57 results about "Reference gene" patented technology

The invention provides a method and a device for designing a protein coding codon sequence based on a large multi-modal model, which comprehensively utilize information such as an amino acid sequence, space folding and mRNA quality of protein to realize high-quality, controllable and evaluable codon sequence design. According to the method, the dependence of a traditional codon optimization strategy on a reference gene template is broken through, and a codon optimization problem is converted into a codon design task based on protein information. Through comprehensive consideration of information such as space folding of protein, context of amino acid sequences, mRNA quality and the like, the model can generate a codon sequence which is more in line with codon use preference of specific species, reasonable in structure and efficient in expression, so that a new thought and technical support are provided for safe, stable, efficient and controllable protein expression.

The invention provides a primer probe combination and a kit for detecting the copy number of a 22q11 region based on digital PCR and a use method of the primer probe combination. Specific probes of a target gene TBX1, a target gene CRKL, a target gene SMARCB1 and a reference gene GAPDH are integrated in different fluorescence channels, and the copy number of the target gene is detected according to the ratio of the copy number of the reference gene to the copy number of the three target genes. According to the invention, amplification products of different sites in different fluorescence channels are integrated in the same system in digital PCR reaction, so that the detection flux is greatly improved, and low-cost and high-flexibility accurate quantification of 22q11. 2 copy number variation is realized through a single-channel multi-target fluorescence layering strategy; and the dependence on expensive equipment, complex operation and professional analysis in the prior art is overcome.

The invention discloses a primer probe combination and a kit for on-site detection of a transgenic maize transformant, and relates to the technical field of transgenic detection. The invention provides a primer probe combination for on-site detection of a transgenic maize transformant. The primer probe combination comprises a primer pair and a probe for detecting a transgenic maize strain MON810, the primer pair and the probe are used for detecting the transgenic maize strain TC1507; the primer pair and the probe are used for detecting the transgenic maize strain Bt11; the invention discloses a primer pair and a probe for detecting corn internal reference zSSIIb. The method for detecting the transgenic maize by adopting the primer probe combination and the kit disclosed by the invention integrates sample pretreatment, nucleic acid extraction, purification, amplification and detection, has better sensitivity and accuracy, is simple, convenient, rapid and time-saving to operate, and plays an important role in on-site rapid detection of transgenic maize research and safety management.

The invention relates to the technical field of molecular biology and environmental biology, and discloses a PCR (Polymerase Chain Reaction) chip and a kit for detecting the expression quantity of functional genes related to microbial degradation of chlorinated hydrocarbon and application of the PCR chip and the kit. The PCR chip comprises a primer pair and a primer pair, the primer pair is used for specifically amplifying dehalogenation functional genes bvcA, cfrA, mbrA, pceA, pteA, tceA and vcrA respectively, the primer pair is used for specifically amplifying reference genes, and nucleotide sequences of the primer pair for specifically amplifying the dehalogenation functional genes are shown as SEQ ID NO. 1 to SEQ ID NO. 14. The PCR chip provided by the invention can specifically amplify seven functional genes related to the degradation of the chlorinated hydrocarbon microorganisms, is good in specificity, high in accuracy and good in repeatability, and can accurately detect the expression condition of the seven dehalogenation functional genes related to the degradation of the chlorinated hydrocarbon microorganisms through a qPCR technology, so that the existence condition of chlorinated hydrocarbon degrading bacteria can be reflected; the method has reliability in the aspects of chlorohydrocarbon polluted site biodegradability evaluation, degradation process research, repair effect verification and the like.

The invention discloses a reference gene for acer ginnala gene expression analysis as well as a primer and application thereof, and belongs to the technical field of plant molecular biology. The reference gene is AgTUA and / or AgPP2A and / or AgACT2, the nucleotide sequence of the AgTUA gene is as shown in SEQ ID NO.4, the nucleotide sequence of the AgPP2A gene is as shown in SEQ ID NO.6, and the nucleotide sequence of the AgACT2 gene is as shown in SEQ ID NO.2. The invention further discloses a kit for detecting the content of the AgPP2A gene and the AgACT2. The invention also provides amplification primers for detecting the reference gene, and the primer sequences of the amplification primers are respectively shown as SEQ ID NO.16 and SEQ ID NO.17, SEQ ID NO.20 and SEQ ID NO.21, and SEQ ID NO.12 and SEQ ID NO.13. The invention also provides a kit for detecting the reference gene. The reference gene is suitable for real-time fluorescent quantitative expression analysis of acer ginnala flavonoid synthesis related genes or other functional genes, the current situation that acer ginnala has no reference gene is filled, and the accuracy and reliability of gene expression analysis data are improved; powerful support is provided for acer ginnala functional gene expression analysis and regulation mechanism research.

The invention provides an NOD / shiLt mouse genotype fluorescent quantitative identification method and application, and relates to the field of experimental animal genetic quality control. Comprising the following steps: acquiring a to-be-detected mouse genetic identification sample, and performing DNA extraction treatment on the mouse genetic identification sample to form a genome DNA template; according to the fluorescent quantitative identification method for the genotype of the NOD / shiLt mouse and the application of the fluorescent quantitative identification method, a real-time fluorescent quantitative polymerase chain reaction system containing a specific primer pair, an insertable fluorescent dye premixed solution and a standard substance series is constructed, so that accurate quantitative detection on a specific deletion sequence of a No.3 chromosome of the NOD / shiLt mouse and a reference gene GAPDH is realized. According to the present invention, the linear relationship between the Ct value and the template copy number is established through the standard substance series, and the quality determination of the amplification curve, the melting curve and the reference gene is combined, such that the NOD / shiLt mouse genotype fluorescence quantitative identification method can stably and rapidly identify whether the mouse genomic DNA contains the NOD / shiLt mouse genotype.

The invention belongs to the technical field of virus detection, and particularly relates to a double-channel real-time fluorescent quantitative PCR (Polymerase Chain Reaction) detection primer, a probe, a detection kit, a detection method and application thereof. The detection sensitivity is high, and the lowest detection limit can reach 10 copies / mL; the detection time period is short, nucleic acid detection can be completed within 45 minutes, and the kit is suitable for clinical and bedside rapid detection and diagnosis; the detection specificity is high, and cross amplification with other 80 pathogens is avoided; the detection accuracy is 100%; virus quantitative analysis can be carried out, and the quantitative linear range is good; the detection repeatability is good, and the precision is high; uDG enzyme is added into a detection system, so that pollution can be prevented; a reference gene is also added into the detection system, so that the quality of the whole extraction and amplification process of the sample can be monitored according to the detection result of the reference gene, and the existence of the RNA of the Enduma virus in samples such as whole blood, serum, plasma, cerebrospinal fluid, urine, alveolar lavage fluid, throat swab and excrement can be detected.

The invention relates to a method for inferring a silkworm chrysalis stage development process based on trimethyl lysine hydroxylase. The method comprises the following steps: (1) constructing a correlation model of relative transcription expression quantity of a trimethyl lysine hydroxylase gene and a reference gene of a silkworm chrysalis sample of a variety to be detected and pupal stage development time; and (2) determining the relative transcription expression quantity of the trimethyl lysine hydroxylase and the reference gene in the silkworm chrysalis to be detected, and calculating the pupal stage development time according to the correlation model obtained in the step (1). According to the method, the development process of one or more silkworm chrysalis can be accurately measured, errors caused by traditional visual inspection can be reduced, a new detection method is provided for deducing the pupal development process of the silkworms, and an important basis is provided for judging moth formation time in silkworm seed production work.

The invention relates to the technical field of nucleic acid detection, in particular to an endogenous reference gene, a probe primer and a kit for quantitatively detecting HCMV nucleic acid, the endogenous reference gene comprises a core sequence and flanking sequences located at the two ends of the core sequence, and the nucleotide sequence of the core sequence is shown as SEQ ID NO.1. According to the endogenous reference gene sequence, the detection primer and the detection probe of the endogenous reference gene and the detection primer and the detection probe for specifically recognizing the HCMV nucleic acid, rapid and accurate quantitative detection of the HCMV nucleic acid can be realized, and the accuracy and the reliability of a detection result are improved.

The present invention provides a method for detecting the insertion copy number of foreign genes in Setaria viridis or foxtail millet by fluorescence quantitative PCR or digital PCR, belonging to the field of biotechnology. The method uses RA1 gene as an internal reference gene and Hyg screening marker gene as a foreign gene, and uses primer pairs RA1-F / RA1-R and primer pairs Hyg-F / Hyg-R to detect the genomic DNA of transgenic Setaria viridis or transgenic foxtail millet, and determine the insertion copy number of foreign genes in transgenic Setaria viridis or transgenic foxtail millet. The present invention provides a method for detecting the insertion copy number of foreign genes in Setaria viridis or foxtail millet by fluorescence quantitative PCR or digital PCR, which has the characteristics of simplicity, rapidity, low sample consumption, high efficiency, high throughput, etc., can improve the efficiency of screening transgenic lines of Setaria viridis or foxtail millet with low-copy insertions, and provides a new option for the detection method of the insertion copy number of foreign genes in the research of transgenic Setaria viridis or transgenic foxtail millet.

The invention provides a reference gene combination for real-time fluorescent quantitative PCR (Polymerase Chain Reaction) of indocalamus as well as a primer pair combination and application of the reference gene combination, and belongs to the technical field of plant molecular biology. In combination with indocalamus transcriptome data analysis and cross validation of four algorithms, reference genes, MD10B, PP2A, eIF1A, Ite23725, eIF4A, 60S and UBP1, suitable for indocalamus are systematically screened out, optimal reference gene combinations under different experimental conditions (drought, salt, waterlogging stress and different tissues) are refined, and the method is scientific and rigorous and has good application prospects. And a precise tool is provided for gene expression analysis of the indocalamus in different research directions. The screened reference genes show extremely high expression stability in specific stress and tissues after being combined, and the expression stability is far better than that of traditional housekeeping genes such as Actin and UBI. The specific primer has high amplification efficiency, and ensures the accuracy of qRT-PCR results.

The application belongs to the technical field of molecular diagnosis, and particularly relates to a digital PCR kit and method for accurately detecting the copy number of a CAR gene of a CAR-T cell and a lentivirus vector. The application first realizes the absolute quantification of CAR, Rev and an internal reference gene RNase P in a single tube simultaneously, and has strong universality. Based on the principle of digital PCR, the absolute copy number is directly obtained without a standard curve, and the result is accurate and highly repeatable. The detection lower limit reaches 0.01%, and extremely low abundance CAR-T residues or microresidual diseases can be detected. Primers and probes are designed for the conservative regions of CD3zeta, Rev and RNase P, and no cross reaction is verified, so that false positives are effectively avoided. The sample only needs gDNA, and there is no requirement for cell activity, and the sample is compatible with freezing and transportation. The kit is premixed and optimized, the process is automated, and 4-5 hours are needed. The kit is suitable for the whole chain of CAR-T drug research and development, production process quality control, clinical patient monitoring and safety evaluation.

The present invention discloses a molecular marker combination for detecting adulterated Bos genus milk in buffalo milk and its application, including molecular marker 1 and molecular marker 2; the molecular marker 1 is located at the 2552nd base of the buffalo genus reference gene sequence NC_049568 or the 2540th base of the Bos genus reference gene sequence AB074968, and the polymorphism is T or C; the molecular marker 2 is located at the 2555th base of the buffalo genus reference gene sequence NC_049568 or the 2543rd base of the Bos genus reference gene sequence AB074968, and the polymorphism is G or A. The molecular marker combination of the present invention can be effectively used for detecting adulterated Bos genus milk in buffalo milk.

The invention discloses a noninvasive exosome mRNA BRAF V600E mutation Tag-PCR (Tag-Polymerase Chain Reaction) detection method, which comprises the following steps: separating an exosome sample from a body fluid; the method comprises the following steps: by taking mRNA as a template, adding an exogenous reference gene and a primer pair for detecting the exogenous reference gene, and meanwhile, carrying out a first round of PCR amplification reaction by using a Tag-ARMS primer pair for specifically enriching exosome mRNA BRAF gene V600E mutation so as to form a PCR product; a PCR product is used as a template, a system without UNG enzyme is used for amplification, a primer pair for detecting a reference gene is added, a Tag primer pair for specifically enriching and detecting V600E mutation of the human exosome mRNA BRAF gene and a probe sequence are used at the same time, and then a second round of PCR amplification reaction is carried out; and judging the mutation state of the V600E site of the BRAF gene of the mRNA of the exosome sample.

The invention discloses a brassica chinensis nitrate stress reference gene and primers and application thereof, and relates to the field of molecular biology, the reference gene is a Bc60S RPL4-1 gene, the nucleotide sequence of the Bc60S RPL4-1 gene is shown as SEQ ID NO: 2, and the nucleotide sequence of the Bc60S RPL4-1 gene is shown as SEQ ID NO: 3. A primer for amplifying the reference gene comprises a forward primer with a nucleotide sequence as shown in SEQ ID NO. 10 and a reverse primer with a nucleotide sequence as shown in SEQ ID NO. 11. The invention provides a reference gene Bc60S RPL4-1 gene which is highly stable in expression in brassica chinensis nitrate stress research, and the accuracy and reliability of quantitative analysis of brassica chinensis nitrate stress related gene expression are remarkably improved.

The application discloses a kit for detecting LINC01480 gene expression and a preparation method and application thereof, relates to the field of gene expression kits, and is used for detecting LINC01480 gene expression and comprises a specific primer pair for the LINC01480 gene, a specific primer pair for a reference gene GAPDH, a positive control lentivirus vector containing a LINC01480 gene segment, a negative control lentivirus vector and an RT-qPCR reaction system. The application non-invasively monitors MRD of MM patients after treatment by detecting LINC01480 gene expression, so as to improve the convenience and accuracy of index monitoring of MRD of the MM patients after treatment.

The invention discloses a composition for detecting spinal muscular atrophy typing based on nucleic acid mass spectrometry and application of the composition. The invention relates to the field of gene detection, and provides a composition for detecting spinal muscular atrophy typing based on nucleic acid mass spectrometry, which comprises a target gene primer pair, a reference gene primer pair and an IIS type restriction enzyme. The composition can be used for detecting spinal muscular atrophy typing based on nucleic acid mass spectrometry, and the traditional technical method of nucleic acid mass spectrometry is improved, so that nucleic acid mass spectrometry detection does not need the steps of SAP treatment and single base extension reaction, the reaction time is shortened, the result is accurate and reliable, the steps are simple, and the cost is greatly reduced. The invention is of great significance to preparation of screening products suitable for large-scale screening of spinal muscular atrophy.

The invention relates to the technical field of kits, and discloses a peanut allergen component fluorescence PCR (Polymerase Chain Reaction) detection equivalent kit, which comprises (1) 2 * Probe qPCR Mix; (2) a florescence source probe primer mixture; (3) a 18S internal reference probe primer mixture; (4) ultrapure water; (5) positive control; (6) negative control; according to the invention, a dual detection mechanism of a florescence source specific gene and an eukaryote general reference gene is combined, and a specially designed chimeric recombinant plasmid is adopted as a multifunctional positive control, so that when a complex food matrix is treated, a forced standard reaching mechanism of a reference gene channel can timely identify a sample treatment abnormal condition; and wrong interpretation caused by improper operation is avoided. Meanwhile, the activity of core components of the kit can be verified through the two-channel synchronous response characteristic of positive control, and the credibility of a detection result and the scientific preciseness of interpretation are improved.

This invention discloses a one-step RT-PCR method for detecting five viruses in *Calamus sylvestris*, belonging to the field of plant virus molecular detection technology. Specifically, it provides a primer set for the simultaneous detection of multiple viruses in *Calamus sylvestris*, including five pairs of virus-specific primers for the specific detection of PVBV, CaYMAV, CaYMV, CaYSV, and SCMV, and one pair of *Calamus sylvestris* internal reference gene primers, the sequences of which are shown in SEQ ID NO:1-12. RNA is extracted from *Calamus sylvestris* samples, a one-step RT-PCR reaction system is prepared, amplification and electrophoresis are performed, the detection validity is verified by the internal reference gene band, and the type of virus infection is determined by the virus-specific bands. This invention can achieve simultaneous detection of five viruses and has the advantages of simple operation, short detection cycle, high sensitivity, and strong specificity, making it suitable for rapid screening of large batches of *Calamus sylvestris* samples.

The invention belongs to the field of gene detection, and discloses a method for detecting peripheral plasma of a pregnant woman through ddPCR to screen a fetal DGS syndrome. A plurality of pairs of 22q11. 2 region specific primers are meticulously designed, and the annealing temperature of the primers is controlled to be 62 + / -3 DEG C, so that high consistency of microdroplet amplification efficiency and high quantitative accuracy are effectively ensured, and the defect that gene deletion detection is difficult to realize by ddPCR is overcome. According to the method of some examples, a target region causing the 22q11.2 deletion syndrome and conserved regions of other chromosomes are detected at the same time through droplet type digital PCR, and whether fragment deletion exists in the 22q11.2 region of the fetal chromosome in peripheral plasma of a pregnant woman or not is judged by analyzing the copy number and proportion of a target gene and a reference gene. Therefore, antenatal screening or antenatal auxiliary diagnosis is carried out on whether a fetus suffers from the 22q11. 2 deletion syndrome or not, and the operation is simple and economical.

The invention discloses a digital PCR detection system, kit and method for detecting single nucleotide polymorphism and copy number variation of a CYP2D6 gene and application, and belongs to the technical field of molecular biology. The method comprises the following steps: designing three pairs of CYP2D6 and reference gene RPP30 specific primers, matching with four locked nucleic acid modified TaqMan fluorescent probes, constructing a single-tube multiple digital PCR detection system, and synchronously and accurately detecting three single nucleotide polymorphisms of CYP2D6 * 10, CYP2D6 * 36 and CYP2D6 * 41 and CYP2D6 * 5 copy number variation; wherein the RPP30 reference gene can synchronously realize sample amplification quality control and target gene absolute quantitative calibration. The detection system depends on the advantages of high specificity of the LNA probe and high sensitivity of digital PCR, the detection sensitivity reaches 0.02 ng / mu L, the detection period is short, and the result is stable and reliable.

The invention belongs to the technical field of biological agriculture, and particularly relates to an InDel molecular marker of brassica juncea, a primer combination, a kit and application of the InDel molecular marker. The InDel molecular marker of the leaf mustard is located at the position 30727008-30728104 of an A06 chromosome, and the green leaf sequence of the InDel molecular marker is as shown in SEQ ID NO. 1; the sequence of the silver leaf is as shown in SEQ ID NO.2, and the reference gene is Brassica juncea cv.tumidaT84-66V1.5. The invention also discloses a method for preparing the Brassica juncea cv.tumidaT84-66V1.5 gene. The leaf character of a plant containing the InDel molecular marker as shown in SEQ ID NO.2 is silver, otherwise, the leaf character is green. The invention discloses a primer combination, a kit containing the primer combination and a method for detecting leaf color of mustard. The sequences of upstream and downstream primers are shown as SEQ ID NO.25-26. The method is of great significance to early prediction, screening or breeding of leaf color of leaf mustard.

The invention relates to a real-time quantitative polymerase chain reaction (qPCR) method and a kit, which are used for simultaneously determining classification of specific gene variation and gene copy number. More specifically, the present invention relates to an efficient qPCR method and kit for simultaneously determining the variation and copy number of a target gene by performing real-time quantitative PCR on a reference gene and a target gene with known copy number in the same reaction vessel.

The present invention discloses a kind of reference gene and construction method of real-time fluorescence quantitative PCR for analyzing gene expression under heat stress of indica rice, and belongs to the field of plant genetic engineering technology. The nucleotide sequences of 3 reference genes of the present invention are shown in sequence table SEQ ID NO.1-10;The flag leaf of II excellent 838 and its parent after high temperature stress is subjected to transcriptome high-throughput sequencing;Select 5 candidate reference genes from the sequencing results, carry out fluorescence quantitative PCR amplification on rice cDNA template, according to the expression difference of candidate genes in different rice samples, screen to the most suitable reference gene;Use the 3 reference genes screened together to calibrate and standardize the expression of target gene, can more accurately detect the expression change of indica rice gene, ensure the sensitivity and accuracy of fluorescence quantitative PCR;Use heat stress response gene to carry out method verification, find that the relative quantitative expression value of each gene in indica rice sample is consistent with the RPKM value and TPM value of sequencing result very well.

The invention discloses a method for detecting activity of human induced pluripotent stem cell derived myocardial cell telomerase, a reference gene, a primer and probe combination and application, and belongs to the technical field of stem cell biology and molecular detection. According to the method, stably expressed 18s rRNA is taken as a reference gene, total RNA at different differentiation stages of human induced pluripotent stem cell derived myocardial cells is extracted and is subjected to reverse transcription into cDNA, then a specific primer probe combination is adopted, the gene expression quantity of telomerase reverse transcriptase and 18s rRNA is detected through Taqman fluorescent quantitative PCR, and the activity of telomerase is analyzed and evaluated in a normalized manner in combination with a 2-delta delta CT method. The method solves the problem of detection deviation caused by instability of traditional reference genes, and is simple and convenient to operate, good in repeatability and reliable in result. The method can be used for evaluating the differentiation efficiency of the human induced pluripotent stem cell derived myocardial cells, the maturity of the myocardial cells, drug screening and heart disease model research, and has important application value.

The present invention provides an internal reference gene ACT of Eremochloa ophiuroides under abiotic stress, its primers and applications, belonging to the field of molecular biology. The ACT gene in the present invention is used as an internal reference gene in the application of detecting the gene expression level of Eremochloa ophiuroides under abiotic stress, and the abiotic stress is glufosinate stress. The present invention also provides specific primers for the ACT internal reference gene. The specificity of the specific primers is strong, which can greatly improve the detection efficiency when detecting the gene expression level of Eremochloa ophiuroides and improve the credibility of the detection results.

The invention discloses a monoclonal-fluorescent protein / protease double-labeled tumor cell line and a preparation method thereof. The preparation method comprises the following steps: constructing monoclonal-fluorescent protein / protease double-labeled gene plasmids and viruses; constructing a tumor cell line; tumor cell line screening: infecting tumor cells with the constructed fluorescent protein / protease double-reference gene virus; screening out a positive tumor cell line from puromycin; planting the positive tumor cell line on a 96-well tissue culture plate to establish a tumor cell line monoclonal colony; and screening by an inverted fluorescence microscope to obtain the monoclonal-fluorescent protein / protease double-labeled tumor cell line. After 15 times of passage, the obtained cell line can still show a high-intensity fluorescence signal in in-vitro and in-vivo tumor-bearing in-vivo imaging of nude mice. The problems that tumor cells for in-vivo imaging cannot be directly observed through an in-vitro fluorescence microscope and progeny tumor cell protease gene characters are sharply lost are solved, and the labor cost and the economic cost of small animal in-vivo imaging tumor research are greatly reduced.

The invention relates to the technical field of gene diagnosis, and particularly discloses a liver cancer diagnostic kit based on a GBP2 gene and a detection method, the kit comprises an RNA extraction component used for extracting total RNA from a liver cancer or para-carcinoma tissue sample to obtain an RNA sample; the reverse transcription component is used for carrying out reverse transcription reaction on the RNA sample to obtain cDNA (complementary deoxyribonucleic acid); carrying out RT-qPCR detection on the components; the fluorescent dye premix liquid is used for carrying out real-time quantitative PCR amplification on the cDNA to obtain a GBP2 gene expression Ct (GBP2) value; the internal reference gene detection component comprises a GAPDH primer pair and is used for obtaining an internal reference Ct (GAPDH) value; the components are interpreted, delta Ct is obtained through calculation, a preset threshold value is compared with delta Ct, and the GBP2 expression level is obtained through judgment; the GBP2 is used as a liver cancer specific diagnosis marker, the detection rate of early liver cancer can be improved by detecting the expression level (such as mRNA or protein level) of the GBP2, and the gene is particularly suitable for AFP negative or low expression patients and has higher sensitivity and specificity.

The invention discloses a kit for quantitatively detecting methylation of an FOXK1 gene promoter region and application of the kit to identification of hysteromyoma and hysterosarcoma, the sequence of the FOXK1 gene promoter region is chr7: 4682295-4685295, the reference genome version is GRCh37 / hg19, and the FOXK1 gene promoter region has a nucleotide sequence shown in SEQ ID NO.1. The FOXK1 gene promoter region has a nucleotide sequence shown in SEQ ID NO.2. The FOXK1 gene promoter region has a nucleotide sequence shown in SEQ ID NO.2. The kit comprises a pair of primers FOXK1-Me-F and FOXK1-Me-R for detecting the methylation degree of the FOXK1 gene and a probe FOXK1-Me-Probe, a pair of primers FOXK1-unMe-F and FOXK1-unMe-R for detecting the non-methylation degree of the FOXK1 gene and a probe FOXK1-unMe-Probe, and a pair of primers B2M-F and B2M-R for a reference gene B2M and a probe B2M-Probe. By detecting a specific gene methylation marker of cfDNA in peripheral blood and particularly utilizing an MS-qPCR technology, the invention provides a non-invasive or minimally invasive early differential diagnosis method for uterine sarcoma, which is high in sensitivity, relatively simple and convenient to operate and controllable in cost. The method is helpful for distinguishing benign and malignant lesions before an operation, provides an important basis for clinical decision making, avoids unnecessary operation risks, and improves prognosis of a patient.

The invention discloses oligonucleotide for screening and identifying FGFR1 rearrangement by a multiplex fluorescent PCR (Polymerase Chain Reaction) technology, the oligonucleotide comprises an upstream primer, a downstream primer and a probe for detecting a ZNF198-FGFR1 fusion gene, and a kit and a detection method based on a Taqman probe real-time fluorescent quantitative PCR technology. The kit is used for rapidly and accurately detecting the expression level of the ZNF198-FGFR1 fusion gene in a patient with 8p11 myeloproliferative syndrome (EMS). A brand new oligonucleotide sequence is designed for a ZNF198-FGFR1 fusion site, and the amplification efficiency and the detection sensitivity are remarkably improved by optimizing the molar ratio of the primer to the probe; a double-standard curve quantitative technology is adopted, and a reference gene ABL and a target gene are synchronously detected, so that relative quantitative analysis of a fusion gene is realized, and variation interference among samples is avoided; due to the design of the integrated kit, sample treatment steps are simplified; the kit contains positive / negative / blank reference substances, ensures the reliability of a detection result, and is suitable for clinical trace residue monitoring and early targeted treatment guidance.