65 results about "Lysis buffer" patented technology

The invention relates to a method for determining enzyme activity of ODC1 (ornithine decarboxylase 1), which comprises the following steps: digesting, centrifuging and resuspending cells, and taking a part of sample to detect protein concentration; the method comprises the following steps: cracking cells by using a specially-made enzyme activity cell lysis solution and a protein lysis solution, and ensuring full cracking through ultrasonication; carrying out quantitative analysis on the protein in the lysate by adopting a BCA kit; in the enzyme activity detection stage, a cell lysis solution and an enzyme activity detection working solution containing a specific fluorescent probe (CB6) are mixed and react at 37 DEG C for a certain time, and then the change of fluorescence intensity is determined by a multifunctional fluorescence microplate reader, so that the enzyme activity of the ODC1 is evaluated. According to the method, accurate determination of the ODC1 enzyme activity is realized by optimizing cell treatment, protein quantification and enzyme activity detection processes. The method has the advantages of simplicity and convenience in operation, high sensitivity, good specificity and the like, and is suitable for the fields of basic research, drug screening and the like related to the ODC1 enzyme activity.

The invention belongs to the technical field of molecular biology, and particularly relates to a pathogenic microorganism nucleic acid co-extraction kit, an extraction method and application. The invention provides a lysis solution for nucleic acid co-extraction of pathogenic microorganisms. The lysis solution comprises a first guanidine salt, an ionic surfactant, a first sodium salt and a first nonionic surfactant. According to the splitting system, guanidine salt is adopted and matched with a certain content of sodium salt and a surfactant, a sample is split and precipitated under the grinding condition, nucleic acid cannot be damaged in the crushing process, and pathogenic microorganism DNA and RNA with high concentration and complete fragments can be extracted.

The invention discloses a method for extracting cell nucleuses from animal tissues, and relates to the technical field of biology. The background technology indicates that cell nucleus extraction is crucial to multidisciplinary research, but a traditional method has the problems of complex operation, time consumption, low purity, easiness in damaging cell nucleuses and the like, and the prior art is still limited although the prior art is improved. The method comprises the following steps: preparing a sample, and taking fresh or cryopreserved animal tissues; homogenizing the tissues, namely putting the tissues into a homogenizer, and homogenizing at a low speed by adding a cracking buffer solution; carrying out cell lysis, and incubating homogenate on ice; centrifugally separating, centrifuging at low temperature and medium speed, taking precipitate, and discarding supernatant; core washing, re-suspending the precipitate in a washing buffer solution, uniformly mixing and filtering; performing core extraction, re-suspending the precipitate in a suspension buffer solution, and lightly and uniformly mixing; purifying and washing, carrying out density gradient centrifugation on the suspension, dyeing with trypan blue, taking a core layer, and washing for subsequent experiments. The nuclear extraction lysis buffer solution has two proportions of LB1 and LB2, different tissues are suitable for different proportions, and an enzyme inhibitor can be added as required. The method can be used for rapidly extracting various animal tissue cell nucleuses, is complete in shape, large in quantity and few in impurities, is suitable for different tissues, and can be applied to downstream experiments such as single cell nucleus sequencing.

This invention relates to the field of peptide preparation technology, specifically to a preparation process for terfenadine. The terfenadine preparation process includes the following steps: (1) using resin as a solid-phase carrier, Boc-Pip(Fmoc)-OH is loaded onto the resin to obtain Boc-Pip(Fmoc)-resin; (2) under the action of a condensing agent, Fmoc-D-Lys(Boc)-OH and Boc-D-Phe-D-Phe-D-Leu-OH are sequentially coupled onto the Boc-Pip(Fmoc)-resin to obtain terfenadine peptide resin; (3) the terfenadine peptide resin is lysed using a lysis buffer, then concentrated, precipitated, and dried to obtain crude terfenadine. This invention's preparation process can effectively improve product purity while ensuring large-scale industrial production, and the steps are simple with a short reaction cycle.

The present invention relates to a method for isolating and enriching pathogenic nucleic acids from a biological sample thereby reducing host nucleic acid contamination and enrich relative pathogenic DNA in the infectious disease sample and a device thereof. The method involves mixing pathogenic nucleic acid containing biological sample with a lysis buffer at a temperature between 25-35°C for partial or incomplete lysis of the sample; passing the lysed sample through a depth filter wherein the filter paper size ranges from 0.22 microns to 20 microns; passing an elution buffer by reverse plunging action and generating a vibration of 200 to 350 Hz with discontinuous pulse of 20-30 seconds for 2-5 minutes for eluting pathogenic nucleic acids into the elution buffer. The invention further provides a device for isolating and enriching pathogenic nucleic acids from a nucleic acid containing biological sample thereby reducing host nucleic acid contamination using said method.

This invention provides an oligonucleotide cleavage reagent (JS2 & S4 reagent) and its preparation method, as well as a method for cleaving oligonucleotides. The oligonucleotide cleavage reagent comprises NaOH, KOH, methanol, and a cleavage buffer. The oligonucleotide cleavage reagent JS2 & S4 and the cleavage method using it provided by this invention can efficiently cleave oligonucleotides, especially those synthesized using solid-phase phosphorous acid cleavage. This reduces chain breakage and side reactions caused by high-temperature cleavage, and also reduces equipment costs. While shortening the cleavage time, it can effectively reduce oligonucleotide quality problems caused by cleavage.

The invention belongs to the technical field of biomedical detection, and relates to a colon cancer stem cell marker detection kit. The kit comprises a processing assembly, a detection assembly, a reading assembly and a shell, a detection strip is arranged in the detection assembly, and three markers of CD133, LGR5 and ALDH1 are simultaneously detected by adopting a lateral flow immunochromatography technology; the treatment assembly comprises a sample collector, a cracking buffer solution storage cavity and a mixing cavity, and can automatically complete sample pretreatment; the reading assembly comprises an optical detection module and a signal processing module and can quantitatively analyze the detection result. The kit has the advantages of simplicity and convenience in operation, rapidness in detection, high precision, high portability and the like, the detection time is 15-30 minutes, the detection limit can reach the ng / mL level, and the kit is suitable for clinical rapid detection and basic medical institutions and provides technical support for early diagnosis and treatment monitoring of colon cancer.

The application provides a plasmid DNA extraction kit based on an anion exchange column and application thereof, and relates to the technical field of biology.The plasmid DNA extraction kit based on the anion exchange column comprises an anion exchange column, a lysis buffer group, a balance buffer, a washing buffer and an elution buffer; the anion exchange column comprises a solid-phase extraction empty column, upper and lower sieve plates arranged on the inner side of the empty column and located at the bottom of the empty column, and an anion exchange ligand filled between the upper and lower sieve plates; the anion exchange ligand comprises diethylaminoethyl dextran gel DEAE-A50 and diethylaminoethyl cellulose DE52.The application uses domestic fillers and optimizes the ratio of the fillers as the anion exchange ligand, the cost of which is only 1 / 20-1 / 40 of that of imported products, and the operation time and labor intensity are saved through the gravity flow purification process, and the obtained plasmid has high yield, and the transfection effect is equivalent to that of commercially available kits.

The invention belongs to the technical field of animal epidemic disease detection, and particularly relates to an animal epidemic disease high-sensitivity field detection system combining CRISPRCas and digital PCR, a microfluidic digital PCR-CRISPR integrated chip comprises a nucleic acid extraction area and a digital PCR amplification area which are communicated in sequence; the nucleic acid extraction area is pre-coated with magnetic beads and a lysis buffer solution, the digital PCR amplification area is pre-sub-packaged with an animal epidemic disease target specific primer pair, a fluorescent probe and a PCR premix solution, and the CRISPR verification area is pre-coated with crRNA, Cas12a protein and ssDNA fluorescent report probes aiming at a target sequence; the matched detection kit comprises a sample lysis solution, a quality control standard substance, a positive plasmid and a negative control, and the portable fluorescence detection reader can simultaneously read a fluorescence quantitative signal of the digital PCR amplification region and a fluorescence report signal of the CRISPR verification region and automatically output a detection result. The CRISPR verification of the invention further eliminates false positive; reagents are prepackaged in the chip, professional equipment is not needed, and detection is completed within 30 minutes; and at least four animal epidemic disease pathogens can be detected at the same time.

The invention discloses a single cell whole genome methylation and transcriptome biomics library construction method, and relates to the technical field of sequencing methods.The method comprises the steps that a single cell is added into a sorting tube with a preset lysis buffer for lysis, a nucleophilic coating is arranged on the inner wall of the sorting tube to enable cell nuclei to be adsorbed, and supernate containing mRNA is obtained; carrying out reverse transcription on supernate to obtain cDNA (complementary deoxyribonucleic acid) and constructing a transcriptome library; splitting the cell nucleus to release DNA (Deoxyribonucleic Acid) and carrying out hydrosulfite conversion; the transformed DNA is connected with a molecular tag-containing joint to obtain a methylated intermediate product, after exonuclease treatment, the intermediate products with different molecular tags are combined, and the methylated library is obtained through magnetic bead purification, single chain, connection with a sequencing joint at the other end, amplification and purification. According to the technical scheme, RNA and DNA are separated in the same tube and a library is established in parallel, separation and transfer steps are reduced, RNA degradation and nucleic acid loss are reduced, and methylation library establishment flux and biomics data consistency are improved.

This invention discloses a single-cell microarray-based kit and method for in situ quantitative detection of receptor-binding allergen-specific IgE. The kit includes: a single-cell microcavity array chip, an antigen barcode chip, a plastic clamp, IgE standards, fluorescently modified IgE detection antibodies, and auxiliary reagents. During detection, basophil single cells are loaded into the microcavity array chip, lysis buffer is added, and the cells are clamped together with the antigen barcode chip. Low-temperature incubation allows the allergen-specific IgE released from cell lysis to be captured in situ by the antigen on the chip. A sandwich immunoassay is then performed using a fluorescently labeled antibody, and absolute quantification is achieved using a standard curve. This invention is the first to achieve high-throughput, high-sensitivity, multi-component parallel in situ quantitative detection of basophil membrane receptor-binding sIgE at the single-cell level. It can directly reflect the functional sensitization state of effector cells and has advantages such as simple operation, low sample requirement, and no need for complex valve devices.

This invention provides a lysis buffer for nucleic acid extraction and its application. The lysis buffer comprises: 0.6M–0.8M NaH₂PO₄, 0.042–0.06M Na₂HPO₄, 0.007–0.009M NaCl, 0.03–0.05M Tris, 0.01–0.02M SDS, and 10%–20% anhydrous ethanol; wherein the pH of the lysis buffer is 5–6; and the nucleic acid is DNA and / or RNA. Using the lysis buffer of this invention, the sample can be efficiently lysed, effectively releasing and protecting the nucleic acid molecules in the sample, significantly improving the nucleic acid extraction efficiency. This invention provides a new reference for extracting DNA and / or RNA from low-biomass samples, contributing to improved sequencing quality of low-biomass samples.

The invention discloses an injector-based nucleic acid purification device, an extraction method and a viral nucleic acid lysate. The purification device comprises an injector assembly and a filter membrane assembly, wherein the injector assembly comprises an injector tube barrel and a push rod matched with the injector tube barrel; the filter membrane assembly is connected to the outlet end of the injector tube barrel, and the filter membrane assembly is configured to be of a detachable structure. The nucleic acid purification device provided by the invention perfects the processes of nucleic acid release, nucleic acid washing and elution, can remove components which interfere with subsequent PCR (Polymerase Chain Reaction), fluorescent quantitative PCR and constant-temperature detection in the nucleic acid release process, and can improve the detection efficiency; the viral nucleic acid lysate is suitable for common clinical samples such as nasal swabs, anus swabs, serum and ascites, and is suitable for nucleic acid extraction and purification of DNA and RNA viruses by optimizing the proportion of guanidine isothiocyanate, polyvinylpyrrolidone (PVP-40) and lauryl sodium sulfate (SDS).

This invention relates to the field of nucleic acid detection technology, specifically disclosing a liquid-phase nucleic acid chip for isothermal amplification of common algal blooms and its application. The method includes: collecting water samples and filtering to enrich algal cells; lysing the algal cells using cell lysis buffer to extract DNA; adding the extracted DNA to the liquid-phase nucleic acid chip for isothermal amplification; detecting the Ct value using a qPCR instrument; and determining the species and cell density of the algal bloom based on a preset standard curve. This invention directly quantifies algal cell density using algal gene copy number, with significantly higher accuracy than microscopic counting, and does not rely on the experience of professional personnel. It can be widely applied in frontline water resource management work.

This invention relates to the field of fecal genomics kit technology, and particularly to a safe, rapid, and high-throughput kit for extracting fecal genomes and its usage method. The kit includes a sample preservation solution, magnetic beads, a lysis buffer, a washing buffer 1, a washing buffer 2, and an elution buffer. The sample preservation solution is composed of guanidine isothiocyanate, sodium citrate, Triton X-100, Tween 20, propidium iodide, and purified water. The magnetic beads are hydroxyl magnetic beads. The lysis buffer is composed of guanidine hydrochloride, Tris base, Tris hydrochloric acid, Triton X-100, isopropanol, and purified water. Washing buffer 1 is composed of guanidine hydrochloride solution and isopropanol. Washing buffer 2 is composed of anhydrous ethanol and purified water. This kit can easily extract the genome from fecal samples, is simple to operate, safe and non-toxic, and can extract multiple samples at once, enabling rapid diagnosis and filling the gap in the current market for automated high-throughput kits for extracting genomes from fecal samples.

This invention discloses a method for protein extraction and mass spectrometry pretreatment from methanogenic bacteria, belonging to the field of protein extraction technology. The method involves pretreating a methanogenic bacteria enrichment culture to remove inorganic salt ions and other impurities, resulting in a pretreated culture. Then, a lysis buffer composed of sodium dodecyl sulfate, Tris-HCl buffer, and a protease inhibitor is added to the pretreated culture, followed by ultrasonic disruption to obtain a cell lysate, from which a crude protein extract solution is obtained. Finally, the protein extract is extracted from the crude protein extract solution. This invention significantly improves protein extraction efficiency and peptide purity by optimizing the extraction process and subsequent enzymatic digestion, while preserving the methanogenic activity of the extracted protein as much as possible, providing more accurate and reliable sample support for downstream protein mass spectrometry analysis.

This invention belongs to the field of biotechnology and discloses a plasma cell-free DNA extraction kit, extraction method, and applications. The kit includes magnetic beads, lysis buffer, proteinase K, binding buffer, washing buffer, and elution buffer. The extraction method involves mixing the plasma sample, lysis buffer, proteinase K, binding buffer, and magnetic beads. The lysed plasma cell-free DNA (cfDNA) is incubated with the magnetic beads to form a magnetic bead-cfDNA complex. Under a magnetic field, the complex is washed to remove various impurities, and the plasma cfDNA is obtained after elution. The method of this invention is simple and rapid, with short sample lysis time and stable quality. Replacing isopropanol with anhydrous ethanol during lysis results in high nucleic acid recovery, making it suitable for plasma cfDNA extraction. The purity is suitable for subsequent sequencing and PCR / Q-PCR. The magnetic bead method is beneficial for large-scale use and has significant implications for research and clinical applications.

The invention discloses a method for high-throughput extraction of DNA from cotton, and belongs to the technical field of plant biology. The invention provides a lysis buffer solution for high-flux extraction of DNA (deoxyribonucleic acid) from cotton, which comprises the following components: 50-80 g / L of NaOH, 2-4 g / L of NaCl, 0.1 g / L of SDS (sodium dodecyl sulfate), 0.8-1.5 g / L of EDTA-2Na. 2H2O, 10-30 g / L of PVP (polyvinyl pyrrolidone), 2 g / L of gelatin and 8-12 g / L of ascorbic acid, and all the components are dissolved in deionized water. According to the high-throughput DNA extraction method for the cotton tissues, efficient DNA extraction of the cotton tissues can be realized, the extraction steps are greatly simplified, high-throughput extraction can be carried out by a single person in a laboratory, DNA for PCR amplification is efficiently obtained, reliable sample support is provided for subsequent molecular biology research, and a convenient tool is provided for researchers.

The invention discloses a tissue cell nucleus extraction method and a cell nucleus protective agent applied to flow sorting, and relates to the technical field of cell extraction.The tissue cell nucleus extraction method comprises the steps that operation instruments needed by an experiment are prepared and pre-cooled; cutting a fresh tissue sample into tissue blocks, and immediately transferring the tissue blocks into a precooled lysis buffer solution; manually grinding the lysis buffer solution containing the tissue block by using a homogenizer, and sampling and observing homogenate in the grinding process; filtering the homogenate product through a cell filter screen in sequence; performing low-speed centrifugation on the filtrate by adopting a centrifugal machine, and discarding supernatant to obtain primary cell nucleus precipitate; re-suspending the primary cell nucleus precipitate by using a cell nucleus washing buffer solution to obtain a purified cell nucleus precipitate; and precipitating and resuspending the purified cell nucleuses, counting by trypan blue dyeing, and calculating the percentage of the complete cell nucleuses. According to the method, the problem of more cell nucleus breakage in the tissue cell nucleus extraction process is solved through a mild homogenization strategy monitored under the microscope.

The invention discloses colloidal gold chromatography test paper and a kit for detecting platelet CD36 antigen, the colloidal gold chromatography test paper comprises a bottom plate, a sample pad, a gold-labeled combination pad, a nitrocellulose membrane and a water absorption pad, the gold-labeled combination pad is coated with a colloidal gold labeled mouse anti-human CD36 IgG2a monoclonal antibody and a colloidal gold labeled quality control rabbit IgG polyclonal antibody; the nitrocellulose membrane is provided with a detection line coated with a human platelet CD36 antigen capture antibody and a quality control line coated with quality control goat anti-rabbit polyclonal antibody molecules. When the kit is used for detecting the CD36 antigen, a detected sample is fully split by the platelet lysis buffer and then is added into the sample adding hole of the kit, and a detection result can be observed after waiting for 10-15 minutes. By adopting the detection mode provided by the invention, the CD36 antigen on the human platelets can be quickly, simply, conveniently, sensitively and specifically detected.

The invention belongs to the field of quantitative analysis of drug concentration of a cell lysis solution, and particularly discloses a method for detecting estradiol 17 beta-D glucuronic acid (Estradil 17-(beta-D-Glucuronide)) in the cell lysis solution. According to the method, the concentration of the estradiol 17 beta-D glucuronic acid in the cell lysis buffer can be accurately quantified through specific sample treatment and liquid chromatography and mass spectrometry conditions, the sensitivity is high, the detection limit is low, a novel method capable of being practically popularized and applied is provided for pharmacokinetic research of the estradiol 17 beta-D glucuronic acid, and the method has popularization and application values.

This invention discloses a method for detecting exosomes, including the step of mixing a biological sample with a lysis buffer to obtain a sample lysis buffer; the volume ratio of the lysis buffer to the biological sample is 1:1. Using both exosome extraction and direct lysis methods of this invention to detect the concentrations of TSG101 and PDCD6IP, the detection results show high consistency, indicating that this method improves efficiency while maintaining high accuracy.

The present invention relates to a cell lysate wherein the lysis buffer comprises magnesium acetate, the phosphorylation inhibitor K3L, the agonist of dephosphorylation GADD34 and a caspase inhibitor for use in, or as, a cell-free system and a method for preparing a cell-free protein synthesis system. Further provided is a method for expressing a protein in the cell-free protein synthesis system, a method for screening for a candidate compound for an effect on a target molecule and a use of the cell-free protein synthesis system. The present invention also provides a kit for the above-mentioned cell-free systems.

This invention provides a lysis buffer, kit, and method for extracting microbial nucleic acids. The lysis buffer contains guanidine salt, anionic surfactant, inorganic salt, nonionic surfactant, tris(hydroxymethyl)aminomethane, and phosphite. By using phosphite in combination with other components in the lysis buffer, the lysis effect on structurally complex microorganisms such as fungi is improved without affecting the nucleic acids of structurally simple microorganisms such as viruses released earlier. This invention also provides a nucleic acid extraction kit containing the lysis buffer, enabling the co-extraction of nucleic acids from multiple microorganisms. The kit and extraction procedure of this invention, when used together, can obtain high-quality nucleic acids from different microorganisms in various infected samples; it shortens the extraction time for different microbial nucleic acids, simplifies the operation process, and reduces extraction costs.

The invention discloses a preparation method and application of bionic nano immunomodulatory bait, and the preparation method comprises the following steps: firstly, obtaining neutrophils, treating the neutrophils with a cell lysis buffer solution containing a calcium ion chelating agent and a protease inhibitor, collecting supernate, and then centrifuging and purifying to obtain neutrophils membranes; then mixing an organic phase containing a polymer polylactic acid-glycolic acid copolymer and a therapeutic drug with a polyvinyl alcohol solution, and emulsifying to obtain a PLGA-therapeutic drug nano core; and finally, co-extruding the material and the neutrophile granulocyte membrane. The neutrophile granulocyte membrane is used as a camouflage shell, the nano bait capable of efficiently targeting an inflammatory site and neutralizing multiple inflammatory mediators is constructed, the nano bait is applied to brain injury after cardio-pulmonary resuscitation, efficient and active targeting on an inflammatory brain area can be achieved, the death rate of neurons can be reduced, and the nano bait has an extremely good application prospect.

The invention relates to the technical field of cell biomedicine, in particular to umbilical cord mesenchymal stem cells with high PD-L1 expression, a culture method and application of the umbilical cord mesenchymal stem cells and a method for promoting PD-L1 expression through in-vitro energization. The umbilical cord mesenchymal stem cell in-vitro energizing inducer is based on a DMEM / F12 culture medium containing 3%-7% of human platelet lysis buffer, and further comprises the following components: a tumor necrosis factor alpha, interferon gamma, human serum albumin, transferrin, insulin, riboflavin, N-acetylcysteine and a vascular endothelial growth factor. The in-vitro energizing inducer reduces the content of a stimulating factor tumor necrosis factor alpha and interferon gamma, reduces immunogenicity abnormity caused by TNF-alpha, remarkably improves PDL1 expression and provides a basis for inflammation treatment. The influence of long-time stimulation on cell functions and activity is remarkably reduced through short-time factor-induced stimulation.

This invention discloses a method for determining polyamines in breast milk and dairy products by gas chromatography-tandem mass spectrometry (GC-MS / MS), comprising: taking 20–200 μL of breast milk or dairy products, adding an isotope mixed standard solution, mixing well, then adding lysis buffer, vortexing thoroughly, centrifuging, taking the supernatant, centrifuging to concentrate and drying, resuspending the dried sample in a mixture of ethyl acetate and trifluoroacetic anhydride, then derivatizing in a water bath at 40–80 °C for 0.5–3 hours, continuing centrifugation to concentrate and dry after derivatization, redissolving in dichloromethane, performing solid-phase extraction purification, blowing the purified solution to near dryness with nitrogen, adding 20–200 μL of a mixture of isopropanol and ethyl acetate to redissolve, and performing GC-MS / MS detection; qualitatively identifying the polyamines based on the qualitative ion pairs and retention times; substituting the ratio of the peak area of ​​the quantitative ion of the polyamine to the peak area of ​​the quantitative ion of the deuterated isotope internal standard into the matrix-matched internal standard curve of the polyamine to obtain the concentration of the polyamine.

The invention provides a polar charged residue engineered MspA nanopore, the MspA nanopore is based on a mutant M2 MspA nanopore, site-specific mutagenesis is carried out on an N91 amino acid site in a contraction region of the MspA nanopore, and mutated amino acids comprise aspartic acid, arginine and glutamic acid. The preparation method of the nanopore comprises the following steps: S1, inserting a gene of a mutant M2 MspA subjected to site-specific mutagenesis into a plasmid; s2, transforming the recombinant plasmid into an escherichia coli competent cell, culturing, cooling a culture system, adding IPTG (isopropyl-beta-d-thiogalactoside), continuously culturing, and inducing the expression of a target protein; and S3, collecting the thalli, adding a cracking buffer solution, heating, cooling, centrifuging, collecting supernatant, and purifying by nickel affinity chromatography. The MspA nanopore provided by the invention can realize single molecule detection of 10 kinds of PTMs and can also distinguish isomers, the recognition accuracy rate exceeds 98%, and a novel tool is provided for complex modification proteomics research.

This invention discloses a disc-shaped microfluidic chip detection kit and method, relating to the field of rapid food safety detection. A disc-shaped substrate has a ring-shaped central flow channel, and in the direction of rotation towards the central flow channel, a lysis buffer chamber, an elution buffer chamber, a first washing buffer chamber, a second washing buffer chamber, and a magnetic bead reagent chamber are arranged in a ring-shaped sequence. At least four reaction chambers are arranged in a ring-shaped interval in the direction away from the rotation axis of the central flow channel. A ring magnet is positioned on the bottom plate facing the central flow channel, and a pin for opening a contact valve is arranged around the periphery of the ring magnet. Two types of heating plates with temperature differences are respectively positioned on the heating plate facing the reaction chambers. This invention relies on centrifugal force to achieve autonomous fluid drive, combined with constant temperature zone control to complete rapid amplification. It not only eliminates cross-contamination problems caused by magnetic bead transport at the structural level, but also achieves a high degree of integration of the entire process from nucleic acid extraction to fluorescence amplification, effectively improving the overall detection efficiency of pathogens.