Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Reagent and method for detecting yellow fever virus

A yellow fever virus and reagent technology, which is applied in the field of temperature amplification technology to detect yellow fever virus, can solve the problems of limited application, specificity, insufficient simplicity of operation, influence of amplification efficiency and speed, etc., and achieves the effect of easy operation.

Inactive Publication Date: 2013-03-06
浙江国际旅行卫生保健中心
View PDF2 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In particular, PCR technology can rapidly amplify trace samples, but since the PCR reaction is divided into three stages of melting, annealing and extension, and only the extension stage performs DNA amplification, the amplification efficiency and speed are greatly affected, and There are deficiencies in specificity and ease of operation, especially the strict requirements on reagents, expensive instruments, and the need for professional personnel to operate, which greatly limits its application in grassroots units and on-site testing.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Reagent and method for detecting yellow fever virus
  • Reagent and method for detecting yellow fever virus
  • Reagent and method for detecting yellow fever virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The detection of embodiment 1 yellow fever virus

[0023] 1. Primer design

[0024] Download representative yellow fever virus sequences from GenBank, select 33 strains of yellow fever virus from 1940 to 2010 and the E gene sequences of vaccine strain yellow fever virus, compare the yellow fever virus E gene sequences with DNAStar software, according to the target Design 4 primers for 6 specific regions in the gene, and design a set of primers (F3, B3, FIP, BIP), which are outer primer B3, outer primer F3, inner primer FIP, and inner primer BIP. The primer gene sequence is as follows:

[0025] Outer primer F3: TGGGAGAGGAGATTCACGT;

[0026] Outer primer B3:TCAAGCCGCCAAATAGCC;

[0027] Inner primer FIP:

[0028] CCACGCCTTTCATGGTCTGAGTTTACCAGTGGCACAAAGAGG;

[0029] Inner primer BIP:

[0030] AGACACCGCCTGGGATTTYAGGCAGAGCCAAAACACCGTATG.

[0031] 2. Extraction of viral RNA

[0032] Dissolve the live attenuated yellow fever virus vaccine with physiological saline acco...

Embodiment 2

[0041] Embodiment 2, detection of yellow fever virus in entry-exit fever personnel

[0042] Use the serum of 23 fever patients who entered the airport port, and use the RNA extraction kit to extract DNA according to the instructions. The RT-LAMP reaction system is as follows: 20mmol / l Tris-HCl, 10mmol / l KCl, 10mmol / l (NH4) 2 SO4, 8mmol / l MgSO 4 , 0.1% Tween-20, the concentration ratio of outer primer and inner primer is 1:8, outer primer F3 and B3 are both 5pmol / l, inner primer BIP and FIP 40pmol / l, 0.8mol / l betaine, dNTP 1.4mmol / l l, 10UAMV reverse transcriptase, 8U Bst-DNA polymerase, 5μl RNA template, complement DEPC-H 2 0 to 25 μl. Sterilized double distilled water was used as negative control, and yellow fever virus vaccine strain was used as positive control. The reaction program was 63°C, 60 minutes. The reaction results of serum samples from 23 patients with fever were centrifuged at 6000rpm, no precipitation was seen for 3 minutes, no green fluorescence was produc...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a reagent for detecting yellow fever virus. The reagent comprises an outer primer F3, an outer primer B3, an inner primer FIP, an inner primer BIP, wherein primer gene sequences are as follows: the outer primer F3: TGGGAGAGGAGATTCACGT; the outer primer B3: TCAAGCCGCCAAATAGCC; the inner primer FIP: CCACGCCTTTCATGGTCTGAGTTTACCAGTGGCACAAAGAGG; and the inner primer BIP: AGACACCGCCTGGGATTTYAGGCAGAGCCAAACACCGTATG. The primers do not have isogeny with nucleotide sequences of other species, so that the specificity of a detection method is ensured. The method for detecting the yellow fever virus through isothermal amplification established by the primers has the advantages of simplicity and convenience in operation, high efficiency, quickness and specificity. Due to the establishment of the method, a blank of the yellow fever virus isothermal nucleic acid amplification detection method is filled, the detection on a sample can be completed by taking around one hour, and significance is provided for preventing the introduction of yellow fever virus in frontier ports.

Description

technical field [0001] The invention relates to a reagent for detecting yellow fever virus and a detection method for yellow fever virus, in particular to a method for detecting yellow fever virus by using isothermal amplification technology. Background technique [0002] Yellow fever virus (Yellow fever virus, YF) is a member of the Flavivirus genus (Flaviridae) in the Flaviridae family. It is the pathogen that causes yellow fever in humans. The disease is mainly prevalent in Africa and South America. It is an acute infection transmitted by mosquitoes. It is one of the three infectious diseases that are subject to international quarantine and monitoring. Humans are universally susceptible to the virus, regardless of age, gender and race. Yellow fever is generally sporadic. If the vector mosquitoes reproduce in large numbers, the infection can cause epidemics among the population. In some outbreaks, the fatality rate can be as high as 20%-40%, which is extremely harmful. T...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/70C12Q1/68G01N21/64
CPCY02A50/30
Inventor 吕沁风郑伟吴忠华曹筝何蕾罗鹏
Owner 浙江国际旅行卫生保健中心
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com