41 results about "Molecular diagnostic techniques" patented technology

This invention belongs to the field of molecular diagnostics, specifically relating to biomarkers for the diagnosis of herpes simplex keratitis (HSK) and their applications. This invention is the first to demonstrate that C1QA and FCERT1G are specific and reliable inflammation-related biomarkers for HSK. Diagnostic and assessment methods based on these biomarkers can provide crucial molecular evidence for the early detection, accurate diagnosis, objective severity grading, and personalized treatment of HSK. Simultaneously, this invention reveals the potential roles of FCERT1G and C1QA in HSK and provides new ideas for targeted therapy of this disease, particularly the application of goserelin as an inflammation modulator, offering a new direction for HSK treatment.

The application belongs to the technical field of food detection and molecular diagnosis, and particularly relates to a molecular diagnosis system for food source component identification. The system comprises a handheld shell, and an automatic sample processing unit, a molecular detection unit, a central control unit and a data transmission unit are integrated in the shell. The automatic sample processing unit comprises at least two processing modules and an automatic transfer mechanism, which are used for crushing, lysing and automatic transfer of a solid sample, and an in-situ cleaning and disinfecting assembly is integrated. The application realizes full-process automation, handholding and intelligence from a solid sample to a detection result, and provides an innovative solution for food safety on-site rapid detection and intelligent supervision.

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 application provides a microfluidic chip for diagnosing avian influenza H3 / H5 / H6 / H9 molecular typing based on microfluidics and a use method, relates to the field of pathogenic microorganism testing technology, and improves the efficiency, accuracy and convenience of avian influenza molecular diagnosis, promotes the development of molecular diagnosis technology, and uses the microfluidic chip body to be supported and used through the chip support, opens the protection layer attached to the microfluidic chip body by means of the light-proof assembly on the side, and can provide a light-proof environment again during and after the detection process, and can maintain a longer detection state.

This invention discloses a rapid detection method for MNSs blood typing based on multiplex digital PCR and a dedicated probe and primer set, belonging to the field of clinical laboratory medicine and molecular diagnostics technology. The method involves mixing a PCR reaction mix, the sample to be tested, enzyme-free purified water, M / N upstream primer, M / N downstream primer, S / s upstream primer, S / s downstream primer, upstream internal control primer, downstream internal control primer, M probe, N probe, S probe, s probe, and internal control probe to obtain a dPCR reaction system. This dPCR reaction system is then added to a microfluidic chip to generate ≥30,000 homogeneous water-in-oil droplets, followed by PCR amplification. The fluorescence signal is then detected using a droplet analyzer, achieving accurate and efficient detection of MNSs blood typing. This method addresses the shortcomings of existing molecular biological detection methods for MNSs blood typing, such as insufficient specificity, sensitivity, and throughput, as well as complex operation and high cost, providing reliable technical support for clinical transfusion safety.

This invention belongs to the field of programmable nuclease technology, and particularly relates to a TcAgo protein mutant and its encoding gene, a nucleic acid cleavage reagent containing the mutant, and its applications. The amino acid sequence of the TcAgo protein mutant is selected from one of SEQ ID NO. 2-4. The TcAgo protein mutant provided by this invention has advantages such as improved low-temperature and medium-temperature performance, high room-temperature catalytic activity, wide temperature adaptability, broadened substrate spectrum, and high stability. It is beneficial for overcoming high-temperature dependence, adapting to in vivo and room-temperature physiological environments, and promoting the development of gene editing, molecular diagnostic technology, biosensing, and other fields towards in vivo applications and mild reactions. It solves the problem of insufficient activity and limited application of thermophilic pAgo proteins, including TcAgo, under mild conditions in existing technologies.

The application belongs to the technical field of gene detection and molecular diagnosis, and particularly relates to a kit for detecting a pathogenic gene of phenylketonuria and application thereof. The kit comprises a primer pair for amplifying specific exons and splicing regions of a phenylalanine hydroxylase gene, and specific probes for detecting hot spot mutations and deletion / repetition variations of the gene. The application can detect genetic variations related to phenylketonuria in one time, quickly and accurately by combining optimized multiplex polymerase chain reaction with high-throughput sequencing or gene chip technology, and covers various known hot spot mutations and copy number variations including c.1222C>T, c.1068-11G>A, c.728G>A and c.1162G>A. The kit has high detection sensitivity and strong specificity, and is suitable for positive recall diagnosis of neonatal phenylketonuria screening, genetic diagnosis of suspected patients, carrier screening and prenatal diagnosis, and provides an efficient tool for precise prevention and control of phenylketonuria.

This invention belongs to the field of molecular diagnostic technology and discloses a premixed reagent solution suitable for multiplex fluorescent PCR systems, its preparation method, and its application. The premixed solution uses purified water as a solvent and 2 mol / L sorbitol, 25% (w / v) trehalose, 2% (w / v) glycine, 20% (w / v) dextran, and stock solution of Tween-20 as raw materials. The preparation method involves mixing the components thoroughly and then drying them in a hot air drying oven to obtain a transparent gel-like product. This product is compatible with multiplex fluorescent PCR using multiple primers and probes and multi-channel detection. It has advantages such as low hygroscopicity, good resolubility, and excellent stability. It maintains excellent amplification efficiency and detection sensitivity even after room temperature storage, requires no cold chain transportation, and is convenient to use. Furthermore, the process is simple, cost-effective, and has good batch reproducibility, making it widely applicable in nucleic acid amplification scenarios such as clinical diagnosis and food safety monitoring.

The application belongs to the technical field of molecular diagnosis, and particularly relates to extracellular vesicle lncRNA as a molecular marker for diagnosing heart failure combined with dilated cardiomyopathy. The molecular marker is NR_045681, and the specific nucleic acid sequence of the extracellular vesicle lncRNA is shown in SEQ ID NO. 5. The technical scheme provided by the application effectively improves the detection accuracy of heart failure combined with dilated cardiomyopathy. The P value of the combination of the molecular marker is less than 0.05, the |log fold change (FC)| is greater than 1, and the molecular marker has obvious differential expression relative to a healthy control group.

The application discloses a mycoplasma pneumoniae 23S rRNA gene A2063G drug resistance mutation detection method based on a CRISPR-LbCas12a system and belongs to the technical field of molecular diagnosis. The detection method can accurately distinguish single base differences such as A2063 and G2063, effectively avoids mismatch, and ensures the accuracy of results. Relying on a one-step reaction of RPA and CRISPR, the total time consumption is less than 45 minutes, and the waiting time is significantly shortened. The whole reaction is carried out under constant temperature conditions, does not need to rely on complex equipment such as a thermal cycler, is easy to operate, is very suitable for the application of primary medical institutions and instant test scenes. The cost control advantage is outstanding, the reagent cost is about 1 / 50 of second-generation sequencing and 1 / 5 of fluorescent PCR, and the economy is extremely good. In addition, the application technology shows good multiplex detection potential and wide application prospect.

This invention discloses a gene biomarker panel for detecting PCNSL based on cerebrospinal fluid (cerebrospinal fluid) ctDNA, relating to the fields of gene detection and molecular diagnostics. The gene biomarker panel includes MYD88, PIM1, CD79B, GNA13, IRF4, DTX1, KMT2D, and B2M. By detecting the mutation status of these genes in a subject's cerebrospinal fluid sample, the mutation status is input into a trained random forest classification model to generate auxiliary judgment results, providing molecular evidence for diagnosing PCNSL in the subject. This eight-gene combination panel has shown high diagnostic specificity in multiple cerebrospinal fluid ctDNA-based validation cohorts and has the ability to identify MYD88 wild-type PCNSL.

This invention relates to the fields of microfluidic chip technology and molecular diagnostic technology, and discloses a self-driven digital microfluidic chip and its application in digital recombinase polymerase amplification. The chip comprises two layers: an upper PDMS sheet and a lower glass plate. The lower surface of the upper PDMS sheet is etched with a microfluidic channel structure, including a sample loading port, a droplet reactor array, microchannels, and a pump chamber. The upper and lower layers are thermally bonded to form a sealed flow channel. Based on the gas permeability of PDMS material, this invention achieves self-driven sample introduction and digital dispensing by pre-degassing to create negative pressure, eliminating the need for any external equipment. The chip structure of this invention is rationally designed, with a simple preparation process, low cost, and convenient operation. It can automatically complete sample digitization within 5-10 minutes, supports multi-channel parallel detection, is highly compatible with RPA reaction systems, and can achieve absolute quantitative analysis of target nucleic acids. It has advantages such as high detection sensitivity, accurate quantification, and good repeatability, and has broad application prospects in the field of molecular diagnostics.

The present application relates to the field of agricultural ecological monitoring and molecular diagnosis, and specifically discloses a method for rapidly detecting soil cadmium pollution by using plant genetic markers, which aims to solve the problems of complex pretreatment, dependence on large instruments and inability to reflect biological availability in existing soil cadmium detection, selects the cadmium-specific response gene rb_592991 of the super-hyperaccumulator Bidens pilosa as a target, extracts the total RNA of the Bidens pilosa exposed to the soil to be detected, performs one-pot constant-temperature reaction at 60 DEG C by using RT-LAMP combined with CRISPR / Cas12b system, and realizes naked-eye visual interpretation by using a colloidal gold lateral flow test strip. The detection system of the present application has a minimum detection limit (LOD) of 0.2 mg / L in a water culture environment, has strong anti-interference property, does not require expensive instruments, and can rapidly evaluate the biological availability of soil cadmium in situ, thereby providing a simple, efficient and new strategy for early warning of cadmium pollution in farmland and traditional Chinese medicine production areas.

This invention provides a CRISPR-Cas12a-based nanopore sensor and its application in IGF-1 protein detection, belonging to the fields of biosensing and molecular diagnostics. The nanopore sensor includes: a solid-state nanopore unit, which is a glass capillary nanoparticle with an opening at the tip and a conductive layer covering the inner wall of the nanoparticle; a reporter molecule chemically bonded to the conductive layer; a protein recognition and signal conversion unit, including a DNA aptamer capable of specifically binding to Atlantic salmon IGF-1 protein and a single-stranded DNA complementary strand partially complementary to the DNA aptamer; and a signal amplification unit containing the Cas12a protein and crRNA complementary to the target DNA strand. This sensor achieves efficient, sensitive, and specific conversion from protein to electrical signal through a cleverly designed molecular competition system.

This invention discloses a one-step rapid nucleic acid extraction and detection multifunctional reaction assembly and its usage method, belonging to the field of molecular diagnostic technology. The assembly includes a sample loading and driving unit (syringe), a lysis filter tube, a rapid nucleic acid extraction inner tube, a lower plug, and a reaction tube pre-filled with lyophilized reagent balls. The rapid nucleic acid extraction inner tube is placed inside the reaction tube, with a permeable air channel between its outer wall and the inner wall of the reaction tube. All components are sealed together by interference fit or other methods to form a fully enclosed system. In use, the sample is driven by the syringe and flows sequentially through the lysis filter tube and the rapid nucleic acid extraction inner tube, completing lysis, impurity filtration, and adsorption; the elution buffer elutes the nucleic acid to the bottom of the reaction tube, dissolving the lyophilized reagent to form the reaction system, which can then be directly used for detection. This invention highly integrates nucleic acid extraction, purification, and amplification detection into one unit, making operation extremely simple, the entire process completely sealed to effectively prevent contamination, and the cost low, making it particularly suitable for point-of-care testing (POCT) scenarios.

The present application belongs to the technical field of molecular diagnosis, and relates to a gene combination for evaluating prognosis of multiple myeloma and PRMM risk and a method for evaluating the same. The gene combination is a combination of MCM3, FABP5, MIR4435-2HG, FH, HELLS, CCDC34, RPS6KA1, FEN1 and OPN3 genes. Detection with the gene combination as a biomarker can specifically and accurately evaluate the prognosis of a multiple myeloma patient, especially the death risk, and the risk of the multiple myeloma patient being a primary refractory multiple myeloma according to the detection result.

This invention relates to a triple nano-PCR detection method for silkworm microsporidia, Beauveria bassiana, and nucleopolyhedrovirus, belonging to the field of molecular diagnostic technology. Addressing the technical problem of existing silkworm disease diagnostic methods' inability to simultaneously, efficiently, and sensitively identify these three commonly mixed infection pathogens, this invention provides an integrated solution. The key technical points include extracting DNA from the sample to be tested, using this DNA as a template, performing nano-PCR amplification in the same reaction system using a specific primer set, and subsequently detecting the amplification products by agarose gel electrophoresis. The infection status of silkworm microsporidia, Beauveria bassiana, and nucleopolyhedrovirus is determined based on the specific bands at 733 bp, 230 bp, and 860 bp, respectively. This method is mainly used for the rapid, simultaneous diagnosis and identification of mixed infections of microsporidia, Beauveria bassiana, and hemorrhagic septicemia during silkworm rearing.

This invention discloses a probe for detecting microRNA-21 based on a bioorthogonal cyclic amplification reaction, its synthesis method, and its applications, belonging to the field of biomedical detection and molecular diagnostics technology. The probe IDCR comprises compounds RNA-Cy3Tz and RNA-ABN; this probe can achieve signal amplification through catalytic target cycling, with each miRNA molecule achieving a turnover rate of 2.2 × 10⁻⁶. 9 The detection limit is as low as 3.58×10 ‑18 This probe exhibits high specificity and a wide linear range of 100 aM to 100 nM. It can perform in-situ, real-time imaging in living cells and tissues without lysing cells, and can also be used for clinical serum sample detection, effectively distinguishing miRNA levels between lung cancer patients and healthy individuals. It holds significant promise for applications in non-invasive diagnostics and biomedical research.

This invention relates to the field of biomedical detection and molecular diagnostics, specifically to the application of KYAT1 protein and its modification sites as biomarkers in the preparation of products for the auxiliary diagnosis and prognostic assessment of breast cancer. The invention aims to reveal the positive correlation between high expression of the KYAT1 protein and the hydroxylation status of its proline at position 352 with the malignant biological behavior of breast cancer, and to use this as a molecular biomarker for the preparation of products to aid in the diagnosis of breast cancer or assess its perioperative prognostic risk.

This invention discloses an exosome-based tumor diagnostic kit and its applications, relating to the fields of medical laboratory science and molecular diagnostics. The kit includes: exosome isolation components; exosome lysis and nucleic acid extraction components; internal standard and quantitative control components; reverse transcription components; molecular detection components; protein detection components; and interpretation and analysis components. This invention achieves complementary fusion of nucleic acid and protein information by simultaneously detecting exosome nucleic acid markers and exosome-derived protein markers from the same blood sample and employing a pre-defined normalization and weighted interpretation algorithm. This fusion strategy overcomes the problem of decreased sensitivity or specificity caused by biological heterogeneity or sample batch variations of single markers, making the identification of early or minor lesions more robust. Simultaneously, through synergistic interpretation among multiple markers, false positive signals originating from benign diseases or physiological fluctuations can be effectively suppressed, thereby improving the reliability of clinical screening and diagnosis.

The application discloses a marker tiRNA-Gln-TTG-001 and application thereof, and belongs to the technical field of biotechnology and molecular diagnosis. The applicant finds that, compared with lung cancer patients without brain metastasis, tiRNA-Gln-TTG-001 is significantly highly expressed in the peripheral blood circulation of lung cancer patients with brain metastasis, which provides an important basis for the marker as a specific plasma marker for predicting, monitoring or diagnosing lung cancer brain metastasis. The new clinical cases are selected for verification, and the results show that tiRNA-Gln-TTG-001 can be used as a specific metastasis marker for lung cancer brain metastasis, which opens up a new non-invasive, convenient and specific detection way for early prediction, dynamic monitoring, diagnosis and efficacy evaluation of lung cancer brain metastasis.

This invention discloses a gene composition, kit, and application for early diagnosis of renal cell carcinoma, relating to the field of molecular diagnostic technology. The gene composition consists of the PLIN2 gene, SLC25A25 gene, PITX1 gene, and SOX1 gene. The differential methylation site of the PLIN2 gene is cg13990947, the differential methylation site of the SLC25A25 gene is cg14294859, the differential methylation site of the PITX1 gene is cg02037307, and the differential methylation site of the SOX1 gene is cg1. 6705627; This invention, by screening four genes (PLIN2, SLC25A25, PITX1, and SOX1) and specific methylation site combinations, can accurately identify two major subtypes of renal cell carcinoma—clear cell and papillary renal cell carcinoma—compared to single-gene detection. The methylation difference between cancerous and normal tissues, Δβ-value, is greater than 0.2, solving the problem of heterogeneous renal cell carcinoma detection. The optimized urine free DNA lysis buffer and bisulfite conversion system improve the nucleic acid extraction and conversion rates. Combined with dedicated methylation-specific primers and probes, it enables non-invasive detection of urine samples.

This invention relates to the field of biological sample pretreatment and molecular diagnostics, and discloses a cell-free DNA extraction device and method based on electrophoresis-dialysis coupling. The device includes an outer container, a middle container, an inner container, and a collection container. The middle container is disposed within the outer container, and a gap exists between the middle and outer containers to form a first electrode cavity, within which an anode is located. The inner container is disposed within the middle container, and a gap exists between the inner and middle containers to form a second electrode cavity, within which a cathode is located. The collection container has open ends, with a connection end and a filter end from top to bottom. The connection end is detachably connected to the middle container and communicates with the inner container. The filter end communicates with the first electrode cavity and is equipped with a dialysis membrane. This invention enables cell-free DNA to achieve directional migration, desalting, and enrichment under an electric field, and is suitable for rapid cfDNA extraction from complex biological samples.

This invention belongs to the field of biotechnology and molecular diagnostics. Addressing the limitations of existing technologies in gastric cancer diagnosis, such as complex procedures, low patient compliance, and high costs, this invention proposes a circRNA biomarker for gastric cancer diagnosis and its application. The circRNA biomarker of this invention is hsa_circ_0001185. Quantitative analysis of hsa_circ_0001185 confirms its significant high expression in the serum of gastric cancer patients and its close correlation with clinical parameters such as tumor differentiation degree, lymph node metastasis, and TNM stage. It can significantly distinguish between gastric cancer patients, gastritis patients, and healthy individuals, exhibiting high sensitivity and specificity, thus providing a new solution for gastric cancer screening and auxiliary diagnosis.

The application discloses a method for improving the sensitivity of RPA-CRISPR-Cas12a detection and application thereof in one-tube detection of chikungunya virus, and relates to the technical field of biology.The full-length crRNA is reformed to form a truncated crRNA (Truncate crRNA), the non-key region is accurately shortened, the space-time regulation of Cas protein activity is realized, and the truncated crRNA plays a key role in CRISPR molecular detection (such as an RPA-CRISPR-Cas12a system). The method can significantly improve the specificity of RPA-CRISPR-Cas12a detection, effectively avoid cross-reaction, inhibit non-specific early activation, significantly reduce background signals, and provide an optimization strategy for developing a molecular diagnostic technology with higher sensitivity.

This invention belongs to the field of molecular diagnostics technology, and particularly relates to a reaction control system for automated NGS library construction. The system includes a base plate, a motion module, a top plate, a nucleic acid extraction module, a nucleic acid fragmentation module, a first driving component, a library amplification module, and a processor. The motion module is mounted on the base plate, and the top plate is directly or indirectly connected to the motion end of the motion module. Multiple first driving components are disposed on the top plate. The motion module and the first driving components can move the nucleic acid extraction module and the nucleic acid fragmentation module relative to the base plate in different directions to drive them to engage with corresponding reaction wells. The library amplification module is disposed on the base plate and provides the required temperature for the amplification reaction to the corresponding reaction wells. This invention, in conjunction with a reaction apparatus, can automate NGS library construction. The modular design facilitates adjustment, and the library amplification module's heat dissipation accelerates cooling and shortens reaction time.

The application discloses a method for determining human bocavirus type I / II based on HNB-LAMP and CRISPR / Cas12a-AuNP double colorimetric signals, and belongs to the technical field of in vitro molecular diagnosis. The application first performs isothermal amplification on a human bocavirus NP1 gene in a single tube by using HNB-LAMP technology, realizes one-step visual preliminary screening through color change (purple to blue) of hydroxynaphthol blue (HNB), then adds a CRISPR / Cas12a-AuNP typing system to positive amplification products, utilizes trans-cleavage of HT44 hairpin transduction structure after Cas12a is activated by specific amplicons, releases RNA cross-linking agent, and then causes aggregation and precipitation of DNA functionalized gold nanoparticles (AuNPs), to produce visible purple precipitate. The application realizes accurate typing detection of human bocavirus type I / II through double colorimetric signals (blue solution + purple precipitate), does not need complex instruments, is simple and easy to operate, has high sensitivity and strong specificity, and is suitable for primary medical units and on-site rapid screening.

The present application relates to the technical field of molecular diagnosis, and specifically relates to application of female uterine cavity flora as a marker in endometriosis diagnosis. The present application collects uterine cavity flora of patients, extracts genomic DNA, performs bacterial 16s-rRNA sequencing, uses KEGG as a reference database for function prediction, determines that abnormal abundance of uterine cavity flora dominant bacteria has significant correlation with dysmenorrhea and high CA125 value of endometriosis patients, clarifies the correlation between abnormal uterine cavity flora and endometriosis, and provides a new method for evaluating the risk of endometriosis by detecting uterine cavity flora atlas.