TWM420694U - Hybrid strip for rapid detection of mycobacterium tuberculosis - Google Patents

Description

M420694 V. New description: [New technical field] This is a rapid test strip. [Prior Art] The common laboratory diagnostic methods for Mycobacterium tuberculosis (MTBC) are mainly acid-resistant staining and bacterial culture. Microscopic observation of stained acid-fast bacilli is a method for rapid screening of clinical specimens, but its sensitivity is low, and it is indistinguishable from distinguishing Mycobacterium tuberculosis from patients with immunodeficiency (eg non-sister nuclear) Mycobacterial infection (NTM). Although bacterial culture has good sensitivity and specific I1 production, it usually takes at least 10 days to 6 weeks to detect a positive sample, and then it needs to be judged as MTBC or NTM. The new generation of detection methods utilizes nucleic acid amplification technology to provide very fast and specific detection results directly for MTBc. In theory, in this family, these new generation nucleic acid amplification techniques can measure lower molecules. The gene sequence 'but the general city acid test · Yan in the clinical examination _ shows: the reaction is often subject to nuclear Wei and the scales should inhibit the interference of the machine, the actual sensitivity is often lower than the bacterial culture. The R&D-shopping technology stripassay can be used to eliminate JMTBC and simultaneously report whether there is interference with the nucleic acid-polymerized plum reaction inhibitor, which can effectively reduce the false negative rate of detection, and Low number of bacteria smear sample detection condition of a female born 〖enhance its sensitivity to detect the measured Mtbc. 3 M420694 [New content] This creation department - miscellaneous branches - bribe _ view, including the bottom layer of the test paper 'here is a multi-layer fiber membrane with water absorption, = section from the lower end of the test paper to the upper Wei order for the sample layer, reactants The ruthenium layer, the measurement layer, and the water absorbing layer, wherein the result detection layer: the control line, and the - system control line. ] Line 1 internal creator of the creation device, _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Reactant release material - a water-absorbing support material of a multi-sonic film, such as glass fiber or its equivalent, coated with colloidal gold or its equivalent on the layer; the result is a multi-material For example, a stone-like fiber membrane or an equivalent of the same; the water-absorbing layer is composed of a water-absorbent material, which can increase the water absorption force of the capillary phenomenon. As a result of the creation, the antibody detection line is coated on the detection layer, and the detection line, the internal control line, and the system control line are sequentially from the lower end to the upper end of the test paper. The detection line contains a special 'f-antibody' which can form a specific binding with an antigen on a nucleic acid polymerase chain reaction (PCR) product with a pure tuberculosis (MTB). The internal ligature contains a specific antibody, which can be combined with The antigen on the pCR product of the internal control group forms a specific binding, and the system control line contains a complex ligand that can be combined with all pCR products to detect whether the test paper is functioning properly. After the sample layer is immersed in the PCR product to be tested, the analyte is passed through the reactant release layer by capillary action and the gold is transferred to the result to detect the binding of the active antibody. 4 M420694 can be stacked and colored. This creation also contains two pairs of primer pairs of tuberculosis (4) and the internal control group. (IV) Progenitor (4) 'Induction of Mycobacterium tuberculosis for each marker biograph _ and = prime (FITQ 'internal position (four) sub-to-label * bio thief ) and Dig0xigenin; the test sample can simultaneously replicate a large number of specific phase genes with luciferin, biotin, and digitalis toxicant via the polymerase chain reaction. It can be directly detected by the inspection paper. [Embodiment] As shown in Fig. 3, the sample to be tested of the present invention needs to be treated by extracting deoxyribozyme nucleic acid 1να and nucleic acid polymerase chain reaction (PCR). The process of extracting deoxyribonucleic acid (DNA) is as follows: - Purification of clinical specimens - Collection of Ba bed specimens ' and storage in a 4 °c refrigerator. 2. Make a permanent mark on each test tube. 3. Take 1 gram of N-acetyl cysteine (NALC) dissolved in 50 ml of 8% sodium hydroxide (NaOH), 50 ml of 5.88% sodium citrate, and 1 ml of a mixture of 2 〇q/() sodium dodecyl sulfate (SDS). 4. Add an equal amount of the mixed solution of step 3 to each tube of the clinical specimen. 5. Shock the test tube for 30 seconds' and turn it over 1 time, then let it stand at room temperature for 15 minutes. 6_ Add the Potassium Buffer (PBS) to the 50 ml mark in the test tube and centrifuge for 15 minutes at 3,8 〇〇 5 M420694 centrifugal force (xg). 7. Remove the sample tube from the centrifuge and open the lid. Slowly remove the supernatant and discard. 8. Add 1 ml of PBS to each tube and dissolve the pellet back. 9. Take the i ml of the sinking material and mix it with 1 ml of secondary water for 2 seconds and centrifuge at 3,800 xg for 15 minutes. 10. Remove the test tube from the centrifuge and open the lid. Slowly remove all supernatant and discard. 11. The precipitate is ready for DNA extraction. If the DNA is not extracted immediately, store the pellet in _7〇. Hey.

- Extract DNA U^^LysisTubemM25_4^^^LysisBuff>er) 1 ' and shake for 1 minute, and let stand at room temperature for 1 minute. 2. Move the LysisTube to the preheated dry bath and continue heating for 20 minutes at 1〇0. 3. Allow the Lysis Tube to cool at room temperature for at least 5 minutes, allow the vapor to evaporate, and then quickly centrifuge to sink the sample in the tube. 4_ Pipette 20 μl of LysisBuffer 2 and 15 μl of 1 mM molar Tris-HCl (pH 8.0) into the Lysis Tube and shake for 1 minute. 5. Centrifuge at 14,000 X g for 2 minutes, collect the DNA lysate and carefully pipet 5 μl of the supernatant (only 5 μl in this assay) into a microcentrifuge tube and discard the remaining solution. If the DNA lysate is not treated immediately, it should be stored at _7〇. Hey. The nucleic acid polymerase chain reaction (PCR) process is as follows: 6

Claims (1)

M420694 VI. Scope of Application: ^ A rapid test strip for Mycobacterium tuberculosis, comprising: a support, a sample layer, a reactant release layer, a result detection layer, and a water absorbing layer, wherein the bottom layer for detecting iron is the aforementioned The layer of the support layer and the surname of the fruit are static, and the sample layer and the reactant release: the Γ and the water absorbing layer are arranged in sequence from the lower end to the upper end of the test paper: the detection layer has a detection line and an internal Control line, and a system control line. 2. For the rapid detection test paper of Mycobacterium tuberculosis described in the first paragraph of the patent application, wherein the branching system consists of an inert hard material that does not absorb water. 3. If you apply for the replacement of the No. 1 tuberculosis branching rod and speed test strip, the sample layer and the water absorption and self-water absorption ο 4. If you apply for a patent, please call the tuberculosis branch to quickly test the test paper. Reaction to Lin _Lf Na body money its _ equal. 5. The rapid detection test paper of Mycobacterium tuberculosis according to the scope of claim 1 of the patent application, wherein the reactant release layer is composed of a water-absorbing material. 6. The rapid detection test paper of Mycobacterium tuberculosis according to claim 5, wherein the water-absorbing support material is glass fiber or an equivalent of the same. 7. For rapid detection test of Mycobacterium tuberculosis as described in item 1 of the patent application, M420694 wherein the result detection layer is composed of a porous material. 8. The tuberculosis branching rod speed detecting test paper according to the seventh aspect of the patent application, wherein the porous material may be a nitrocellulose membrane or an equivalent of the same. 9. For example, the tuberculosis branching rod _ speed test strip according to the patent application item i, wherein the test line contains a specific antibody, and can be linked with a nucleic acid polymerase chain with M. tuberculosis (mtb) (PC·物The anti-(4) is combined with the specificity. 10. As described in the patent scope of item i, the M. tuberculosis rapid test strips, wherein the internal control line contains specific antibodies, can form antigens on the PCR products of the internal control group. Specificity. η. For application, please refer to the tuberculosis branching rod g rapid test strip described in the third paragraph, wherein the system control line contains a complex ligand, which can be combined with all pcR products to detect whether the test strip is working properly. 11