115 results about "Sputum sample" patented technology

The invention discloses a medical self-sputum smear dyeing device and method instead of artificial operation in the tuberculosis detection technique domain, which comprises the following parts: three-dimensional right-angle coordinate mechanical console, cam, smear operation head, dyeing liquor sprayer, sampling tag box component, sample box location board, slide location rack, pusher box, creeping pump group, and display and operation button. The method comprises the following steps: finishing automatic box opening, shooting and image display; selecting the suspected part of sputum sample artificially; gathering the sample to coat on the slide; dying and drying fractionally; preparing the smear sputum sample to satisfy the microscopic requirement. The invention prevents the detecting personnel from infection and medical pollution, which improves the quality of sputum smear and shortens the sample disposal time.

The invention relates to a method for storing integrity of nucleic acids (DNA and RNA) in a sputum sample in the field of biology. The invention provides phlegm paper, which can be used for conveniently storing nucleic acids in the sputum sample at room temperature; nucleic acid protecting liquid is infiltrated on the phlegm paper, and can be used for inactivating nuclease in the sputum sample; DNA and RNA in the sputum are effectively prevented from being degraded, so as to store and transport the sputum sample. At room temperature (25-35 DEG C), the nucleic acids in the sputum sample can be stably kept for 7-14 days. Meanwhile, the invention also provides a corresponding nucleic acid extraction method.

The invention belongs to the field of biomedical detection, relates to the technical field of microbe molecule biology detection, in particular to a multiplex polymerase chain reaction (PCR) kit capable of quickly identifying and diagnosing mycobacterium tuberculosis, nontuberculous mycobacteria, mycobacterium bovis and Bacillus Calmette Guerin vaccine simultaneously, and discloses four primer pair sequences designed for specific segments of four differential genes, namely 16S rRNA, IS6110, Rv2652 and Rv3873 of the mycobacterium, and amplification conditions for multiplex PCR. By the invention, a mycobacteria tuberculosis complex sequence can be quickly and accurately amplified in a sputum sample of a tuberculosis patient, and species identification can be carried out; the sensitivity is 50 colony forming units/template, which is far higher than that of an acid-fast staining method, an inspection procedure for the mycobacterium tuberculosis is greatly simplified, and a detection speed is improved; and meanwhile, different mycobacterium strains can be identified through once PCR, and a technical means is provided for clinic quick diagnosis of tuberculosis and molecular epidemiology survey.

A drug-resistance gene film chip for detecting mycobacterium tuberculosis is prepared by designing 54 probe spotting on a nylon film by aiming at the mutant sites of mycobacterium tuberculosis rpoB, kagG, embB, inhA, ahpC, gyrA, rrs, rpsL and pncA gene; 12 pairs of specific primers with biotin-labeled at 5' end are utilized; a sample DNA is subjected to triple PCR and amplified to form a large amount of gene fragment products with biotin; the amplified products and the probes on the film chip carry out specific hybridization; and then film washing, enzyme-linking and color reaction are carried out, thus preparing the chip. The gene film chip and the detection method thereof can detect the common gene mutations of the mycobacterium tuberculosis on the drug resistance of drugs such as isoniazid, rifampicin, streptomycin, ethambutol, pyrazinamide, quinolone and the like at one step, and are applicable to extracorporeal detection sputum sample, clinical isolation strains and mycobacterium tuberculosis multi-drug resistant gene in the organization sample.

The invention provides a method for quickly detecting klebsiella pneumoniae triple qPCR in a respiratory tract sample. The method comprises the following steps: collecting and selecting a clinical respiratory tract sample; performing bacterial culture and identifying; extracting and purifying nucleic acid; constructing quality control/interior label plasmid; designing and compounding a sequencing primer; optimizing a clinical isolate high-fidelity system, amplifying and judging a result; sequencing a high-fidelity PCR product and controlling quality; designing a triple qPCR primer and a probe and compounding a marker; and optimizing a triple qPCR system, amplifying and judging the result. The method has the beneficial effects that klebsiella pneumoniae rcsA and 23s RNA housekeeping genes can be simultaneously detected, and the adopted primer and probe are designed in the manner of effectively avoiding a variation area or a mutational site according to a gene sequencing result of popular distribution strain in China; a quick sputum liquefying bactericide is added into a sputum sample, so that the sputum can be quickly liquefied, pathogenic microorganisms can be killed, and an operator can be protected from being infected; and compared with the traditional method, the method provided by the invention can reduce the time by about 1 hour and has the characteristics of simple and convenient operation, high speed, safety and sensitivity.

The invention relates to an experimental device in the field of medicine and biology and particularly relates to an automatic oscillating and uniform mixing device. The device comprises a base, a rocking plate and a rocking box. The rocking plate is arranged on the base. The rocking box is arranged on the upper end surface of the rocking plate and is detachably connected to the rocking plate. Thebase comprises a motor, a driving wheel, a driven wheel and a transmission shaft. The transmission shaft is connected to an oscillating structure. The oscillating structure comprises an eccentric shaft and an eccentric wheel. The lower end surface of the rocking plate is provided with a circular groove and a first annular groove, the wall of the circular groove is provided with a through hole, a raised part and a pushing plate are arranged in the through hole, a second annular groove is arranged around the circular groove, an air bag is arranged in the first annular groove, the eccentric shaftis rotatingly connected to a connection rod, a free end of the connection rod is provided with a cylindrical raised block, the cylindrical raised block is slidingly connected to the interior of the second annular groove, the lower end surface of the base is provided with a support block, a vertical through hole is arranged in the support block, and a gas-guide tube is connected between the vertical through hole and a gas inlet of the air bag. The experimental device can automatically oscillate and mix the sputum samples in batches.

The invention relates to the field of clinical microbiological assay and provides a quick bacterium identification kit for a sputum sample and a detection method for the same. The kit comprises two universal primers for propagating the V1 variable regions of the 16S rRNA of a bacterium, two specific primers for shielding the variable regions of the 16S rRNA of a neisseria bacterium, two specific primers for shielding the variable regions of the 16S rRNA of a veillonella parvula, a sequencing primer for sequencing the positive segment of the V1, and an avidin-marked magnetic ball, wherein the sequencing result is compared with a database to determine the species of the bacteria. By applying the kit, the bacteria in a sputum sample can be distinguished by once sequencing, thus, the kit can be applied in the clinical diagnosis on the bacterial infection in the sputum sample. The invention wins precious rescuing time for a clinical treatment and has the advantages of low cost, convenient operation and high specificity.

One embodiment of the present invention provides for a method of determining if a sputum sample contains dysplastic or carcinomic cells by obtaining a sputum sample containing cells. The sputum sample is labeled with TCPP to stain cells suspected to be dysplastic or carcinomic. The labeled sputum sample is excited with an excitation wavelength of light of about 475 nm+/−30 nm and emission at about 560 nm+/−30 nm is detected from cells identified to be macrophages. An imager focuses on the plasma membrane of one or more cells suspected to be dysplastic or carcinomic and emission at about 655 nm+/−30 nm, if present, is detected for TCPP labeled cells of the sputum sample after focusing on the plasma membrane of the cells of the sputum sample. Photon flux for each pixel of a sensor is measured to obtain a value for the imaged cell. The measured value is scored to determine if a cell is cancerous or dysplastic.

The invention provides a kit for detecting SARS-CoV-2 novel coronavirus nucleic acid at a constant temperature by using an enzyme digestion probe. The kit comprises an isothermal amplification primerfor detecting an ORF1ab gene of SARS-CoV-2 novel coronavirus and a corresponding RNA-containing base probe capable of being digested by RNase H, wherein the left end and the right end of an RNA base of the base probe are respectively marked with a reporter fluorophore and a quencher. By utilizing the efficient isothermal amplification primer and the probe, constant-temperature multiple detection can be carried out on the SARS-CoV-2 novel coronavirus, and the kit has high sensitivity and high specificity. The novel coronavirus nucleic acid detection kit disclosed by the invention has high sensitivity and specificity, can be used for detecting throat swabs, alveolar lavage fluid and sputum samples of suspected novel coronavirus pneumonia cases, suspected aggregated case patients and other patients needing novel coronavirus infection diagnosis or differential diagnosis, and has the characteristics of simplicity, convenience, quickness and accuracy.

The invention discloses a real-time fluorescent quantitative PCR method used for detecting Mycobacterium tuberculosis. The method comprises the following steps: 1, collecting and processing a patient sputum sample; 2, extracting the DNA of the sample; 3, carrying out a real-time fluorescent quantitative PCR reaction of the DNA of the sample through treating the DNA of a standard strain as a positive control and disinfected water as a negative control; and 4, judging the detection result of the sample according to a Ct value obtained through the reaction, and calculating to obtain the copy number of the Mycobacterium tuberculosis in the sample. The invention also discloses a primer and a probe used in the detection method, and a kit containing the primer and the probe. The detection method has the advantages of simplicity, rapidness and high specificity, and can realize the rapid and quantitative detection of the Mycobacterium tuberculosis in clinic sputum samples, so the detection method is helpful for the prevention and the timely prevention of diseases.

The present disclosure diagnoses mycobacterium tuberculosis, not mycobacterium tuberculosis complex only. Specific target genes are selected from regions of difference of mycobacterium tuberculosis. After hybridization with labeling substances, sputum samples are processed through color developments separately for obtaining images to be analyzed automatically. Thus, the present disclosure rapidly diagnoses mycobacterium tuberculosis in the sputum samples with a simple operation and a low cost.

The present invention provides a kit for rapid detection of Mycobacterium tuberculosis in a sputum sample, and the kit includes a liquefied solution, a lysis solution, a probe solution, a stop solution, a fixative solution, a carrying glass sheet and a cover glass sheet. After liquefaction of the sputum sample, cell lysis, probe hybridization and fluorescence fixing and other operations are performed, and a fluorescence microscopy is used for macroscopic observation, counting and analysis of the Mycobacterium tuberculosis. The kit is simple in operation method, short in procedure, easy to operate, strong in test result specificity, high in sensitivity, clear in results and high in reliability, and can be used for direct, simple and effective detection of the Mycobacterium tuberculosis in the sputum sample.

The invention relates to the use of a cell-free nucleosome as a biomarker in a sputum sample for the diagnosis or detection of cancer, adenoma, autoimmune disease or inflammatory disease. The invention also relates to methods of detecting said cell free nucleosomes in sputum samples, in particular in methods of diagnosis.

The invention discloses an automatic sputum liquefying instrument and a sputum liquefying method thereof. According to the automatic sputum liquefying instrument and the sputum liquefying method thereof, in the using process, a moving arm and a mechanical arm are arranged, the moving arm is used for moving a pipette, the mechanical arm moves up and down to pump liquid, discharge the liquid and blowing and mixing the liquid uniformly, automatic sputum liquefying, sputum-culture automatic sample adding and automatic sputum liquidation liquid sub-packaging are achieved, the automatic sputum liquefying instrument can treat sixteen sputum samples within 30 min, working efficiency is improved greatly, and the instrument has the advantages that automatic operation and flux treatment are achieved, operation is easy, use is convenient, and cross contamination between the samples is avoided; direct contact can be avoided, so that the probability of personnel infection and cross-contamination of the samples is reduced, and sputum is treated fast, economically and easily. The urgent basic requirement of a clinical testing mechanism is met. The instrument is applicable to the field of medical instruments.

The invention provides a sample treatment reagent and a kit for detecting mycobacterium tuberculosis nucleic acid, and a nucleic acid amplification method of mycobacterium tuberculosis. The sample treatment reagent comprises digestive juice, a Tris-HCl solution and a lysate, wherein the digestive juice is an aqueous solution containing the following concentration components: 13-14 g/L of dithiothreitol, 100-105 g/L of sodium chloride, 2-3 g/L of potassium chloride, 14-15.5 g/L of disodium hydrogen phosphate and 2-3 g/L of potassium dihydrogen phosphate; the lysate is an aqueous solution containing the following concentration components: 0.08-0.15 w/v% of NaOH and 0.02-0.03 w/v% of SDS. The sample treatment reagent for detecting mycobacterium tuberculosis nucleic acid can extract mycobacterium tuberculosis DNA from a sputum sample more quickly and effectively, interference of human-derived DNA is effectively removed, mycobacterium tuberculosis is completely inactivated in the treatmentprocess, the risk of personnel infection is effectively reduced, and the sample treatment reagent is safer and more efficient. The kit for detecting the mycobacterium tuberculosis nucleic acid has good sensitivity, specificity and accuracy, and can realize rapid detection of the mycobacterium tuberculosis nucleic acid.

The subject invention is directed to the measurement of elastic fiber breakdown products in human sputum obtained from patients with emphysema and other lung diseases. Measurement of elastic fiber breakdown products may be used to monitor the progression of lung disease or to assess the efficacy of a treatment for lung disease. Such breakdown products are associated with lung injury resulting from degradation of lung elastic fibers. This process may occur in emphysema and other inflammatory diseases of the lung where excess amounts of elastase are secreted by inflammatory cells. The methods for measuring elastin breakdown products in certain tissue fluids (i.e. blood, urine, bronchoalveolar lavage fluid) are well known to the art but have not been previously applied to sputum samples. The use of sputum has several advantages over other fluids, including greater specificity and ease of procurement.

The invention discloses an immunoassay reagent kit based on a flexible core-shell quantum dot coupling marker and an application method of the immunoassay reagent kit, belongs to the technical field of immunoassay, and solves the technical problems that a detection kit based on the quantum dot marker and an antibody molecules are biologically marked through the ways such as electrostatic adsorption effect and direct covalent linkage, the sensitivity of the immunoassay is low and the specificity is poor in the prior art. The reagent kit comprises protein, an EDC condensating agent, a core-shell quantum dot, a first antibody of the to-be-detected antigen, and a microporous plate covering a second antibody of the to-be detected antigen, wherein the first antibody of the to-be-detected antigen can be specifically combined with the protein. The reagent kit is high in sensitivity and specificity, simple and sensitive to use, suitable for detecting samples such as blood samples, urine samples and sputum samples and applicable to the fields such as food security, environment monitoring, medical diagnosis, and sanitation epidemic prevention; the detection result is relatively clear.

The present invention relates to a novel bioactivity testing structure for single cell tracking using a gelling agent and a bioactivity testing system including the testing structure. The present invention also relates to bioactivity testing, drug susceptibility testing, antibiotic screening, and diagnostic methods using the testing structure. The bioactivity testing structure of the present invention enables very rapid and simple drug susceptibility testing of bacteria, particularly Mycobacterium tuberculosis, drug screening, and bacterial diagnosis. Particularly, the use of the testing structure enables DST and diagnosis of bacteria only by pretreatment without the need to concentrate human sputum samples irrespective of inoculum effect, ensuring rapid, accurate, and simple testing compared to conventional tuberculosis diagnosis or DST systems. In addition, the testing structure of the present invention simultaneously enables the diagnosis and drug susceptibility testing of tuberculosis. Therefore, the present invention provides an effective alternative to the prior art.

The invention provides a sputum preserving fluid. The sputum preserving fluid comprises guanidine salt, a metal chelating agent, a surfactant and the like. The preserving fluid does not contain a strong base component, so that damage to nucleic acid, particularly RNA, is reduced. The preserving fluid can keep a sputum sample stable, rapidly liquefy sputum and inhibit RNA enzyme; and RNA with relatively high purity is obtained, and the detection accuracy is improved. The liquefied sputum sample is directly subjected to RNA extraction, and the yield and purity are superior to those of a comparison method. The sample preserved by the preserving fluid can be used for various methods such as a paramagnetic particle method, a column extraction method, a cracking method and the like, and interference is avoided. The liquefied sputum sample can be directly used for cytological detection, and influence on cell density or cellular staining is avoided.

The invention provides a probe for detection and particularly relates to a probe, reagent kit and method for detecting bacteria in a sputum sample. The reagent kit comprises a liquefaction solution, a lysis solution, a probe solution, a stop solution, an immobilization solution, slide glass and cover glass. According to the probe, reagent kit and method for detecting bacteria in the sputum sample, after the sputum sample is liquefied, cell lysis, probe hybridization, fluorescence immobilization and the like are performed, and bacteria are observed by naked eyes, counted, analyzed and the like through a fluorescence microscope. The reagent kit is simple in operation method, brief in program and easy and convenient to operate, the detection result is strong in specificity, high in sensitivity, clear and high in credibility, and the probe, the reagent kit and the method can be used for directly, easily, conveniently and effectively detecting bacteria in the sputum sample.

The present invention relates to the use of nucleic acid methylation and methylation profiles to detect risk of developing neoplasia and in particular, lung cancer. The invention relates to methods for identifying a methylation profile of the CDO1, SOOX17, HOXA7, HOXA9, TAC1, and ZFP42 genes from plasma and sputum samples.

The invention discloses a novel preserving fluid for microbial nucleic acid in sputum. The preserving fluid comprises the following components in percentage by mass: 15-30% of nuclease inhibitor, 1.5-5% of a pH value stabilizer; 0.8%-1.5% of a chelating agent; 0.05%-0.15% of a mucus dissolving agent; 0.1 to 0.5 percent of sodium dodecyl sarcosinate; 10 to 20 percent of isopropanol; 2 to 15 percentof glycerol; the balance of sterilized water. The novel preserving fluid for microbial nucleic acid in sputum can rapidly dissolve and stabilize sputum samples, effectively inhibit nuclease activityand prevent cell lysis, has good sample antibacterial property, long preservation time, relatively simple components, capability of keeping nucleic acid at room temperature, and suitableness for wideclinical application.