32 results about "M. pneumoniae" patented technology

The invention provides a kit for detecting drug-resistant mutation of mycoplasma pneumoniae based on RPA-CRISPR and application of the kit. The invention provides an RPA primer pair and a crRNA sequence for detecting the drug-resistant mutation A2063G site of the mycoplasma pneumonia 23s rRNA gene, an RPA-CRISPR / Cas13a detection system is established, the detection system formed by the primer pair and the crRNA can accurately detect the drug-resistant mutation A2063G site of the mycoplasma pneumonia 23s rRNA gene within 30-40 minutes under an isothermal condition, the sensitivity and the specificity are very high, and the detection system can be used for detecting the drug-resistant mutation A2063G site of the mycoplasma pneumonia 23s rRNA gene. Amplification detection can be completed only by performing isothermal amplification at the temperature of 39 DEG C without using a variable-temperature instrument like PCR (Polymerase Chain Reaction); in addition, a lateral flow immunochromatography test strip is further combined, so that the on-site instant detection of the mycoplasma pneumoniae drug-resistant mutation can be realized.

This application discloses a Mycoplasma pneumoniae P1 protein-binding protein, its preparation method, and its uses. The Mycoplasma pneumoniae P1 protein-binding protein includes an immunoglobulin single variable domain, wherein the immunoglobulin single variable domain comprises CDR1, CDR2, and CDR3 contained in VHH as shown in any one of SEQ ID NO: 1-8. The bivalent or multivalent nanobodies prepared using this Mycoplasma pneumoniae P1 protein-binding protein can effectively enhance their binding strength to antigens, providing a superior raw material for developing highly sensitive detection and diagnostic applications.

The invention discloses a primer probe set for qualitative detection of mycoplasma pneumoniae, an RAA-CHA detection kit and application, and belongs to the technical field of microbiological detection.The primer probe set comprises an upstream primer, a downstream primer, an upstream probe, a downstream probe and a downstream probe, the nucleotide sequence of the upstream primer is as shown in SEQ ID NO: 1, and the nucleotide sequence of the downstream primer is as shown in SEQ ID NO: 2; the nucleotide sequence of the downstream primer is as shown in SEQ ID NO: 2, and the 5'end of the downstream primer is modified by phosphorylation; the nucleotide sequence of the H1 hairpin probe is as shown in SEQ ID NO: 3, and two ends of the H1 hairpin probe are respectively modified with a fluorescence reporter group and a fluorescence quenching group; the nucleotide sequence of the H2 hairpin probe is as shown in SEQ ID NO: 4. According to the application, efficient, accurate and sensitive qualitative detection of the mycoplasma pneumoniae gene is realized through effective amplification and specific recognition of the mycoplasma pneumoniae gene.

The application belongs to the field of intelligent medical treatment, and particularly relates to a method for constructing a refractory mycoplasma pneumoniae pneumonia prediction model and equipment. The method comprises the following steps: acquiring a baseline time data set and a follow-up time label of a mycoplasma pneumoniae patient; inputting the image data set into a lung blood vessel segmentation model to obtain a lung blood vessel network, obtaining a total lung blood vessel volume based on the lung blood vessel network, and obtaining a blood vessel volume of different cross-sectional areas based on a cross-sectional area of the lung blood vessel network; inputting a ratio of the blood vessel volume of different cross-sectional areas to the total lung blood vessel volume into a machine learning model to obtain a prediction label, and obtaining a refractory mycoplasma pneumoniae pneumonia prediction model after iterative optimization of the machine learning model based on a difference between the prediction label and the follow-up time label. The application segments a lung CT image to obtain a blood vessel network diagram, quantitatively obtains a percentage of a blood vessel volume of different thicknesses in a total lung blood vessel volume, and constructs a refractory mycoplasma pneumoniae pneumonia prediction model based on an innovative feature extraction method.

The invention provides a nano antibody specifically bound with mycoplasma pneumoniae P1 protein or an antigen binding fragment thereof, which can specifically recognize and bind the mycoplasma pneumoniae P1 protein and has better affinity with the mycoplasma pneumoniae P1 protein. The mycoplasma pneumoniae P1 protein can be used for preparing products for diagnosing, preventing and / or treating mycoplasma pneumoniae P1 protein infection or diseases caused by the mycoplasma pneumoniae P1 protein infection, or detecting the existence or the level of the mycoplasma pneumoniae P1 protein in a sample.

The application relates to the technical field of detection of mycoplasma pneumoniae, in particular to a rapid detection pen for mycoplasma pneumoniae antibodies, which comprises a box body, one end of the box body is fixedly connected with a drawing cylinder, the other end of the box body is provided with a negative pressure device, a connecting hole is formed through the box body, the two ends of the connecting hole are respectively communicated with the negative pressure device and the drawing cylinder, an installation groove is formed in the box body, a through hole is formed through the installation groove and the connecting hole, a filter layer is fixedly installed in the installation groove, and a detection test paper is fixedly connected to the upper surface of the filter layer. In use, the nasal cavity under different conditions can be sampled by switching modes, and the nasal mucus is drawn into the connecting hole and flows into the installation groove by pressing the negative pressure device, the filter layer removes impurities, optimizes sample concentration, protects target substances and reduces interference, and then flows to the detection test paper for detection, so that the equipment is simple and convenient to use and is convenient to use at home.

Provided is an immunogenic Mycoplasma pneumoniae polypeptide epitope, which is a polypeptide epitope derived from the key adhesion proteins P1, P30, P40 / 90 and P116 of type I and type II Mycoplasma pneumoniae, and / or the CARDS toxin thereof. Further provided is a recombinant protein, which contains one or more of the immunogenic Mycoplasma pneumoniae polypeptide epitopes. Further provided is a vaccine for preventing Mycoplasma pneumonia infections, which vaccine provides protection against at least one of type I and / or type II Mycoplasma pneumonia infections, and is characterized in that the vaccine contains the recombinant protein and is a vaccine capable of providing safe and effective protection against Mycoplasma pneumonia infections.

The invention belongs to the technical field of kits, and particularly relates to a mycoplasma pneumoniae antigen detection kit which comprises a test tube rack, a plurality of first grooves are formed in the test tube rack and used for containing reagent tubes and a box body, the box body is used for containing the test tube rack and a pressurization assembly, and the pressurization assembly is arranged in the test tube rack and used for containing the reagent tubes. By rotating an external threaded column, the external threaded column enters an internal threaded channel, gas in the internal threaded channel is extruded, air pressure is applied to the surface of a piston rod through an air channel and a second groove in sequence, and then the reagent tube is extruded and fixed. At the moment, the piston rod is ejected out of the second groove by the air pressure, enters the first groove and simultaneously contacts with the reagent tube, the air pressure is continuously increased, the air pressure applied to the piston rod enables the piston rod to extrude and fix the reagent tube, collision of the reagent tube in the first groove is prevented, noise is avoided, and damage of the reagent tube caused by collision is reduced.

The application discloses a detection system of mycoplasma pneumoniae. According to the P1 gene of mycoplasma pneumoniae, RPA primers and crRNA required by a CRISPR system with LbCas12a as a core enzyme are designed and screened, an RPA system and a CRISPR system are established, the influence of each component of the CRISPR-LbCas12a system on the RPA system reaction and the influence of each component of the RPA system on each component of the CRISPR-LbCas12a system are tested, components are optimized according to the test results, and an RPA-LbCas12a integrated system capable of detecting a conserved fragment on the P1 gene of mycoplasma pneumoniae is established. The RPA-LbCas12a integrated system established in the application has high sensitivity, high detection efficiency, reduces nucleic acid pollution, and has a wide application prospect.

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.

The present invention discloses an evaluation system and method for improving the participation of Mycoplasma pneumoniae detection by relay detection, which belongs to the field of data analysis. The system includes: a data collection module for collecting behavioral data of a research population; a data preprocessing module for performing data cleaning, integration and preprocessing on the behavioral data; a model training module for obtaining a participation evaluation model; an evaluation import module for obtaining the output results of the participation evaluation model and importing them into the evaluation database of researchers for clustering and segmentation; and a task allocation module for adjusting and allocating detection tasks according to the evaluation and segmentation results of researchers. This application solves the technical problem of the lack of effective methods in the prior art for evaluating and intelligently improving the participation of Mycoplasma pneumoniae detection, and achieves the technical effect of quantitatively evaluating the participation of researchers through a data-driven machine learning model, and optimizing task allocation accordingly, thereby improving the overall efficiency and quality of Mycoplasma pneumoniae detection.

The invention relates to the technical field of mycoplasma pneumoniae detection, in particular to a mycoplasma pneumoniae antibody rapid detection pen which comprises a box body, one end of the box body is fixedly connected with an extraction cylinder, the other end of the box body is provided with negative pressure equipment, a connecting hole is formed in the box body in a penetrating mode, and the negative pressure equipment is connected with the extraction cylinder. A connecting hole is formed in the box body, the two ends of the connecting hole are communicated with negative pressure equipment and the extraction cylinder respectively, a mounting groove is formed in the box body, a through hole is formed between the mounting groove and the connecting hole in a penetrating mode, a filter layer is fixedly mounted in the mounting groove, and detection test paper is fixedly connected to the upper surface of the filter layer. Nasal cavities under different conditions can be sampled by changing modes, the negative pressure equipment is pressed to suck snot into the connecting hole and flow into the mounting groove, the filter layer removes impurities, optimizes sample concentration, protects target substances and reduces interference, and then the snot flows to the detection test paper to be detected. The equipment is simple and convenient to use and can be conveniently used at home.

This invention relates to the field of biomedical technology, specifically disclosing a three-component recombinant subunit vaccine composition for Mycoplasma pneumoniae and its application. The vaccine composition comprises key antigenic proteins of Mycoplasma pneumoniae (MP): P1 protein, P40 / 90 protein, and an E132A attenuated mutant of community-acquired respiratory distress syndrome (CARDS) toxin (CARDS-Mut), optionally combined with Al(OH)3 single adjuvant or Al(OH)3+CPG double adjuvant. The recombinant protein was successfully purified using a prokaryotic expression system, and validated in a mouse model. The vaccine induces high-titer specific IgG antibodies, significantly alleviates lung tissue pathological damage, reduces bacterial load, and inhibits inflammatory factor secretion after challenge, with good safety. This invention solves the problem of high toxicity of CARDS toxin, providing an efficient and safe vaccine strategy for the prevention and control of MP infection.

A kit of detecting Mycoplasma pneumoniae is provided, and the kit comprises a combination of primers and probe consisting of a forward primer, a reverse primer and a probe. The forward primer is selected from the group consisting of SEQ ID NO: 2-3 and complementary sequences thereof, the reverse primer is selected from the group consisting of SEQ ID NO: 6-8 and complementary sequences thereof, and the probe is SEQ ID NO: 9 or a complementary sequence thereof. A method for detecting Mycoplasma pneumoniae using the above kit is also provided.

The invention is applicable to the technical field of biological detection, and provides a primer probe group, system and method for simultaneously detecting mycoplasma pneumoniae and chlamydia pneumoniae, specific primers and probes are designed aiming at conserved regions of 16S rRNA gene of MP and MOMP gene of CP, the two probes are respectively marked with FAM and ROX fluorophores, and the specific primers and probes can be used for simultaneously detecting mycoplasma pneumoniae and chlamydia pneumoniae. Parallel detection of MP and CP nucleic acids can be realized in a single reaction tube. The detection system provided by the invention comprises the PCR enhancer, so that the detection sensitivity and the amplification efficiency are effectively improved. The method further comprises an optimized nucleic acid extraction step and a reaction procedure, has the advantages of being easy and convenient to operate, rapid in detection, high in sensitivity and high in specificity and is suitable for early-stage rapid diagnosis of clinical respiratory tract samples.

The present invention provides a recombinant human Mycoplasma pneumoniae adhesion protein vaccine comprising human Mycoplasma pneumoniae adhesion proteins P1, P30, P40 / 90, and P116 fusion proteins C1, C1-G4S, C2, and C2-G4S. Upon immunization of mice with the recombinant human Mycoplasma pneumoniae adhesion protein vaccine, purified and expressed in Escherichia coli, the vaccine induces a strong immune response in vivo, reduces cytokine production, and reduces Mycoplasma pneumoniae colonization in lung tissue. The vaccine can be used for the prevention, detection, or treatment of Mycoplasma pneumoniae infection.

The invention provides a mycoplasma pneumoniae vaccine and application thereof. Specifically, the invention relates to a mycoplasma pneumoniae protein, a vaccine composition composed of the mycoplasma pneumoniae protein, a preparation method and medical application of the mycoplasma pneumoniae protein, and the vaccine composition has a good protection effect on mycoplasma pneumoniae infection, and can remarkably relieve weight loss, reduce lung pathogen load and reduce lung pathological score. The mycoplasma pneumoniae vaccine has extremely high application value in mycoplasma pneumoniae vaccine development.

The invention provides a mycoplasma pneumoniae polypeptide epitope with immunogenicity. The mycoplasma pneumoniae polypeptide epitope is a polypeptide epitope which is from mycoplasma pneumoniae type I and type II and adheres to key protein P1, P30, P40 / 90, P116 and / or CARDS toxin. The invention also provides a recombinant protein which contains more than one mycoplasma pneumoniae polypeptide epitope with immunogenicity. The invention further provides a vaccine for preventing mycoplasma pneumoniae infection, the vaccine provides protection against infection of at least one of mycoplasma pneumoniae type I and / or type II, and the vaccine is characterized by comprising the recombinant protein. According to the present invention, it is possible to provide a safe and effective vaccine for preventing mycoplasma pneumoniae infection.

The invention discloses a Cas12a allosteric regulation-based portable hypersensitive detection kit for mycoplasma pneumoniae and application, the kit comprises an RAA amplification system, a Cas12a recognition system and a free nucleic acid probe system, when a mycoplasma pneumoniae target exists, an amplification product firstly activates a classical Cas12a / crRNA compound, Cas12a generates conformational change, and the Cas12a recognition system and the free nucleic acid probe system generate allosteric regulation; then the free split-CrRNA probe competitively binds and continuously activates Cas12a, multiple rounds of reporter probe shearing are achieved, so that a detection signal is greatly amplified, and the sheared reporter probe can be dropwise added to detection test paper due to the fact that the sheared reporter probe carries a Biotin group to achieve portable visual detection. The method has the advantages of extremely high sensitivity, simplicity and convenience in operation, instant on-site detection and the like, and is particularly suitable for primary medical treatment and rapid on-site screening.

The invention relates to the technical field of gene detection, and provides a mycoplasma pneumoniae nucleic acid detection quality control product based on a recombinant artificial chromosome and application of the mycoplasma pneumoniae nucleic acid detection quality control product. Aiming at the problems of difficulty in integration of multi-target full-length genes, accurate drug-resistant mutation, insufficient stability and the like of an existing quality control product, a recombinant artificial chromosome carrying full-length sequences of mycoplasma pneumoniae key target genes (16S rRNA, 23S rRNA, a P1 adhesion protein gene and a community-acquired respiratory distress syndrome toxin gene toxin) is constructed; important clinical drug-resistant mutation sites (A2063G, A2064G, C2617T and A2067G) are accurately introduced into the 23S rRNA gene, and the 23S rRNA gene is stably integrated into escherichia coli engineering bacteria by using a bacterial artificial chromosome system. The quality control product exists in a low-copy form, has the advantages of multi-target coverage, accurate simulation of drug-resistant mutation, high genetic stability and no biological safety risk, and is suitable for performance analysis verification and quality control of a detection kit.

The invention discloses a centrifugal micro-fluidic device for mycoplasma pneumoniae detection, comprising: a micro-fluidic chip comprising a chip body and at least one detection unit; the chip body is provided with a rotation center; the detection unit comprises a washing liquid chamber; the device comprises a washing liquid chamber, a blasting valve, a sample and magnetic bead liquid chamber, a siphon valve, an eluent chamber and a pneumatic chamber, one end of the blasting valve is communicated with the washing liquid chamber, and the eluent chamber is respectively communicated with one end of the pneumatic chamber and one end of the siphon valve; a magnetic attraction piece is arranged in the mixing chamber, and the other end of the explosion valve, the sample and magnetic bead liquid chamber and the other end of the siphon valve are all communicated with the mixing chamber; the flow dividing valve is communicated with the mixing chamber; the first waste chamber is communicated with the diverter valve; the reaction unit comprises at least one metering chamber, and the at least one reaction chamber and the at least one metering chamber are communicated with the diverter valve. The invention also discloses a use method of the centrifugal microfluidic device. According to the invention, nucleic acid extraction, amplification and detection are integrated, the detection efficiency is high, and the detection cost is low.

The invention relates to the field of mycoplasma pneumoniae detection, in particular to a primer, a probe and a kit for detecting a drug-resistant gene of mycoplasma pneumoniae. The kit is simple to operate, short in detection time and accurate and reliable in detection result; meanwhile, the method can also be combined with an isothermal amplification technology to improve the detection sensitivity, is suitable for being used in grassroots units and battlefields with limited environmental conditions, and has a huge application prospect.

The invention belongs to the field of intelligent medical treatment, and particularly relates to a construction method and equipment of an intractable mycoplasma pneumonia prediction model. The method comprises the following steps: acquiring a data set of baseline time and a follow-up time label of a mycoplasma pneumoniae patient; inputting the image data set into a pulmonary blood vessel segmentation model to obtain a pulmonary blood vessel network, obtaining a total pulmonary blood vessel volume based on the pulmonary blood vessel network, and obtaining blood vessel volumes of different cross section areas based on the blood vessel cross section areas in the pulmonary blood vessel network; and inputting the ratio of the blood vessel volume of different cross section areas to the total pulmonary blood vessel volume into a machine learning model to obtain a prediction label, and iteratively optimizing the machine learning model based on the difference between the prediction label and the follow-up time label to obtain an intractable mycoplasma pneumonia prediction model. According to the method, the lung CT image is segmented to obtain the blood vessel network graph, the percentage of the volume of blood vessels with different thicknesses in the total lung blood vessel volume is obtained through quantification, and the refractory pneumonia prediction model is constructed based on an innovative feature extraction mode.

The application provides a method for establishing and optimizing a mycoplasma pneumoniae detection system based on ERA-CRISPR / Cas12a. The application provides a method for detecting whether mycoplasma pneumoniae exists in a sample and a corresponding kit. The method and the kit of the application can quickly, economically, highly sensitively and highly specifically detect mycoplasma pneumoniae.

The invention provides a composition with sore-throat relieving, cough relieving and antiviral effects as well as a preparation method and application thereof, and belongs to the technical field of pharmaceutical compositions. The composition provided by the invention comprises 11-20 parts of fermented soybean, 22-26 parts of medicated leaven, 1-3 parts of blackberry lily, 2-4 parts of platycodon grandiflorum, 5-8 parts of perilla leaves, 32-35 parts of fructus aurantii, 1-3 parts of coptis chinensis, 4-8 parts of trichosanthes kirilowii maxim, 5-9 parts of loquat leaves, 2-4 parts of scutellaria baicalensis, 1-2 parts of cape jasmine, 5-9 parts of mangnolia officinalis, 4-8 parts of radix bupleuri, 12-16 parts of rhizoma pinelliae preparata, 5-8 parts of radix curcumae, 12-18 parts of liquorice, 4-6 parts of fresh ginger, 32-36 parts of Chinese dates and 4-8 parts of poria cocos. On the basis of adjusting the monarch, minister, assistant and guide relationship, the composition disclosed by the invention is combined with the improvement of the preparation method, so that efficient extraction of components, synergistic enhancement of the drug effect and process optimization are realized, and finally the remarkable technical effects of relieving sore throat and cough, and resisting viruses and mycoplasma pneumoniae are achieved.

The utility model discloses a kit for detecting mycoplasma pneumoniae, which belongs to the technical field of biochips and diagnostic reagents, and comprises a positive quality control reagent, a negative quality control reagent and a detection chip, the detection chip comprises an upper shell and a lower shell, the upper shell is connected with the lower shell in a clamping manner, the upper shell is provided with three sample introduction windows, and the lower shell is provided with three sample introduction openings. The upper shell and the lower shell are provided with three sample introduction windows, the three sample introduction windows are respectively a negative control port, a positive control port and a detection port, and a reaction film is arranged between the upper shell and the lower shell; a negative control area is arranged at the position, opposite to the negative control opening, of the reaction film, a positive control area is arranged at the position opposite to the positive control opening, and a detection area is arranged at the position opposite to the detection opening. The kit disclosed by the utility model can accurately verify whether the quality of the detection chip is qualified or not and whether manual operation is correct or not, so that the quality control of a product is enhanced, and the accuracy of a detection result is improved.

This invention discloses a Mycoplasma pneumoniae antibody detection kit based on immunoturbidimetry, belonging to the field of clinical immunology testing. It includes solutions R1 and R2. Solution R1 is a dispersion of double-coupled latex microspheres coated with natural Mycoplasma pneumoniae antigens, and solution R2 is a sample diluent containing an enhancer. The test samples can be serum, plasma, or whole blood samples, and it is applicable to biochemical analyzers, protein analyzers, or combined blood cell and immunoturbidimetric analyzers. The kit prepared by this invention can be used on various instruments, has a wide range of detection targets, and can be coupled with other instruments to detect various clinically valuable blood indicators, further shortening the testing time. It solves the problems of poor accuracy, low efficiency, and limited application scenarios caused by the subjective interpretation of results, long processing time, and limited applicability of blood samples in existing technologies. It can effectively simplify the testing process and improve diagnostic efficiency.

This invention provides a subunit vaccine containing Mycoplasma pneumoniae HP14 / 30 fusion antigen and a mucosal adjuvant. The subunit vaccine comprises the Mycoplasma pneumoniae HP14 / 30 fusion antigen and a mucosal adjuvant in a mass ratio of 1-5:1. The HP14 / 30 fusion antigen is a recombinant protein co-expressed by chimeric fragments of the P1 and P30 proteins, with the amino acid sequence shown in SEQ ID NO:1. This vaccine is administered via mucosal routes such as nasal drops or oral inhalation. It efficiently induces respiratory mucosal-specific IgA and humoral IgG responses, significantly reduces pathogen load in the lungs, alleviates pathological damage to lung tissue, and does not induce enhanced respiratory disease (ERD). Its safety and protective efficacy are significantly superior to traditional inactivated vaccines and intramuscular vaccines. This invention fills the technological gap in human Mycoplasma pneumoniae subunit vaccines, is suitable for the prevention of Mycoplasma pneumoniae infection in children and adults, and has significant clinical application value.

The invention discloses a detection system of mycoplasma pneumoniae. According to the application, an RPA primer and crRNA required by a CRISPR system taking LbCas12a as a core enzyme are designed and screened according to a mycoplasma pneumoniae P1 gene, an RPA system and the CRISPR system are established, then the influence of each component of the CRISPR-LbCas12a system on the reaction of the RPA system and the influence of each component of the RPA system on each component of the CRISPR-LbCas12a system are tested, the components are optimized according to the test results, and the detection result is accurate. According to the invention, the RPA-LbCas12a integrated system capable of detecting conserved fragments on the mycoplasma pneumoniae P1 gene is established, and the RPA-LbCas12a integrated system established by the invention has the advantages of high sensitivity, high detection efficiency, reduction of nucleic acid pollution and wide application prospect.

Provided are monoclonal antibodies and antigen-binding fragments that specifically bind the CARDS toxin or the P1 adhesin of Mycoplasma pneumoniae. The antibodies enable rapid, sensitive detection of M. pneumoniae infection and provide therapeutic neutralization of toxin activity or inhibition of bacterial adherence. Also provided are nucleic acids, vectors, host cells, pharmaceutical compositions, diagnostic methods, and kits using these antibodies.