A method and kit for rapidly extracting Mycobacterium tuberculosis nucleic acid from sputum

A technology of Mycobacterium tuberculosis and kits, which is applied in the field of biomedical clinical diagnostic reagents, can solve problems such as purification of contaminated nucleic acids, and achieve the effects of improving the quality of nucleic acid extraction, avoiding biological contamination, and ensuring personal health

Active Publication Date: 2022-05-03
亚能生物技术(深圳)有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

"Nucleic acid extraction composition and its application, reagents and kits containing the nucleic acid extraction composition" (Patent No.: CN202010347016.5) uses traditional silica-based adsorption columns, magnetic beads or glass beads as nucleic acid carriers for purification, after cleaning Different concentrations of salt ions introduced into the solution and strengthening solution may contaminate the subsequent nucleic acid purification risk

Method used

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  • A method and kit for rapidly extracting Mycobacterium tuberculosis nucleic acid from sputum
  • A method and kit for rapidly extracting Mycobacterium tuberculosis nucleic acid from sputum

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Embodiment 1

[0065] Step 1: Add an equal volume of lysate to the sputum sample, and treat at 100°C for 15 minutes to inactivate Mycobacterium tuberculosis in the sputum and release nucleic acid;

[0066] Step 2: Centrifuge at 12,000 rpm for 5 minutes, draw 500 μL of the supernatant into a centrifuge tube, add 300 μL of the resuspended magnetic bead solution containing polyethyleneimine modification, mix well, and let stand for 5 minutes;

[0067] Step 3: Place the centrifuge tube on the magnetic stand and let it stand for 1 min to remove the liquid in the centrifuge tube;

[0068] Step 4: Remove the centrifuge tube, add 500 μL of cleaning solution, shake and mix for 30 seconds;

[0069] Step 5: Place the centrifuge tube on the magnetic stand and let it stand for 1 min to remove the liquid in the centrifuge tube;

[0070] Step 6: Remove the centrifuge tube, add 20-100 μL eluent, mix by pipetting, and let stand for 3 minutes;

[0071] Step 7: Place the centrifuge tube on the magnetic stand...

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Abstract

The invention relates to the field of biomedical clinical diagnostic reagents, in particular to a kit for rapidly extracting Mycobacterium tuberculosis nucleic acid from sputum, comprising a lysate, a magnetic bead suspension, a cleaning solution and an eluent; the lysate includes 3-5mol / L potassium chloride, 3-5mol / L guanidine hydrochloride, 1% potassium ethyl xanthate and 0.2mol / L potassium phosphate buffer; the magnetic bead suspension is a nucleic acid composed of polyethyleneimine-modified magnetic beads Adsorption carrier; the cleaning solution is a hydrochloric acid solution; the eluent is a sodium hydroxide solution; a method for rapidly extracting Mycobacterium tuberculosis nucleic acid in sputum is further proposed, so as to finally realize high-quality and fast extraction of Mycobacterium tuberculosis nucleic acid in sputum Extraction and detection, reduce the cost of extraction, simplify the operation process, avoid the use of volatile organic solvents and high-concentration salt solutions, and reduce the hazards of kit reagents to the health of operators. At the same time, the kit is combined with existing automatic detection technology to achieve Potential for high-throughput, large-scale nucleic acid extraction and detection.

Description

technical field [0001] The invention relates to the field of biomedical clinical diagnostic reagents, in particular to a method and a kit for rapidly extracting Mycobacterium tuberculosis nucleic acid from sputum. Background technique [0002] Mycobacterium tuberculosis (Mycobacterium tuberculosis, referred to as Mycobacterium tuberculosis) is the pathogen that causes tuberculosis. This bacterium can infect the whole body tissues and organs of the human body, mainly the lungs, and poses a major threat to human life and health. With the emergence of anti-tuberculosis drugs and BCG vaccine and the improvement of medical level, the infection of Mycobacterium tuberculosis once dropped sharply. However, due to the abuse of antibiotics, the emergence of drug-resistant strains and the prevalence of AIDS, the prevalence of tuberculosis worldwide tends to worsen. According to World Health Organization estimates, in 2018, there were about 10 million new cases of Mycobacterium tubercu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
CPCC12N15/1013C12N15/1003
Inventor 郭威林杜维任保彦薛良
Owner 亚能生物技术(深圳)有限公司
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