Method for extracting specific nucleic acid in blood screening reagent

An extraction method and specific technology, applied in chemical instruments and methods, biochemical equipment and methods, preparation of sugar derivatives, etc., can solve the problems of false positive test results, processing interference, etc., to improve specificity and avoid false positives , the effect of accurate test results

Inactive Publication Date: 2009-08-05
HANGZHOU BIOER TECH CO LTD
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  • Abstract
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Problems solved by technology

[0004] Since the surface of ordinary micro-nano magnetic beads is coated with a layer of silicate material, it binds to the nucleic acid in the sample through hydrogen bonding, van der Waals force, etc., and can absorb all nucleic acids during the extraction and purification process, which is a non-specific extraction. After elution, a mixture of various nucleic acids is obtained, which will cause a lot of interference to subsequent PCR amplification and processing of other samples, especially in blood screening reagents that require high sensitivity, due to the different nucleic acid sequences of various viruses Homology causes false positive test results

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  • Method for extracting specific nucleic acid in blood screening reagent
  • Method for extracting specific nucleic acid in blood screening reagent
  • Method for extracting specific nucleic acid in blood screening reagent

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[0019] The present invention is further described below in conjunction with the examples.

[0020] 1) Purchase 1.5-100 μm micro-nano magnetic beads containing streptavidin, and synthesize the specific nucleic acid sequences of HBV, HCV and HIV as follows:

[0021] HBV-specific nucleic acid sequence: 5'-TGCTGCTGCCTCATCCTTGTC[biotin]-3'

[0022] HCV specific nucleic acid sequence: 5'-AGTAGTTCCTCACAGGGGAGT[biotin]-3'

[0023] HIV-specific nucleic acid sequence: 5'-TGGCTGCTTGATGTCCCCCCAC[biotin]-3'

[0024] Coat HBV, HCV and HIV-specific nucleic acid sequences on micro-nano magnetic beads according to the instructions provided by commercial magnetic beads;

[0025] 2) Add 0.9ml of mixed blood sample and 0.9ml of LH lysis solution to a tube with a capacity of more than 2ml with a pipette, mix well with a shaker, heat at 75°C for 12min, and then cool at room temperature for 10min.

[0026] Add 50 μl of the magnetic beads of step 1 to the lysed blood sample, and mix at room temper...

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Abstract

The invention discloses an extraction method for specific nucleic acid in a blood screening reagent. The method comprises the following steps: 1.5 to 100 microns of micro-nano-magnetic beads containing chain avidin are coated with HBV, HCV and HIV specific nucleic acid sequences; the micro-nano-magnetic beads coated with the specific nucleic acid sequences are hybridized with the nucleic acid from the disrupted HBV, HCV and HIV viruses in blood samples; a nucleic acid purification instrument is used to separate the magnetic beads from the blood samples, the magnetic beads are washed with so that elute HBV, HCV and the HIV virus nucleic acids are washed off the magnetic beads; and the content of viruses is detected by fluorescence quantitative PCR. The adopted method is favorable for improving the specificity of nucleic acid extraction, avoids the false-positive detection results caused by the homology of all types of virus nucleic acid sequences so that to more accurate detection results are obtained, and can be widely used in blood screening.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the design of specific nucleic acid sequences, the coating method of nano-magnetic beads, and the lysis of viruses. Background technique [0002] The nucleic acid purification technology based on magnetic particles (micro-nano magnetic beads) uses hydrophilic magnetic particles to adsorb nucleic acid molecules in the sample solution, and separates the nucleic acid-loaded magnetic particles from the sample solution under the action of an external magnetic field. , adding an appropriate solution to desorb the nucleic acid from the magnetic particles to obtain purified nucleic acid. [0003] Magnetic particles (micro-nano magnetic beads) made of γFe 2 O 3 and Fe 3 O 4 Homogeneous, superparamagnetic, single-divergent polymeric microspheres synthesized from magnetic materials. Each microsphere is coated with a layer of polymeric material that acts as a carrier for adsorpti...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C07H21/00C07H1/06
Inventor 杨君厉永纲
Owner HANGZHOU BIOER TECH CO LTD
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