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

Method for detecting arbovirus based on hyper-branched rolling cycle amplification nucleic acid test strip

A technology of nucleic acid test strips and arboviruses, applied in biochemical equipment and methods, resistance to vector-borne diseases, measurement/inspection of microorganisms, etc.

Active Publication Date: 2019-04-16
山东国际旅行卫生保健中心
View PDF7 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the application of this technology in the field of insect-borne pathogen detection is still blank, and has great application prospects

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
  • Method for detecting arbovirus based on hyper-branched rolling cycle amplification nucleic acid test strip
  • Method for detecting arbovirus based on hyper-branched rolling cycle amplification nucleic acid test strip
  • Method for detecting arbovirus based on hyper-branched rolling cycle amplification nucleic acid test strip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] 1. Preparation of DNA sample templates for Japanese encephalitis virus, dengue virus, and chikungunya virus

[0070] 1.1 Design primers for reverse transcription PCR of Japanese encephalitis virus, dengue virus and chikungunya virus

[0071] Japanese encephalitis virus, dengue virus, and chikungunya virus RNA nucleic acid were provided by the Laboratory of International Travel Health Care Center of Shandong Entry-Exit Inspection and Quarantine Bureau; As a template, the amplification primers of the three viruses were designed respectively, and the primer sequences are shown in Table 1:

[0072] Table 1 Primer Sequence

[0073]

[0074]

[0075] 1.2 Preparation of reverse transcription reaction (RT-PCR) system

[0076] The respective genes were amplified by RT-PCR using the onestep RT-PCR kit (TAKARA), and the reaction system was as follows: 1 μL of each upstream / downstream primer (10 μmol / L), 2 μL of the template, 1 μL of PrimeScript 1 Step Enzymemix, 2× 1 step...

Embodiment 2

[0112] The HRCA test strip of the present invention detects the specificity determination of various arboviruses

[0113] Dengue virus (DENV), Japanese encephalitis virus (JEV), Chikungunya virus (Chikungunya fever, CHIKV), West Nile virus (WNV), yellow fever virus ( Yellow fever virus, YFV), promptly use the nucleic acid DNA of five kinds of viruses as template respectively, be used for the specificity verification test of the present invention, wherein dH 2 O is used as a negative control; the DNA template acquisition method and the HRCA reaction system and conditions are as described above, and the results are shown in Figure 14. The three detection probes were spotted on the same nitrocellulose membrane. The preparation method of the HRCA test strips was described above. The amplified products were detected by the pre-prepared HRCA test strips. The results were shown in Figure 18 , illustrate that the method for detecting several arboviruses based on hyperbranched rollin...

Embodiment 3

[0115] Sensitivity determination of HRCA test strip of the present invention for detecting various arboviruses

[0116] Dengue virus (DENV), Japanese encephalitis virus (JEV), and Chikungunya virus (Chikungunya fever, CHIKV) were used respectively. 6 ~10 0 Copies / μL concentration gradient was used to dilute and used as template for HRCA reaction to carry out HRCA amplification reaction; where dH 2 O is used as a negative control; DNA template acquisition method and HRCA reaction system and conditions see before, the results show see Figure 11 , Figure 12, Figure 13 . Spot the three detection probes on the same nitrocellulose membrane. The preparation method of the HRCA detection test strip is described above. The amplified product is detected by the HRCA test strip. The color change of the stain is observed with the naked eye, and Gelpro image analysis can also be used. The software analyzes the speckle value, compares the difference between the speckle value and the nak...

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 discloses a method for detecting Japanese encephalitis virus, Dengue virus and Chikungunya virus based on a hyper-branched rolling cycle amplification nucleic acid test strip, and belongs to the field of entry-exit health inspection and quarantine. The method includes the following steps: (1) designing lock probes and general primers for all the viruses; (2) optimizing the conditionsfor hyper-branched rolling cycle amplification, and establishing a specific and accurate hyper-branched rolling cycle amplification detection technology system for the Japanese encephalitis virus, Dengue virus and Chikungunya virus; and (3) crosslinking the three arboviruses detection probes to a cellulose nitrate membrane to achieve specific detection of viral hyper-branched rolling cycle amplification products, and successfully developing the hyper-branched rolling cycle amplification test strip for detecting various arboviruses. The specificity and sensitivity of detection are ensured, thedetection time is shortened and the custom clearance speed is improved, and the test strip is of great significance for improving interception of entry health epidemics, protecting safety of lives ofChinese people, speeding up the custom clearance speed at ports and reducing the cost of detection.

Description

technical field [0001] The invention relates to the technical field of detection of arbovirus-carried infectious diseases, in particular to a method for detecting a variety of arboviruses based on a hyperbranched rolling circle amplification test strip, which belongs to the category of health and quarantine. Background technique [0002] At present, among the important infectious diseases detected at ports in my country, the viral diseases transmitted by mosquitoes mainly include Japanese encephalitis (also known as Japanese encephalitis), dengue fever, chikungunya fever, West Nile fever, Yellow fever etc. Among them, Japanese encephalitis virus (JEV), dengue virus (DENV), West Nile virus (WNV), and yellow fever virus (YFV) belong to the family Flaviviridae. Chikungunya virus is a member of the alphavirus genus of the Togaviridae family. These five viruses are all RNA viruses, and their genetic structures are very similar. At the same time, these five viruses share vectors....

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844
CPCC12Q1/6844C12Q1/701C12Q2531/125C12Q2521/107Y02A50/30
Inventor 张瑾徐翮飞薛晓宁张娟徐颖
Owner 山东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