204 results about "Mycobacterium tuberculosis complex" patented technology

An assay method and kit is disclosed for detecting the presence of at least one predesignated, target antibody to a mycobacterium in a sample selected from one or more patient bodily fluids. The method comprises the following steps: (a) contacting the sample of one or more patient bodily fluids with at least one mycobacterium antigen on a lateral-flow assay membrane to bind to the target antibody in the sample; (b) previously, simultaneously or subsequently to step (a), binding the at least one mycobacterium antigen with a conjugated label producing a detectable signal; and (c) detecting the signal whereby the presence of the target antibody is determined in the sample by the intensity or presence of the signal. The method can further comprise the step of evaluating immunization status of the patient from whom the sample came by comparing the signal or lack thereof with immunizations previously received by the patient and in comparison to a known standard control. In a preferred embodiment, the mycobacterium antigen specifically binds to Mycobacterium tuberculosis specific antibodies. Preferably, the immunoassay of the present invention comprises a lateral-flow assay comprising a membrane, a conjugated label pad, and at least one mycobacterium antigen bound to the membrane. In a preferred embodiment, the at least one mycobacterium antigen is selected from the group consisting of 38 kDa and 16 kDa antigens.

The present invention relates to compositions and fusion proteins containing at least two Mycobacterium sp. antigens, and nucleic acids encoding such compositions and fusion proteins. The compositions of the invention increase serological sensitivity of sera from individuals infected with tuberculosis, and methods for their use in the diagnosis, treatment, and prevention of tuberculosis infection.

The present invention relates to compositions and fusion proteins containing at least two Mycobacterium sp. antigens, and nucleic acids encoding such compositions and fusion proteins. The compositions of the invention increase serological sensitivity of sera from individuals infected with tuberculosis, and methods for their use in the diagnosis, treatment, and prevention of tuberculosis infection.

The invention belongs to the field of biomedicine examination, and particularly relates to a kit and a method for detecting mycobacterium tuberculosis infection and application. The invention discloses a novel mycobacterium tuberculosis detection reagent by screening specific T cell epitope of mycobacterium tuberculosis, wherein the reagent contains polypeptide or analog thereof represented by SEQ ID No.1-10. The method detects cell factors released from T cells by using single or more SEQ ID No.1-10 polypeptides to contact the T cells of mycobacterium tuberculosis infected individuals. The method can effectively detect active tuberculosis or latent tuberculosis infection, and is free from disturbance of Bacilli Calmette Guerin (BCG) inoculation vaccines. The invention also discloses a diagnostic kit and other application based on the polypeptide and the method. Compared with the gamma interferon release experiments in the prior art, the method can obviously improve the detection rate without reducing the specificity and has high clinical application value.

The present invention relates to molecular biology, microbiology, and medicine and provides the method for detection of Mycobacterium tuberculosis complex with simultaneous evaluation of sensitivity of the strains to rifampicin and isoniazid in clinical sample on differentiating biochip. The method is based on two-stage multiplex PCR to obtain fluorescent DNA fragments followed by hybridization of these fragments on microarray containing the set of specific discriminating oligonucleotides. The determination of the resistance of Mycobacterium tuberculosis to rifampicin and isoniazid is carried out by evaluation of point nucleotide substitutions in DNA of microorganism. The present invention allows conduct analysis directly in clinical sample, to evaluate a number of mutations simultaneously, to decrease the cost price of analysis, and to reduce the time of its conducting. The present invention also relates to set of primers, biochip, and set of oligonucleotide probes used in realization of the method.

The present invention relates to fusion proteins containing at least two Mycobacterium tuberculosis antigens. In particular, it relates to bi-fusion proteins which contain two individual M. tuberculosis antigens, tri-fusion proteins which contain three M. tuberculosis antigens, tetra-fusion proteins which contain four M. tuberculosis antigens, and penta-fusion proteins which contain five M. tuberculosis antigens, and methods for their use in the diagnosis, treatment and prevention of tuberculosis infection.

Contemplated compositions, devices, and methods are drawn to various antigens from the pathogen M. tuberculosis and their use in vaccines, therapeutic agents, and various diagnostic tests. In particularly preferred aspects, the antigens are immunodominant and have quantified and known relative reactivities with respect to sera of a population infected with the pathogen, and/or have a known association with a disease parameter.

The invention belongs to the technical field of nucleic acid detection, and relates to a mycobacterium tuberculosis complex detection kit and method based on a CRISPR-Cas12a system. According to the kit, the detection sensitivity is improved by adopting a recombinase polymerase amplification technology, CRISPR-Cas12a is used for specifically targeting a mycobacterium tuberculosis complex target sequence and then activating the bypass cutting activity of Cas12a, and a mycobacterium tuberculosis complex can be sensitively and specifically detected from sputum. The kit and method have the advantages of being non-invasive, capable of detecting the mycobacterium tuberculosis complex frequently and repeatedly, high in detection speed and the like. Compared with an existing PCR detection method,the method based on the CRISPR-Cas12a system does not need a thermal cycler with heating and cooling functions, can detect the mycobacterium tuberculosis complex in sputum through fluorescence reading, has the advantages of being low in cost, easy and convenient to operate, high in sensitivity, good in specificity and the like and is suitable for clinical large-scale application.

The invention discloses a method for quickly detecting mycobacterium tuberculosis, which comprises the following steps: 1, infecting phage with the mycobacterium tuberculosis; 2, killing the phage notinfecting outside the mycobacterium tuberculosis; 3, amplifying the infecting phage; and 4, after labeling gold with antiphage antibody, adopting quick immunochromatography to detect whether the phage exists or not so as to judge whether the mycobacterium tuberculosis exists in a sample. In addition, the invention also discloses a corresponding kit, which comprises sample treating fluid, proliferous liquid, a killing agent, mycobacterium smegmatis, mycobacteriophages and a phage colloidal gold method immunity-chromatography test pen. The method and the kit can quickly and accurately detect the mycobacterium tuberculosis so as to meet the requirement of quickly diagnosing pulmonary tuberculosis and timely medicate the patient.

The invention relates to a tuberculosis antibody multiple antigen ELISA detection kit and a preparation method thereof, which pertains to the field of tuberculosis medical immunology diagnostic techniques and mainly uses detection antigen, enzyme-linked antihuman IgG antibodies, substrates, positive control serum of tuberculosis patients, control serum of normal person, calf serum and polystyrene microplates to form the kit, wherein, the detection antigen adopts the mycobacterium tuberculosis complex strains of lipid Arabian mannose (LAM), 38kD and 16kD to be combined with arbitrary one or more than one mycobacterium tuberculosis recombinant proteins in the recombinant proteins of MPT63, MTB48 and CFP10-ESAT6. The mycobacterium tuberculosis has high sensitivity, strong specificity and complementarity, can be used for detecting specific antitubercular antibodies in such body fluid samples as serum, hydrothorax and the like, and assisting the diagnosis and differential diagnosis of tuberculosis.

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.

The present invention provides a method for identifying Mycobacterium tuberculosis and non-tuberculosis Mycobacterium (MOTT), and for the determination of drug susceptibility of M. tuberculosis based on detection of mutations in the rpoB gene.

The invention discloses a method and a kit for detecting the streptomycin medicine resistant mutation of Mycobacterium tuberculosis. The invention relates to medicine resistant mutation detection technique and provides a method for detecting streptomycin medicine resistant mutation of Mycobacterium tuberculosis, which effectively improves sensitivity and specificity, and is simple and convenient in operation and short in period. The method comprises: designing primers and probes according to the complete sequence of the Mycobacterium tuberculosis and the gene sequences of the genomes rpsL and rrs of the Mycobacterium tuberculosis; extracting the DNA of a sample of the Mycobacterium tuberculosis; constructing a polymerase chain reaction (PCR) reaction system; and performing PCR amplification and analysis on a fusion curve. In the method, experiments are performed in two tubes respectively by using the specific primers and probes, and the amplification of the nucleic acid fragment of a target nucleotide sequence and subsequent analysis on the fusion curve are realized by using heat-resistance DNA polymerase, four kinds of nucleotide monomers and other components and by using real-time PCR technique. A fluorescent PCR fusion curve method with high specificity can quickly and accurately detects common medicine-resistance mutation of Mycobacterium tuberculosis and is expected to be directly used for medicine-resistance detection of a clinic Mycobacterium tuberculosis sample.

This invention relates to a method for detecting Mycobacterium tuberculosis by utilizing gamete technique. This invention is aimed to solving the problems of low discovery rate and low curative rate of tuberculosis. The method comprises: utilizing gamete technique, performing bacteria culture or bioengineering method to obtain all kinds of targets of Mycobacterium tuberculosis, obtaining highly compatible specific gametes of the targets by gamete technique screening, and converting the gametes into report gametes for rapid and precise detection of corresponding targets. The method can further combine highly compatible gametes with pathogenetic factors to defunctionalize them, or develop new drugs by finding structural analogues of highly compatible gametes, thus can provides basis for diagnosis and clinical therapy of Mycobacterium tuberculosis.

The invention belongs to the field of biomedicine examination, and relates to an MTB (Mycobacterium Tuberculosis) infection diagnosis kit. The MTB infection diagnosis kit provided by the invention is prepared from the following components: an antigen stimulant, a pre-coated ELISPOT (Enzyme-Linked Immunospot Assay) plate of a capture antibody, an insulin-like growth factor, a detection antibody, HRP (Horse Radish Peroxidase)-labeled streptavidin, a 3-amino-9-ethyl carbazole developing solution, antibody diluent and a positive control stimulant; the antigen stimulant is selected from one or multiple polypeptides in sequences as shown in SEQ ID NO.1 to 12. The MTB infection diagnosis kit provided by the invention is high in sensitivity and specificity and stable in property; meanwhile, when the MTB infection diagnosis kit provided by the invention is applied to in-vitro detection on MTB, the detection time can be saved, the detection steps can be simplified, and the detection efficiency can be increased.

The invention provides an antigen stimulant for detecting mycobacterium tuberculosis infection, and a kit comprising the antigen stimulant. The invention also provides an application of the antigen stimulant in reagents for detecting mycobacterium tuberculosis infection. The antigen stimulant comprises at least one polypeptide or analogues thereof in polypeptides shown as the sequences 1-11 in a sequence table, wherein the polypeptides respectively come from tuberculosis specific antigen polypeptides ESAT-6 and tuberculosis specific antigen polypeptides CFP-10. According to the antigen stimulant provided by the invention, peripheral blood T lymphocytes of tuberculosis infection patients can be effectively stimulated to generate IFN-gamma, so that the tuberculosis infection can be diagnosed at high sensitivity and high specificity, and the influence on BCC inoculation or other underlying diseases can be avoided.

The invention utilizes a genetic engineering method to express in vitro the desA protein of mycobacterium tuberculosis and produces monoclonal antibodies by using the protein. A tuberculosis specific antigen rapid diagnosis reagent kit is produced by utilizing enzyme linked immunization, chemiluminescence, colloidal gold test strip method and other diagnostics methods so as to detect MTB quickly with low cost. The technique has wide application value in clinical diagnosis.

The invention relates to a diagnostic kit for identification of mycobacterium strains and detection of drug resistance and a preparation method thereof. The kit can quickly and accurately distinguish between the identification of mycobacterium strains and the detection of drug resistance in large quantities by using an oligonucleotide chip. Point mutations in the rpoB gene of Mycobacterium tuberculosis associated with drug resistance. The diagnostic kit for the identification of mycobacterium strains and the detection of drug resistance of the present invention contains an oligonucleotide chip, and the chip contains a complex composed of mycobacterium tuberculosis mycobacterium strain identification probes and drug resistance detection probes. Probe, mycobacterium tuberculosis rpoB gene, and fluorescent material, wherein the fluorescent material contains biotin-binding protein, which is used as the label of biotin and the corresponding probe to detect the hybridization of the sample amplification product.

The invention provides a kit for diagnosing mycobacterium tuberculosis infection based on tuberculosis specificity IL-31 detection. The kit comprises an IL-31 capture antibody, an IL-31 detection antibody, tuberculosis specific antigen polypeptide, a diluent, a standard substance, a developing solution, a stop solution, a buffer solution, a solid carrier and a micro-plate sealer. The kit is used for detecting the IL-31 expression quantity in plasma, can be used for tuberculosis diagnosis, and is relatively high in sensitivity and accuracy, and fast to operate, so that the diagnosing efficiency of tuberculosis can be improved.

The invention discloses immune colloidal gold test paper for detecting mycobacterium bovis antibody and a preparation method thereof. Staphylococal Protein A (SPA) marks colloidal gold to prepare a gold colloidal pad of the immune colloidal gold test paper. The proteins of CFP10 and MPT64 are immunogenic proteins with high specificity to virulent mycobacterium tuberculosis. The genes of CFP10 andMPT64 are cloned from the genome of the mycobacterium tuberculosis, and are connected to pET-28a to construct two prokaryotic expression recombinant plasmids; the two plasmids are converted into Escherichia coli to express the proteins of CFP10 and MPT64; after the proteins are purified, the mixed proteins are taken as antigens to coat a nitrocellulose membrane to form a detection line; and the detection line is equipped with the gold colloidal pad to form the immune colloidal gold test paper. The test paper can distinguish that human bodies and animals are inoculated with BCG or subjected towild virus infection, can be used for detecting the mycobacterium tuberculosis antibody in animal serum, has the characteristics of strong specificity, high sensitivity, good stability, convenience and quickness, and has significance and actual application value for monitoring, diagnosing, purifying and controlling the mycobacterium tuberculosis.

The invention discloses a biomarker for diagnosing mycobacterium tuberculosis infection. The biomarker comprises IL-2 and IL-10. The invention proves that cell factors IL-2 and IL-10 can serve as markers for tuberculosis infection diagnosis for the first time, the tuberculosis infection can be diagnosed by measuring the concentration of the IL-2 and the IL-10 after peripheral blood monouclear cells are stimulated by tuberculosis antigen, and active tuberculosis and latent tuberculosis infection are further distinguished. A tuberculosis infection blood detection reagent and a kit which are prepared on the basis of the two cell factors provide new technological basis for quick diagnosis of the active tuberculosis, have the advantages of high sensitivity, high specificity and the like, are quick in operation, reasonable and practical, and can obviously improve the diagnosis efficiency of the active tuberculosis.