain, the HP-PRRSV strain and a new-type viral NADC30 strain at the same time. In the method, a multiplex real-time fluorescence RT-PCR for identifying three kinds of strains at the same time is established based on an ABI7500 fluorescent quantitative PCR instrument, wherein the strains include the classical PRRSV strain, the highly-pathogenic HP-PRRSV strain with the deletion of L-amino acid at the 481st locus of the Nsp2 gene and the continuous deletion of 29 amino acids at the 533ird-561st loci of the Nsp2 gene, and NADC30like; specific primers and specific probes for the three kinds of strains are designed respectively based on the Nsp2 gene, universal primers and universal probes for PRRSV are designed according to the relatively conserved N gene, missing detection caused by Nsp2 region mutation can be effectively prevented, and false negative can be avoided.

The invention discloses a kit for rapidly detecting nucleic acid of a hepatitis C virus (HCV) by a one-step method. The kit comprises a nucleic acid extracting solution, an RT-qPCR reaction solution, a negative control, a positive control and a positive reference. The kit is mainly used for overcoming the disadvantage of the existing kit on the market at present that the time for nucleic acid detection is relatively long. According to the kit, the denaturation of the nucleic acid at a relatively low temperature is achieved through changing ingredients of a reaction buffer and uniquely designing and synthesizing primers and probes on the basis of the conventional RT-qPCR basic reaction principle, then, the temperature difference of denaturation and renaturation during PCR is reduced, and finally, whole reaction time is greatly shortened. The kit disclosed by the invention has relatively high detection sensitivity, specificity and repeatability and can be used for detecting common six genotypes of the hepatitis C virus; meanwhile, the kit is also applicable to the conventional quantitative PCR instrument, so that an accurate basis is provided for auxiliary diagnosis of hepatitis C virus infections and drug therapy monitoring of infected persons.

The invention relates to a novel fluorescent quantitative PCR instrument, and belongs to the technical field of medical equipment. The fluorescent quantitative PCR instrument comprises a kit, an upper plate, elastic rods, springs, a supporting plate, a first gear, a roller, a flange bearing, a controller, a base plate, second motors, supports, shaft sleeves, second gears, second pin rolls, connecting rods, fisheye bearings, connecting posts, first pin rolls, air cylinders, sliding rails, connecting bars, lead screws, sliding blocks, nuts, a bearing support, first motors, motor bases, a PCR instrument main machine and a cover plate. The flange bearing is mounted on the base plate. The roller is mounted on the flange bearing. The first gear is mounted on the roller. The upper end of the roller is mounted to the lower part of the supporting plate. The second gearings are disposed at the right and left sides of the first gear respectively and are meshed with the first gear. The second gears are mounted to the shaft sleeves. The shaft sleeves are mounted on output shafts of the second motors. The fluorescent quantitative PCR instrument is advantageous in that the kit can be automatically opened.

An adjustable fluorogenic quantitative PCR instrument comprises a sample plate, a strut, a rack, a gear motor, a first electric drive pusher, a first electromagnet, a second electromagnet, a first backing plate, support blocks, a liquid crystal display screen, a spring, second backing plates, a fourth electromagnet, a third electromagnet, a second electric drive pusher, a moving block, a linear motor, a PCR instrument mainframe, a controller, lower clamping plates, upper clamping plates, sliding blocks, sliding rails, support rods, a support, a groove, second air cylinders, first air cylinders and a flap, wherein the strut is mounted in the PCR instrument mainframe, the sample plate is placed on the strut, the flap is hinged and mounted on the PCR instrument mainframe, the groove is formed in the flap, the first air cylinders are mounted, the piston rods of the first air cylinders extend out of the groove, the support is mounted on the piston rods of the first air cylinders, the support rods are mounted on the left side and the right side below the support, the second air cylinders are mounted at front ends of the support rods, and the upper clamping plates are mounted below the second air cylinders. The adjustable fluorogenic quantitative PCR instrument has the advantage that the orientation of the liquid crystal display screen can be adjusted.

The invention discloses a multifunctional fluorescent quantitative PCR instrument, and belongs to the technical field of gene detection equipment. The instrument comprises a first electric push rod, a multistage telescoping rod, a first moving block, a first linear motor, a second linear motor, a first clamping plate, a lid, a second gear, a second motor, a belt, a wheel carrier, a first motor, rollers, a support plate, a third sliding block, a third slide rail, a rotation shaft, a cylinder, a first flange bearing, a second slide rail, a platen, a second sliding block, a second flange bearing, a second electric push rod, a first gear, a third gear, a bearing groove, a carrier, a first embossment, a controller, a sample tube, a PCR instrument host, a turnover cover, a second clamping plate, a second moving block, and a casing. The turnover cover is arranged on the PCR instrument host. The wheel carrier is arranged in the PCR instrument host. The left side and right side of the wheel carrier are both provided with a roller. The provided instrument can automatically cover a sample tube.

The inventiondiscloses a kit for directly detecting a folic acid medication sensitive gene by use of blood and an application method thereof. Human blood is adopted as a template, and a real-time fluorescence quantitative PCR system is formed by adding corresponding specific feature primers and fluorescent probes for detecting polymorphism of MTHFR C677T and MTHFR A1298C into the human blood respectively and amplifying a blood resistance taqpolymerase buffer and a blood resistance taqpolymerase. Through a Real time PCR reaction, a person skilled in the art determines the genotype of MTHFR according to the data collected by a fluorescence quantitative PCR instrument. The invention has the advantages of unnecessity of nucleic acid extraction, high speed and time saving, high sensitivity and easy judgment, low cost and the like, and is suitable for clinical detection and promotion.

The invention discloses a detection method of brucella. The detection method comprises the following steps: acquiring brucella DNA from a serum sample by adopting a centrifugal column method, thus obtaining a to-be-detected sample; carrying out PCR amplification reaction; detecting a reaction result by adopting a fluorescence quantitative PCR instrument, wherein during fluorescence signal collection, fluorescein corresponding to fluorophore at the BKV-FP 5' end of a Taqman fluorescence probe is set, and the fluorescence signals are collected at 60 DEG C; and analyzing the result. The invention further provides a kit for detecting the brucella. The detection method and the kit are high in sensitivity, and the minimal detection can achieve 1000 copies/mL. Meanwhile, the detection method is good in specificity, and the cross reaction with other bacteria, such as escherichia coli, salmonella enteritidis, hepatitis B virus, human herpes virus 4, and human cytomegalovirus can be avoided.

The invention discloses a dual-channel real-time fluorescence quantitative PCR instrument light path system and a detection method. The light path system comprises a double-color light excitation module, a micro-fluidic chip module, a dual-channel fluorescence detection module and an image processing module, wherein the micro-fluidic chip module is used for amplifying a to-be-detected sample and collecting fluorescence; the double-color light excitation module is used for enabling the to-be-detected sample to emit fluorescence and separating excitation light emitted by an excitation light source from the fluorescence emitted by the to-be-detected sample; and the dual-channel fluorescence detection module is used for realizing channel transmission of fluorescence with different wavelengthsand adjusting the optical path of the emitted fluorescence with different wavelengths. Fluorescence of the to-be-detected sample is excited through double channels, real-time synchronous imaging of fluorescence of different wavelengths is achieved through a single image capturing device, and the cost of a light path system and the cost of an overall instrument are reduced. The optical path systemis simple in design and compact in structure, optical signal transmission attenuation in the optical path system is reduced, and miniaturization and modularization of the whole instrument are facilitated; interference between exciting light and fluorescence is avoided, and the receiving signal-to-noise ratio of the image capture device and the instrument detection sensitivity are improved.

The invention discloses an isospora suis oocyst detection method based on LAMP (loop-mediated isothermal amplification) real-time fluorescence and an isospora suis oocyst detection primer. A specific primer is designed according to isospora suis 18S rRNA, and together with a fluorescence quantitative PCR instrument, the isospora suis LAMP real-time fluorescence detection technology is built. By adopting the method, the detection limit of an isospora suis recombinant plasmid is 2.74*10<2> copy/[mu]L which is converted into the plasmid DNA concentration of 1 fg/[mu]L; the LAMP detection testing result of a specific number of extracted oocysts shows that the LAMP detection limit is DNA of 5 oocysts. Through building of the LAMP real-time fluorescence detection technology, an extremely valuable detection method is provided for molecular detection of isospora suis, and the shortcomings of a traditional microscope examination method are made up.

The invention provides a fluorescent quantitative PCR instrument based on full-automatic sampling. The instrument comprises a fluorescent detection device, a full-automatic sample processing device and a constant-temperature chamber device, a sample movement guide rail device which can move in the X-axis direction, Y-axis direction and Z-axis direction is arranged between the full-automatic sampleprocessing device and the constant-temperature chamber device, the fluorescent detection device is arranged outside the constant-temperature chamber device, and the full-automatic sample processing device comprises a sample feeding area, a sampling cleaning part and a reaction area which are connected in sequence. The sample movement guide rail device which can move in the X-axis direction, Y-axis direction and Z-axis direction is connected with the full-automatic sample processing device and the constant-temperature chamber device, sample preprocessing of a PCR reaction can be fully automatically achieved, manual operation can be avoided, errors are reduced accordingly, the accuracy is improved, and the detection time of sample amplification is shortened; according to the full-automaticsample processing device, automatic feeding, adding and sampling cleaning of samples can be achieved, large pollution is avoided, and the accuracy of sample detection is further improved.

The invention discloses a real-time fluorescence quantitative PCR (polymerase chain reaction) instrument which comprises basic PCR components, a fluorescence detection component, an upper computer and the like. The real-time fluorescence quantitative PCR instrument has the advantages that PCR portions are foundations of the real-time fluorescence quantitative PCR instrument and include heating modules, heat cover components and temperature regulating modules, basic requirements on accurate temperature control, quick heating and cooling, uniform temperature fields and the like of the real-time fluorescence quantitative PCR instrument can be met by the aid of additional heating of the temperature regulating modules, and smooth PCR procedures can be guaranteed; the fluorescence detection component comprises exciting light systems and detection systems; the upper computer comprise data acquisition and system analysis software and is mainly responsible for acquiring data from lower computers, forming real-time graphs, processing the data and analyzing the graphs to obtain the contents of target DNA (deoxyribonucleic acid) fragments, other detection report information and the like.

The invention relates to a multiplex real-time PCR kit and method for detecting respiratory pathogens and an application. The kit comprises a reagent for detecting influenza A viruses, influenza B viruses, human rhinoviruses and human metapneumoviruses; the reagent comprises a probe for detecting the viruses, a primer pair for specifically amplifying corresponding virus target genes, a reaction buffer solution, an enzyme mixture, a positive control and a negative control; and a detection channel of the influenza A viruses is FAM, a detection channel of the influenza B viruses is VIC, a detection channel of the human rhinoviruses is Texas Red, and a detection channel of the human metapneumoviruses is Cy5. The kit disclosed by the invention can be used for simultaneously detecting the four kinds of viruses, simplifies the operation process, shortens the detection time, improves the detection flux and reduces the detection cost while having high sensitivity and specificity, and can be universally applied to various fluorescent quantitative PCR instruments.

The invention discloses a fluorogenic quantitative PCR instrument. Temperature is controlled with an air heating method, and instrument operation speed is increased greatly. The instrument comprises a fluorogenic quantitative system and a computer, wherein the fluorogenic quantitative system comprises a thermal circulation device, a fluorescence detection device and a controller, the thermal circulation device and the fluorescence detection device are both connected with the controller, and the controller is connected with the computer; the thermal circulation device comprises a temperature controlled heater and a rotary power device, the fluorescence detection device comprises an excitation light path and a detector, and the heating method of the temperature controlled heater is the air heating method.

The invention provides a novel coronavirus detection kit based on Crisper. The crRNA comprises a crRNA targeting a C-terminal part sequence and a crRNA targeting an N-terminal partial sequence, wherein the sequence of the crRNA targeting the C-terminal part sequence is shown as any one of SEQ ID NO.3 and SEQ ID NO.4, and the crRNA targeting the N-terminal partial sequence is shown as any one of SEQ ID NO.5 and 6. The invention alsohe invention relates to the crRNA related to the C-terminal partial sequence and the N-terminal partial sequence of a novel coronavirus. Through application of a method for mutation detection by using the crRNA in combination with a CRISPR-LwaCas13a system, the method has the advantages of low cost, repeated detection for multiple times, simplicity, no dependenceon a special fluorescent quantitative PCR instrument, high detection speed, high sensitivity (the lowest detection limit reaches 10<-2> copies/[mu]L) and strong specificity, and can be used to detectwhether the target nucleic acid is contained or not in a short time through the change of a fluorescence signal.