Fluorescent PCR detection method for clostridium difficile toxin genes, as well as primer and kit of fluorescent PCR detection method

A clostridia and detection method technology, applied in the field of genetic detection, can solve problems such as damage, and achieve the effects of avoiding false detection, suitable length, and avoiding lack of specificity

Active Publication Date: 2015-11-11
GUANGZHOU FIRST PEOPLES HOSPITAL (GUANGZHOU DIGESTIVE DISEASE CENT GUANGZHOU FIRST PEOPLES HOSPITAL GUANGZHOU MEDICAL UNIV THE SECOND AFFILIATED HOSPITAL OF SOUTH CHINA UNIV OF TECH) +1
View PDF4 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Exotoxin B is a cytotoxin, and exotoxin B cannot bind directly to mucosal cells in vivo, because exotoxin B can only bind to damaged cells, causing greater damage

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
  • Fluorescent PCR detection method for clostridium difficile toxin genes, as well as primer and kit of fluorescent PCR detection method
  • Fluorescent PCR detection method for clostridium difficile toxin genes, as well as primer and kit of fluorescent PCR detection method
  • Fluorescent PCR detection method for clostridium difficile toxin genes, as well as primer and kit of fluorescent PCR detection method

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach

[0057] 1. Design of specific primers and probes

[0058] 2. According to the nucleotide sequence of Clostridium difficile (C.diff) retrieved from Genbank, design primers and probes for detecting Clostridium difficile toxin A and Clostridium difficile toxin B gene, its sequence is as follows :

[0059] Clostridium difficile toxin A gene

[0060] Upstream primer: tcdA-F: ​​5'-GCAGTCGGATTGCAAGTAATTG-3' (SEQ ID NO.1);

[0061] Downstream primer: tcdA-R: 5'-GTCTGCCAACCTTTTGAGATGAT-3' (SEQ ID NO.2);

[0062] Fluorescent probe: tcdA-P: 5'-VIC-ATTTCAATCCTGACACTGC-MGB-3' (SEQ ID NO.3)

[0063] and

[0064] Clostridium difficile toxin B gene

[0065] Upstream primer: tcdB-F: 5'-TGCAGCCAAAGTTGTTGAATTAGT-3' (SEQ ID NO.4);

[0066] Downstream primer: tcdB-R: 5'-CTGCACCTAAACTTACACCATCTATAATAGT-3' (SEQ ID NO.5);

[0067] Fluorescent probe: tcdB-P: 5'-VIC-TCAACTGCATTAGATGAAA-MGB-3' (SEQ ID NO.6)

[0068] Synthesize the above primers and probes in lifetechnology company;

[0069] 3. 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 belongs to the technical field of gene detection, and specifically relates to a fluorescent PCR detection method for clostridium difficile toxin genes, as well as a primer and a kit of the fluorescent PCR detection method. The primer has good corresponding specificity, proper length and efficiency in transcription, and can not form a secondary structure, the primer contains complementation of not more than four continuous four basic groups; the testing coincidence rate obtained by adopting the primer is 98.6%, and the detection sensitivity can achieve 100copies/mL. According to the detection method, clostridium difficile DNA in a specimen, such as an excrement specimen, can be easily and conveniently extracted, and the genes of clostridium difficile toxins A and B are detected; the specific amplification primer and a fluorescence probe are adopted, so that the detection sensitivity and the specificity are greatly improved, the defect of insufficient specificity existing in an antibody and a cultivation detection method is overcome, and the problem of misdetection is also solved.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to a fluorescent PCR detection method of Clostridium difficile toxin gene, primers thereof, and a kit. Background technique [0002] Clostridium difficile (Clostridium difficile) is an obligate anaerobic bacterium of the genus Clostridium, which is very sensitive to oxygen and difficult to isolate and culture, hence the name. This bacterium was discovered in 1935, but it was not discovered until 1977 that it was related to pseudomembranous colitis caused by long-term clinical use of certain antibiotics (ampicillin, cephalosporin, erythromycin, clindamycin, etc.) Pay attention to. [0003] Clostridium difficile is widely distributed in natural habitats such as soil, hay, sand, feces of some large animals (cows, donkeys, and horses), and in the feces of dogs, cats, rodents, and humans, in addition to being abundant in water and the intestinal tract of animals. Cl...

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): C12Q1/68C12N15/11C12R1/145
CPCC12Q1/6851C12Q1/689C12Q2531/113C12Q2545/113C12Q2563/107
Inventor 聂玉强李永强杜艳雷
Owner GUANGZHOU FIRST PEOPLES HOSPITAL (GUANGZHOU DIGESTIVE DISEASE CENT GUANGZHOU FIRST PEOPLES HOSPITAL GUANGZHOU MEDICAL UNIV THE SECOND AFFILIATED HOSPITAL OF SOUTH CHINA UNIV OF TECH)
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap