Real-time fluorescent nucleic acid isothermal amplification detection kit for mycobacterium tuberculosis as well as special primer and probe thereof
A technology for the detection of mycobacterium tuberculosis and constant temperature amplification, which is applied in the field of biomedical detection, can solve problems such as mutual interference, interference, and detection sensitivity reduction, and achieve high detection sensitivity
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0058] Example 1: Design of special primers and probes for real-time fluorescent nucleic acid constant temperature amplification detection of Mycobacterium tuberculosis The inventor disclosed the detection sequence according to ZL201110137694.X (its nucleotide sequence is shown in SEQ ID NO: 1 ), carry out primer and probe design according to primer and probe design principle, to carry out real-time fluorescent nucleic acid constant temperature amplification detection to Mycobacterium tuberculosis.
[0059] This embodiment has designed multiple groups of primers and probes altogether, wherein selects following three groups of primers and probes (group 1, group 2 and group 3) to Mycobacterium tuberculosis positive control and negative control (do not contain Mycobacterium tuberculosis nucleic acid system, such as physiological saline) for detection verification, so as to screen out primers and probe sets with good specificity, sensitivity and reproducibility for the detection of...
Embodiment 2
[0084] Embodiment 2: Real-time fluorescent nucleic acid constant temperature amplification detection kit for Mycobacterium tuberculosis
[0085] The kit for the detection of Mycobacterium tuberculosis nucleic acid provided in this example is based on the principle of RNA nucleic acid constant temperature simultaneous amplification detection to detect a gene conservation region of Mycobacterium tuberculosis nucleic acid (16S rRNA, the corresponding DNA sequence is shown in SEQ ID NO: 1 Shown) the kit, on the basis of the primers and probes shown in Group 1 determined in the examples, specifically includes the following components:
[0086] (1) Nucleic acid extraction solution: used for extracting and purifying the Mycobacterium tuberculosis nucleic acid in the sample, containing 250~800mM HEPES, 50~500mg / L magnetic beads (solid phase support), 1~50μM specific capture probe (SEQ ID NO:2), 25~150pmol / mL extraction probe (SEQ ID NO:3);
[0087] (2) Detection solution a: it contai...
Embodiment 3
[0096] Embodiment 3: Real-time fluorescent nucleic acid isothermal amplification method for detecting Mycobacterium tuberculosis
[0097] The method of this embodiment is based on the principle of RNA constant temperature synchronous amplification detection to detect Mycobacterium tuberculosis nucleic acid, which uses the kit provided in Example 2 above to detect whether the sputum sample obtained from the subject contains Mycobacterium tuberculosis nucleic acid, specifically The operation steps are:
[0098] 3.1. Sample processing
[0099] Take 1 mL of sputum sample, add 1 mL of sample pretreatment solution (lithium lauryl sulfate (LLS) 8%, 15 mM Tris-HCl, 0.3 mM EDTA), and vortex for 1 min to obtain a mixed solution. Heat treatment at 90°C for 15 minutes, and then ultrasonic treatment at 300w for 15 minutes to obtain the pretreated sample.
[0100] 3.2. Nucleic acid extraction
[0101] (1) Take 1 mL of the pretreated sample in an EP tube, add 250 μL of nucleic acid extrac...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com