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

Method for detecting Chikungunya virus nucleic acid by real-time fluorescent quantitative PCR

A technology for real-time fluorescence quantification and viral nucleic acid, applied in the fields of fluorescence/phosphorescence, microbial measurement/inspection, biochemical equipment and methods, etc.

Inactive Publication Date: 2011-01-05
CHINESE ACAD OF INSPECTION & QUARANTINE
View PDF7 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because PCR amplification technology often brings specificity problems, scientists and technicians from all over the world are trying to improve the specificity and sensitivity of PCR technology based on PCR technology and other new technologies.

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 Chikungunya virus nucleic acid by real-time fluorescent quantitative PCR
  • Method for detecting Chikungunya virus nucleic acid by real-time fluorescent quantitative PCR
  • Method for detecting Chikungunya virus nucleic acid by real-time fluorescent quantitative PCR

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Detection of Primer Sensitivity

[0031] 1) Select a highly conserved region fragment in the whole genome of the virus as a standard, and its viral RNA from 2.3 × 10 10 copy / uL was sequentially diluted to 2.3×10 -1 copy / uL, select 2.3×10 6 copy / uL to 2.3×10 -1 The copy / uL copy number gradient is used for detection, the experiment is repeated 3 times, and the average value is taken;

[0032] 2) Negative control: nuclease-free water;

[0033] 3) The reaction system is prepared according to Table 2;

[0034] 4) At the same time, it is compared with the currently commercially available chikungunya virus nucleic acid detection kit.

[0035] Test results such as figure 2 , image 3 , Table 3, Table 4 shown.

[0036] figure 2 Among them, the curves 1-7 are respectively the respective dilutions of the standard substance in the present invention 2.3×10 6 copy / uL-2.3×10 0 The curve of copy / uL amplification.

[0037] image 3 Among them, the curves 1-3 r...

Embodiment 3

[0044] Embodiment 3: Primer detection unknown sample

[0045] 1) Detect the collected unknown samples, screen the Chikungunya virus and monitor the non-specificity of the primers;

[0046] 2) Positive control: virus standard (2.3×10 3 copy / uL);

[0047] 3) The reaction system was prepared according to Table 2.

[0048] See the test results Figure 4 and Table 5.

[0049] Figure 4Only one amplification curve appeared in the positive control, and the rest of the samples had no amplification. The reaction was negative. PTC was used as a positive control.

[0050] The data in Table 5 shows that the results of testing unknown samples were all negative.

[0051] Table 6

[0052]

[0053] Result analysis:

[0054] After the amplification reaction completed 40 cycles, the minimum detection limit of the primer and probe detection positive standard in the present invention was 2.3copy / uL, while the minimum detection limit of the currently commercially available chikungunya...

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 Chikungunya virus nucleic acid by real-time fluorescent quantitative PCR, comprising: obtaining a primer and a probe, and detecting the Chikungunya virus nucleic acid. The invention has the advantages of good specificity, high sensitivity and the like.

Description

technical field [0001] The invention provides a method for detecting chikungunya virus nucleic acid by real-time fluorescent quantitative PCR. Background technique [0002] Chikungunya virus (Chikungunya virus) is a single-stranded positive-sense RNA virus belonging to the Togaviridae family (Togaviridae) Alphavirus (Alphavirus), causing Chikungunya fever (Chikungunyafever, CHIK). The disease is an acute infectious disease, transmitted to humans by the Aedes mosquito, clinically characterized by fever, joint pain, rash, and mild bleeding. [0003] Chikungunya virus was first isolated from the blood of a febrile patient in Tanzania in 1953. From the 1960s to the 1990s, the disease was prevalent in many countries in central and southern Africa, and CHIK virus was isolated, such as Sudan, Uganda, Democratic Republic of the Congo, Central African Republic, Malawi, Zimbabwe, Kenya and South Africa; Senegal, Benin, the Republic of Guinea, Côte d'Ivoire, and Nigeria in West Afric...

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): C12Q1/70C12Q1/68G01N21/64
CPCY02A50/30
Inventor 姚李四燕清丽张晓龙
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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