One-tube magnetic bead method for extraction of virus nucleic acid for fluorescent quantitative detection

A fluorescent quantitative detection and viral nucleic acid technology, which is applied in microorganism-based methods, biochemical equipment and methods, and microorganism determination/inspection, etc., can solve cumbersome problems, simplify operation steps, save production costs, and improve detection efficiency. Effect

Inactive Publication Date: 2012-07-25
TIANGEN BIOTECH BEIJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The whole process of this method is divided into five steps, the fourth step requires incubation, elution and separation, which is cumbersome for users who want to quickly extract and detect

Method used

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  • One-tube magnetic bead method for extraction of virus nucleic acid for fluorescent quantitative detection
  • One-tube magnetic bead method for extraction of virus nucleic acid for fluorescent quantitative detection
  • One-tube magnetic bead method for extraction of virus nucleic acid for fluorescent quantitative detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The hepatitis B virus nucleic acid sample (HBV-DNA) was extracted by this method and the magnetic bead elution method, and detected by fluorescence quantitatively.

[0030] 1. Extraction of hepatitis B virus nucleic acid sample (HBV-DNA):

[0031] 1, the inventive method extracts:

[0032] (1) Add 50 microliters of hepatitis B virus serum into a prefabricated strip tube (for fluorescent quantitative detection). The components in the tube are 150 microliters of special virus lysate and 10 microliters of specific magnetic beads, which are thoroughly mixed to achieve the full release of nucleic acids, and then fully combine the magnetic beads with nucleic acids;

[0033] (2) Place the above-mentioned strip tube containing the magnetic beads adsorbed with nucleic acid on the magnetic stand, separate the magnetic particles adsorbed with nucleic acid from the solution by magnetic force, then discard the solution, and take off the strip tube;

[0034] (3) Add 100 microliters...

Embodiment 2

[0060] The method of the invention is used to extract the hepatitis B virus nucleic acid sample (HBV-DNA) after gradient dilution and perform fluorescence quantitative detection.

[0061] 1. Extraction of hepatitis B virus nucleic acid sample (HBV-DNA):

[0062] 1. The inventive method extracts:

[0063] (1) Add 50 microliters of serially diluted hepatitis B virus serum to prefabricated strip tubes (for fluorescent quantitative detection). The components in the tube are 150 microliters of special virus lysate and 10 microliters of specific magnetic beads, which are thoroughly mixed to achieve the full release of nucleic acids, and then fully combine the magnetic beads with nucleic acids;

[0064] (2) Place the above-mentioned strip tube containing the magnetic beads adsorbed with nucleic acid on the magnetic stand, separate the magnetic particles adsorbed with nucleic acid from the solution by magnetic force, then discard the solution, and take off the strip tube;

[0065] (...

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Abstract

The invention relates to a one-tube magnetic bead method for extraction of virus nucleic acid for fluorescent quantitative detection, which belongs to the technical field of biological nucleic acid application and includes: firstly, preparing lysis solution, adding the lysis solution and magnetic beads into a tube strip, adding a sample of virus nucleic acid into the tube strip, and mixing well to lead the magnetic beads to fully absorb the virus nucleic acid; secondly, placing magnetic bead mixed liquor on a magnetic carrier, and allowing the liquor to stand for seconds to separate the magnetic beads and solution; and thirdly, rinsing for protein removal and desalination, and drying in air at room temperature. The method is simple and quick, less apt to cause pollution and high in flexibility. By the method, extracting efficiency is high, nucleic acid purity is high, testing repeatability is good, compatibility to amplification reagents is high, production cost is reduced, operation accuracy is improved, and works such as clinical diagnosis, medicolegal identification and the like are facilitated greatly.

Description

technical field [0001] The invention relates to a method for extracting viral nucleic acid by a one-tube magnetic bead method used for fluorescence quantitative detection, and belongs to the technical field of nucleic acid application in biology. Background technique [0002] Nucleic acid research is an important topic in modern biology and medical research. As the carrier of genetic information, nucleic acid is the material basis of gene expression. In addition to its important role in the growth, development and reproduction of organisms, it is also related to abnormal conditions of life, such as tumor occurrence, radiation damage, and genetic diseases. etc. are also closely related. In medicine, more and more attention has been paid to clinical diagnosis and treatment monitoring, and corresponding methods for the extraction and quantitative detection of viral nucleic acids have also attracted more and more attention. The current method for quantitative detection of vira...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/10C12R1/93
Inventor 朱智芳刘玉方俞萍李晓晨孙克非
Owner TIANGEN BIOTECH BEIJING
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