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Nucleic acid encapsulating reagent and application thereof

A nucleic acid and reagent technology, applied in the field of nucleic acid encapsulation reagents, can solve the problems of low success rate, time-consuming and labor-intensive, etc., and achieve good adhesion effect, good dissolution effect, and good nucleic acid embedding effect

Active Publication Date: 2017-05-31
CAPITALBIO CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Virus isolation is the gold standard for diagnosing Zika virus infection, but the method is laborious and time-consuming with low success rates

Method used

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  • Nucleic acid encapsulating reagent and application thereof
  • Nucleic acid encapsulating reagent and application thereof
  • Nucleic acid encapsulating reagent and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Embodiment 1, the preparation of packaging reagent

[0061] Encapsulation reagent: composed of 5 parts by mass of agarose, 3 parts by mass of saponin and 4 parts by mass of sodium hydroxypropyl cellulose.

Embodiment 2

[0062] Embodiment 2, preparation for detecting the reaction device of Zika virus

[0063] 1. Mix the encapsulation reagent prepared in Example 1, primer ZIKV-F, primer ZIKV-R, probe ZIKV-P and sterile water to obtain embedding solution. In the embedding solution, the concentration of the encapsulation reagent was 0.1 g / 100 ml, the concentration of the primer ZIKV-F was 2 μM, the concentration of the primer ZIKV-R was 2 μM, and the concentration of the probe ZIKV-P was 1 μM.

[0064] Primer ZIKV-F: 5'-TGCATWGGAGTCAGCAA-3';

[0065] Primer ZIKV-R: 5'-AATTCTAATACGACTCACTATAGGGAGATAGGATCTTACTCVGCCA-3';

[0066] Probe ZIKV-P: 5'-FAM-CGGACGTCAGGTGGGACYTGGGTTGATGTTGTCTTGGGCCTG-BHQ2-3'.

[0067] Primer ZIKV-F is shown as sequence 1 in the sequence listing. The primer ZIKV-R is shown as sequence 2 in the sequence listing. The nucleotide sequence of the probe ZIKV-P is shown as sequence 3 in the sequence listing, the 5' end has a fluorescent group FAM, and the 3' end has a quencher ...

Embodiment 3

[0070] Embodiment 3, the preparation of the kit for detecting Zika virus

[0071] 1. Preparation of kits

[0072] The kit consists of the following components (each component is individually packaged):

[0073] (1) The reaction device for detecting Zika virus prepared in Example 2.

[0074] (2) Amplification reaction solution BQ: the solvent is Tris-HCl buffer solution with pH 8.0 and 100 mM; the solute and its concentration are as follows: 2 mM dNTP (that is, the concentration of dATP, dTTP, dCTP and dGTP is 2 mM), 4 mM rNTP ( That is, the concentrations of rATP, rTTP, rCTP and rGTP are all 4mM), 50mM DTT, 60mM MgCl 2 , 300mM KCl, 15% (volume percentage content) DMSO, 1M sorbitol.

[0075] (3) Enzyme mixture EQ: the solvent is water; the solute and its concentration are as follows, AMV reverse transcriptase 0.8U / μl, T7 RNA polymerase 3.5U / μl, ribonuclease H 0.3U / μl, pyrophosphatase 0.5U / μl μl, RNase inhibitor 5U / μl, BSA 0.5μg / μl.

[0076] (4) Positive quality control pro...

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Abstract

The invention discloses a nucleic acid encapsulating reagent and an application thereof. The nucleic acid encapsulating reagent provided by the invention consists of 3-10 parts by mass of agarose, 1-5 parts by mass of saponin and 2-6 parts by mass of hydroxy propyl cellulose. The invention also discloses a target material detection device, wherein the target material detection device is obtained by embedding primers and a probe for detecting a target material in a reaction device by virtue of the nucleic acid encapsulating reagent. The invention also discloses a Zika virus detection device, wherein the Zika virus detection device is obtained by embedding a primer ZIKV-F as shown in a sequence 1, a primer ZIKV-R as shown in a sequence 2 and a probe ZIKV-P as shown in a sequence 3 in the reaction device by virtue of the nucleic acid encapsulating reagent prescribed in any one of previous claims. The invention has an important application value for detecting the target material with the application of a nucleic acid sequence-based amplification technology. The invention has great application and popularization value for detection of Zika viruses and for prevention and control of related diseases.

Description

technical field [0001] The invention relates to a nucleic acid encapsulation reagent and its application. Background technique [0002] Zika virus (ZIKV) is a virus transmitted by the bite of an Aedes mosquito. In 2015, Brazil discovered the first case of Zika virus infection, and then the epidemic continued to ferment until a large-scale Zika virus outbreak broke out, which quickly spread to more than 30 South American and African regions. So far, more than 1.5 million people have been infected with Zika virus. On February 2, 2016, the World Health Organization held an emergency meeting and decided to list the spread of Zika virus as a global public health emergency. The Chinese Health and Family Planning Commission announced the "Diagnosis and Treatment Plan for Zika Virus Disease" (1st Edition, 2016) on February 3, 2016. Six days later, my country confirmed the first case of imported Zika virus infection. So far, my country has confirmed Multiple imported cases. At the ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12M1/00C12Q1/70C12Q1/68C12R1/93
CPCC12N15/11C12Q1/701
Inventor 张岩盖伟邢婉丽宋翠丹程京
Owner CAPITALBIO CORP
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