Dengue virus gene fragment SERS detection kit and preparation method thereof

A technology for detecting kits and gene fragments, applied in the fields of biological detection and spectroscopy detection, to achieve excellent SERS performance, high sensitivity detection, and improve the detection limit

Active Publication Date: 2019-09-10
NANJING UNIV OF POSTS & TELECOMM
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, both methods are prone to false positive results

Method used

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  • Dengue virus gene fragment SERS detection kit and preparation method thereof
  • Dengue virus gene fragment SERS detection kit and preparation method thereof
  • Dengue virus gene fragment SERS detection kit and preparation method thereof

Examples

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Effect test

Embodiment 1

[0071] Example 1 Preparation and detection method of SERS kit for detection of DENV gene fragments:

[0072] (1) The silver nanorod array SERS substrate is rinsed several times with ultrapure water;

[0073] (2) Add the first reagents L1 and L2 single strands and hairpin structure C1 to the PCR tube, hybridize for 120 min at 25°C;

[0074] (3) Add DENV and the second reagent C2 to the mixture obtained in step (2), and hybridize for 90 minutes at 25° C.;

[0075] (4) Add the third reagent H1 and the fourth reagent H2 to the mixed solution obtained in step (3), and hybridize for 90 minutes at 25° C.;

[0076] The amount of each DNA in the above process ensures that the final concentration of L1, L2, C1, C2, H1 and H2 in the PCR tube is 1 μM.

[0077] (5) Take the mixed solution (1μM) obtained in step (4), drop 20μL into the small hole on the SERS substrate, incubate for 3h at 25°C and 80% humidity, and then wash it with TM buffer (10mM phosphate, 100mM sodium chloride, pH 7.4...

Embodiment 2D

[0079] The preparation and detection method of the SERS kit of embodiment 2DENV gene fragment detection:

[0080] (1) The silver nanorod array SERS substrate is rinsed several times with ultrapure water;

[0081] (2) Add the first reagents L1 and L2 single strands and hairpin structure C1 to the PCR tube, hybridize for 120 min at 25°C;

[0082] (3) Add DENV and the second reagent C2 to the mixture obtained in step (2), and hybridize for 90 minutes at 25° C.;

[0083] (4) Add the third reagent H1 and the fourth reagent H2 to the mixed solution obtained in step (3), and hybridize for 90 minutes at 25° C.;

[0084] The amount of each DNA in the above process ensures that the final concentration of L1, L2, C1, C2, H1 and H2 in the PCR tube is 1 μM.

[0085] (5) Take the mixed solution (1μM) obtained in step (4), drop 20μL into the small hole on the SERS substrate, incubate for 3h at 25°C and 80% humidity, and then wash it with TM buffer (10mM phosphate, 100mM sodium chloride, p...

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Abstract

The invention discloses a dengue virus (DENV) gene fragment SERS detection kit and a preparation method thereof. The invention also discloses a detection method of the dengue virus gene fragment SERSdetection kit. By testing Raman signals of dye molecules ROX on a substrate, the specific and highly sensitive detection of DENV gene fragments is realized. For DENV gene fragment detection, C1, L1, L2, C2, H1 and H2 in a first reagent, a second reagent, a third reagent and a fourth reagent are DNA chains, with different nucleotide sequences, designed for the DENV gene fragments. The detection kitis easy to prepare and high in detection sensitivity and specificity, and has wide application prospects in the fields of dengue virus detection and the like.

Description

technical field [0001] The invention belongs to the fields of biological detection and spectroscopy detection, and relates to a dengue virus gene fragment SERS detection kit and a preparation method thereof. Background technique [0002] Dengue viruses belong to the Flaviviridae family, and four different dengue virus types (dengue 1, 2, 3, 4) can infect humans. These serotypes of dengue viruses can cause mild fever at the time of initial infection and may develop for the more severe forms of dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS), which can lead to death. Over the past 40 years, dengue fever / dengue hemorrhagic fever (DEN / DHF) has spread rapidly worldwide, with increasing frequency and scale of epidemics, and more effective surveillance, prevention and control measures are urgently needed. In most countries where DEN / DHF is endemic, there is no adequate surveillance as a means of assessing disease burden, nor adequate mosquito control or vaccine prev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/682G01N21/65
CPCC12Q1/701C12Q1/682G01N21/65Y02A50/30
Inventor 宋春元汪联辉刘洋张晶晶蒋新宇董晨
Owner NANJING UNIV OF POSTS & TELECOMM
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