Supercharge Your Innovation With Domain-Expert AI Agents!

Atypical pneumonia coronary virus gene detecting method and kit

A technology for atypical pneumonia and coronavirus, applied to the determination/testing of microorganisms, biochemical equipment and methods, and resistance to vector-borne diseases, etc., which can solve problems such as false negatives

Inactive Publication Date: 2005-01-26
SHANGHAI FUDAN YUEDA BIOTECH +1
View PDF0 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, this has brought great difficulty to effectively amplifying SARS-specific products, often causing false negative results in experiments

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
  • Atypical pneumonia coronary virus gene detecting method and kit
  • Atypical pneumonia coronary virus gene detecting method and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Concentration of samples:

[0051] Take 170ul of 20% PEG solution and add it to a 1.5ml centrifuge tube, add 500ul of the sample, then add 10.2ul of 20% sulfuric acid, shake and mix for 5s; place overnight at 4°C, centrifuge at 17860g for 20min at 4°C; discard the supernatant , centrifuge at 17860g for 1min at 4°C. Carefully suck off excess supernatant with an adjustable pipette, and pellet for sample extraction.

Embodiment 2

[0053] Samples were extracted using Qiagen Viral RNA Extraction Kit, and operated according to the instructions provided by the manufacturer.

Embodiment 3

[0055] Fluorescent RT-PCR amplification

[0056] Primers used:

[0057] SARS-Fs: 5-ATG, AAT, TAC, CAA, GTC, AAT, GGT, TAC-3 (SEQ ID NO: 1)

[0058] SARS-Fas: 5-CTG, TAG, AAA, ATC, CTA, GCT, GGA, G-3 (SEQ ID NO: 2)

[0059] TaqMan probes use:

[0060] 5'-FAM-TCG, TGC, GTG, GAT, TGG, CTT, TGA, TGT-TAMRA-3' (SEQ ID NO: 5)

[0061] RT-PCR amplification reagents:

[0062] RT-PCR master reaction solution (including primers, dNTP / dUTP, probe, Mg 2+ Wait)

[0063] RT-PCR Enzyme Mix (a solution containing Superscript II reverse transcriptase, Taq enzyme and RNase inhibitor)

[0064] Uracil-N-glycosylase (UNG)

[0065] Operation: Melt 33.8ul RT-PCR main reaction solution, mix well and centrifuge briefly, mix thoroughly with 0.8ul RT-PCR enzyme mixture and 0.4ul UNG, add the processed sample filtrate and working standard to the reaction tube , put the lid on, centrifuge for a few seconds and put it into an iCycler (Roche Company) for amplification.

[0066] Result acquisition: s...

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 sample concentration, RT-PCR amplification, and product detecting method for detecting the SARS coronavirus gene and its kit. The method comprises: (a) concentrating sample by using PEG precipitation method; (b) extracting the virus PNA in the sample, and fluorescent RT-PCR amplification by using specific primer; (c) fluorescent quantitative detection for the amplified product; (d) electrophoresis for the PCR product in step (c). According to the method, the existence of the specific amplified product shows that there is the SARS coronavirus in the sample.

Description

technical field [0001] The invention relates to the technical field of genetic detection, and more specifically to a genetic detection method and a kit for atypical pneumonia coronavirus. Background technique [0002] In 2003, my country and many countries in the world were attacked by a large-scale atypical pneumonia (Atypical pneumonias). Severe acute respiratory syndrome (Severe Acute Respiratory Syndrome, SARS) caused by atypical pneumonia has occurred in 22 countries and regions. The World Health Organization (WHO) quickly established a collaborative research and monitoring network consisting of 13 laboratories in 10 countries around the world after the outbreak. On April 16, WHO officially confirmed that the pathogen of SARS was a new type of coronavirus, which is a single-stranded RNA virus. [0003] There have been more than 8,000 cases of atypical pneumonia in the world, and the death rate in some diseased areas has reached 15%, which has brought great panic to the...

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): C12Q1/68
CPCY02A50/30
Inventor 袁正宏胡芸文
Owner SHANGHAI FUDAN YUEDA BIOTECH
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com