Kit and method for extracting viral nucleic acid

A virus nucleic acid and kit technology, which is applied in biochemical equipment and methods, microbial measurement/inspection, DNA preparation, etc., can solve the problems of cumbersome operation, pollution, and long time consumption, and achieve high nucleic acid recovery rate and operation flow Clean, time-saving effect

Inactive Publication Date: 2018-10-12
GUANGZHOU YIXIN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The traditional nucleic acid extraction method is cumbersome and time-consuming. For example, the silica gel membrane adsorption method requires multiple centrifugations, which is prone to problems such as cross-contamination
However, the current magnetic bead virus nucleic acid extraction kits on the market are mainly for small-scale and small-volume tests. For blood bank or clinical batches of physical examination samples, it is impossible to quickly extract them at one time, which takes a long time and requires many points. extraction, the risk of contamination increases

Method used

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  • Kit and method for extracting viral nucleic acid
  • Kit and method for extracting viral nucleic acid
  • Kit and method for extracting viral nucleic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A kind of embodiment of the kit for extracting viral nucleic acid in blood screening mixed test sample of the present invention, the composition that comprises and concentration thereof are as follows:

[0037]

[0038]

[0039] Eluent: 10mM Tris-HCl (pH8.0)-DEPC aqueous solution;

[0040] Magnetic beads: commercial silica magnetic beads;

[0041] Proteinase K: commercial product, dissolved in DEPC water, the final concentration is 20mg / mL;

[0042] Nucleic acid precipitation aid: commercial Acryl Carrier.

Embodiment 2

[0044] An embodiment of the method for extracting viral nucleic acid in the blood screening mixed sample of the present invention comprises the following steps:

[0045] 1) Sample preparation: inactivated serum or plasma from blood collection stations or clinical samples, 200 μL each, 12 samples combined into 1 2.4 mL sample to be tested, marked and numbered respectively, and samples less than 2.4 mL were identified Negative blank plasma complement.

[0046] 2) Positive control and negative control samples are set; among them, the positive control sample is the inactivated plasma that has been confirmed as positive for hepatitis B virus by the blood station, and the negative control sample is the inactivated plasma that has been confirmed as negative for hepatitis B virus by the blood station;

[0047] 3) Instrument operation stage:

[0048] (1) Add 3mL rinse solution 1 to the deep well plate 1, and place it on the corresponding slot of the instrument;

[0049] (2) Add 3mL r...

Embodiment 3

[0061] A kind of embodiment of the method for rapid detection of virus in blood plasma / serum of the present invention, comprises the following steps:

[0062] 1. Reagent preparation: Prepare the solution required for the experiment according to Example 1, including lysis solution, binding solution, rinse solution 1, rinse solution 2, eluent, proteinase K solution;

[0063] 2. Sample preparation: take the hepatitis B quality control substance and dilute it with negative plasma to 10 3 IU / mL, dilute 50mL for use;

[0064] 3. Viral DNA extraction:

[0065] (1) Add 3 mL of rinse solution 1 to the deep well plate 1, and put it on the corresponding slot of the instrument (same as Example 2);

[0066] (2) Add 3mL rinse solution 2 to the deep well plate 2, and place it on the corresponding slot of the instrument;

[0067] (3) Add 200 μL of eluent to the elution tube and place it on the corresponding card slot of the instrument;

[0068] (4) Put the magnetic cover plate on the corr...

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Abstract

The invention discloses a kit and method for extracting viral nucleic acid from a blood screening mixed detection sample. The kit comprises a lysate, a binding solution, a first rinsing solution, a second rinsing solution, an eluent, silicon dioxide magnetic beads, proteinase K and nucleic acid settling agent, wherein the first rinsing solution contains the binding solution and sodium citrate. Theinvention further discloses a method for rapidly detecting viruses in plasma / serum. With the kit and the method provided by the invention, trace of viral nucleic acid can be easily captured from thesample under the specific pH, and the recovery rate of the obtained nucleic acid is high; the operation procedure is concise, the extraction and purification of viral nucleic acid can be completed within 30min, and thus the kit is more time-saving and labor-saving than the existing majority of commercial kits; the aimed sample volume dose is large, combined extraction can be carried out on small-volume samples, thus time is saved, and the reagent is saved.

Description

technical field [0001] The invention relates to the technical field of nucleic acid purification, in particular to a method and a kit for extracting viral nucleic acid in serum / plasma. Background technique [0002] The extraction and purification of nucleic acid is the basis for various researches in molecular biology, especially in the field of molecular diagnosis, PCR, RT-PCR, Northern blot, RT-qPCR, cDNA library construction, enzyme digestion and other technical means require high purity, complete good nucleic acid. However, due to the existence of exogenous and endogenous nucleases and their relatively stable enzyme activity, it is relatively difficult to extract and purify high-quality nucleic acids. The formula of nucleic acid extraction reagents has a great influence on the quality and yield of nucleic acid. Therefore, the research and development of nucleic acid extraction reagents with high efficiency, rapid recovery, wide range of recovery and high yield is a subj...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12Q1/6806C12Q1/6851C12Q1/70
CPCC12N15/1013C12Q1/6806C12Q1/6851C12Q1/701C12Q2531/113C12Q2563/107
Inventor 陈永恒谭杰峰宋卉周秋霞
Owner GUANGZHOU YIXIN BIOTECH CO LTD
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