37 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 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 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 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 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 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 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 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 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 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 application particularly relates to a primer probe, a kit and application for detecting the GJB2 gene copy number of a non-syndromic hearing loss patient by using microdroplet digital PCR. The primer probe combination provided by the application comprises a target gene detection primer pair, a target gene detection probe, an upstream primer of a reference gene, a downstream primer of the reference gene and a reference gene probe, and the sequence information is shown as SEQ ID NO. 6-11; and the application further provides a kit for detecting the GJB2 gene copy number of a non-syndromic hearing loss patient. The primer probe combination and the kit provided by the application are applied to the detection of the GJB2 gene copy number variation of a non-syndromic hearing loss patient, and the accuracy of the detection of genetic hearing loss is significantly improved; according to the determination result, it is determined whether the GJB2 gene expression is abnormal or not, the disease prognosis is evaluated, potential therapeutic drugs are screened, genetic counseling services are provided or individualized medical schemes are formulated, and the application has a wide application prospect.

The present application relates to the field of C12Q1 / 6851, and more particularly, the present application relates to a primer probe combination, a kit and a method for detecting human ALK and ROS1 gene fusion. The primer probe combination comprises at least a primer probe combination A which specifically detects at least one EML4-ALK fusion gene; a primer probe combination B which specifically detects at least one ROS1 fusion gene; and a primer probe combination C which specifically detects an internal reference gene. The primer probe combination and the detection method of the present application have high specificity and sensitivity, improve the signal-to-noise ratio, have good resolution and accuracy, and have excellent linear relationship, so that the result can be easily judged.

The invention discloses application of an Actin gene and / or an RPL35 gene in preparation of a reference gene for quantitative analysis of gene expression in a camellia oleifera seed kernel development stage. Candidate genes which are stably expressed in different development stages are screened based on transcriptome data, the expression stability of the candidate genes is evaluated through qRT-PCR in combination with a plurality of stability evaluation methods (geNorm, NormFinder, BestKeeper, Delta-Ct and RefFinder) systems, and finally Actin and RPL35 are determined as optimal reference genes. The invention also provides a specific primer of the reference gene, and the specific primer has the characteristics of high amplification efficiency, strong specificity and the like, is suitable for standardized detection of the expression quantity of the target gene in the camellia oleifera seed kernel development stage, and can effectively improve the accuracy and repeatability of gene expression analysis; and a stable and reliable molecular tool is provided for subsequent grease accumulation and functional gene research.

The invention provides an automatic single cell annotation method and system, and relates to the technical field of automatic cell type annotation. According to the method, a reference gene set is constructed based on marker genes of a reference map, a weight rule is set, and a probability score and confidence of a cell type are calculated in combination with a semi-supervised, unsupervised or predefined module mode. Through multi-algorithm integration and reannotation rechecking, annotation precision and consistency are optimized, final labels and related evidences are output, and data tracing and reanalysis are supported.

The invention relates to the technical field of molecular biology and environment, and discloses a primer group, a kit and application thereof, and a method for detecting biotoxicity of an environmental sample, the primer group comprises a primer pair for specific amplification of an hsp70.3 gene and an mt2 gene, and a primer pair for specific amplification of a reference gene; the primer pair for specifically amplifying the hsp70.3 gene comprises an upstream primer with a nucleotide sequence as shown in SEQ ID NO.3 and a downstream primer with a nucleotide sequence as shown in SEQ ID NO.4; the primer pair for specifically amplifying the mt2 gene comprises an upstream primer with a nucleotide sequence as shown in SEQ ID NO.5 and a downstream primer with a nucleotide sequence as shown in SEQ ID NO.6. The primer group disclosed by the invention can be used for monitoring the content change of hsp70.3 and mt2 in a living body, so that the biotoxicity of a to-be-detected environmental sample can be analyzed, the early warning of heavy metal pollutants is realized, particularly the existence of the heavy metal pollutants in the to-be-detected environmental sample can be prompted, and the heavy metal detection can be further combined for analysis in the later period.

The application relates to the fields of molecular biology and environmental biotechnology, and discloses a PCR chip, a kit and application thereof. The PCR chip comprises a chip plate, primer pairs for specifically amplifying bvcA genes, cfrA genes, mbrA genes, pceA genes, pteA genes, tceA genes, vcrA genes and internal reference genes are loaded on the chip plate, and a gradient dilution template of an internal reference gene for drawing a standard curve is arranged on the chip plate; the gradient dilution template of the internal reference gene is obtained by gradient dilution of internal reference gene template DNA and addition of internal reference gene primers. The PCR chip can detect absolute quantification of chlorinated hydrocarbon microbial degradation related reduction dehalogen functional genes through qPCR technology, so that the PCR chip can be used for accurately detecting the copy number of the chlorinated hydrocarbon microbial degradation related reduction dehalogen functional genes, thereby the feasibility of chlorinated hydrocarbon pollution bioremediation can be explored, the chlorinated hydrocarbon pollution bioremediation process can be evaluated, and the chlorinated hydrocarbon pollution bioremediation method can be further assisted and guided.

The invention discloses a methylation marker combination for early screening of endometrial cancer, and belongs to the cross technical field of molecular diagnosis and bioinformatics. Three methylation markers CDO1, NKX2-6 and SORCS3 are screened out through bioinformatics analysis, NGS targeted sequencing and qPCR verification, specific primers SEQ ID NO.1-2, 4-5 and 7-8 and probes SEQ ID NO.3, 6 and 9 of the methylation markers are designed, and a fluorescent quantitative PCR detection system is constructed in combination with a reference gene BACT. According to the method, the methylation level of a three-gene promoter region in a sample is detected, the cutoff value is 0.36, the sensitivity of endometrial cancer diagnosis of cast-off cells is 90.37%, the specificity is 89.60%, AUC is equal to 0.9682, and benign and cancerous samples can be effectively distinguished. According to the invention, noninvasive / minimally invasive early screening of endometrial cancer is realized, diagnosis accuracy and safety are improved, and NKX2-6 and SORCS3 are newly discovered markers and have important scientific value and clinical transformation potential.

The invention discloses a method for detecting a bulbus fritillariae cirrhosae medicinal material based on Taqman fluorescent probe double-marking linear correction quantitative PCR (Polymerase Chain Reaction), which comprises the following steps: amplifying a test sample and a standard reference substance in a PCR reaction system based on bulbus fritillariae cirrhosae specific primers and probes designed on the basis of six bulbus fritillariae cirrhosae-based original plant conserved gene sequences, and 18sRNA universal reference gene primers and probes; the method comprises the following steps: qualitatively determining whether a test sample contains the bulbus fritillariae cirrhosae component or not according to amplification results of a bulbus fritillariae cirrhosae specific primer and a probe, taking a standard reference substance as a template to obtain two standard curves of a bulbus fritillariae cirrhosae specific gene and a 18sRNA gene, and finally quantitatively analyzing the percentage content of the bulbus fritillariae cirrhosae component of the test sample by adopting a double-marking linear correction method. According to the method, the bulbus fritillariae cirrhosae components can be accurately and reliably quantified, bulbus fritillariae cirrhosae and non-bulbus fritillariae cirrhosae medicinal materials are effectively identified, the detection universality of all certified bulbus fritillariae cirrhosae bases specified by pharmacopeia is ensured, and missing detection or false detection is avoided.

The invention discloses an NPM1 mutation dynamic monitoring method based on a microdroplet digital PCR technology, and relates to the technical field of molecular diagnos.The method comprises the steps that S1, RNA is extracted from a sample; s2, carrying out reverse transcription on the extracted RNA to obtain cDNA (complementary deoxyribonucleic acid); s3, designing a specific primer and a fluorescent probe aiming at the NPM1 gene exon 12 A type mutation, and designing a primer and a fluorescent probe of an ABL1 gene at the same time; s4, verifying the designed primer and probe and confirming the performance of a reaction system; s5, preparing a reaction mixed solution; s6, carrying out oil drop generation on the reaction mixed solution; specific primers and a fluorescent probe are designed for NPM1 gene exon 12 type A mutation, ABL1 is used as a reference gene, a droplet digital PCR platform is used for detection, the amplification efficiency and specificity of the primers and the probe are verified through real-time fluorescent quantitative PCR, the concentration combination of the primers and the probe is optimized by using a clinical sample, and the kit is used for detecting the NPM1 gene exon 12 type A mutation. And a plurality of samples are adopted for cross validation, so that high sensitivity and specificity of the method on NPM1 mutation detection are ensured.

The invention belongs to the technical field of microbial molecule detection, and particularly relates to a multiple LAMP mycobacterium tuberculosis complex group detection method based on a probe method, the LAMP multiple probe method is used for detecting mycobacterium tuberculosis complex flora IS6110 and IS1081 primer groups, the mycobacterium tuberculosis complex flora can be conveniently and rapidly detected at the same time, the positive detection rate of a sputum sample is increased, and the detection time is shortened. Meanwhile, non-specific amplification is weakened, false positive is reduced, the arrangement of the reference gene GAPDH can effectively monitor the sample collection / sample preservation / sample nucleic acid extraction process, and false negative is avoided; the specificity is high, and cross reaction with common strains is avoided. And the sensitivity can reach 10 < 0 > cop i es / u < l >.