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

Primer for detecting Escherichia coli, and method and application of primer

A technology for Escherichia coli and detection results, which is applied to the detection of Escherichia coli primers and its application fields, can solve the problems of time-consuming, labor-intensive, poor specificity, and inability to achieve rapid goals, and achieve accurate detection results

Inactive Publication Date: 2015-08-26
KUNMING UNIV OF SCI & TECH
View PDF5 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Traditional detection methods for Escherichia coli include conventional smear microscopy and culture methods, but these methods have certain defects. For example, conventional smear microscopy is the most basic bacteriological examination method, but it has low sensitivity and poor specificity; The culture method is currently the main way and means to find the source of infection clinically, but it usually takes 4 to 7 days, the operation is cumbersome, time-consuming and labor-intensive, and cannot achieve the rapid goal

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
  • Primer for detecting Escherichia coli, and method and application of primer
  • Primer for detecting Escherichia coli, and method and application of primer
  • Primer for detecting Escherichia coli, and method and application of primer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Specific target gene screening and primer design:

[0025] 1) Local BLAST analysis

[0026] From the whole genome sequence of Escherichia coli downloaded from NCBI (http: / / www.ncbi.nlm.nih.gov / genome / ), perform a local BLAST search on the local nucleic acid database, and obtain the sequence fragments of Escherichia coli for comparison with the database result.

[0027] 2) BLAST reconfirmation

[0028] Using the online BLAST function, 2 strategies were used to confirm their species specificity and E. coli identification availability. One strategy is to exclude Escherichia coli during BLAST, and the results show that those without any similar sequences are specific genes of Escherichia coli; the second strategy is to search within Escherichia coli during BLAST, and the results return a large number of similar sequences shared by different Escherichia coli strains sequence. Combining the two strategies, the finally obtained gene LafF as shown in SEQ ID NO: 3 ...

Embodiment 2

[0031] The establishment of embodiment 2 detection method

[0032] 1) Extraction of DNA

[0033] Use Beijing Zhuangmeng International Biogene Technology Co., Ltd. Bacterial Genomic DNA Small Extraction Kit for extraction and recovery, the steps are as follows:

[0034] (1) Take 5mL of bacterial culture solution, centrifuge at 12,000rpm for 1 minute, and suck up the supernatant as much as possible.

[0035] (2) Add 500 μL of cell suspension to the centrifuge tube with the bacterial pellet left, use a pipette or a vortex shaker to thoroughly suspend the bacterial cell pellet, and incubate at 37°C for 30 minutes. Mix by inversion several times every 10 minutes. Centrifuge at 12,000rpm for 2 minutes, and try to suck up the supernatant.

[0036] (3) Add 225 μL of buffer A to the cell pellet and shake until the cell is completely suspended.

[0037] (4) Add 6 μL of RNaseA solution to the tube, shake for 15 seconds, and place at room temperature for 5 minutes.

[0038] (5) Add 1...

Embodiment 3

[0059] Embodiment 3 specific detection

[0060] 1. Collect 3 E. coli clinical isolates and 9 non-E. coli strains. The genomes of these strains were extracted with the Bacterial Genomic DNA Small Extraction Kit of Beijing Zhuangmeng International Biogene Technology Co., Ltd. The steps are as follows:

[0061] (1) Take 5mL of bacterial culture solution, centrifuge at 12,000rpm for 1 minute, and suck up the supernatant as much as possible.

[0062] (2) Add 500 μL of cell suspension to the centrifuge tube with the bacterial pellet left, use a pipette or a vortex shaker to thoroughly suspend the bacterial cell pellet, and incubate at 37°C for 30 minutes. Mix by inversion several times every 10 minutes. Centrifuge at 12,000rpm for 2 minutes, and try to suck up the supernatant.

[0063] (3) Add 225 μL of buffer A to the cell pellet and shake until the cell is completely suspended.

[0064] (4) Add 6 μL of RNaseA solution to the tube, shake for 15 seconds, and place at room temper...

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 primer for detecting Escherichia coli, and a method and an application of the primer. The nucleotide sequence of the primer is indicated in SEQ ID NO: 1 and SEQ ID NO: 2, an Escherichia coli detection method is established for the involved primer, and the detection method has the advantages of fine specificity, good sensitivity, rapidness and reliability in detection and simplicity in operation. A kit comprising the primer is designed, and the Escherichia coli is conveniently detected. The specific primer is designed for specific genes LafF of the Escherichia coli and can be used for rapidly, simply, conveniently and accurately detecting the Escherichia coli.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a primer for detecting Escherichia coli and its application. Background technique [0002] Escherichia coli (Escherichia coli) is customarily called Escherichia coli, which belongs to the family Enterobacteriaceae and belongs to the genus Escherichia. Most Escherichia coli are normal resident bacteria in the intestines of humans and various animals, and the detection of Escherichia coli in food or water means direct or indirect recent fecal contamination. Escherichia coli is used as a hygienic bacteriological indicator of fecal pollution in drinking water and food, and its survival time in the outside world is similar to that of some major enteric pathogens. Its appearance may also indicate the presence of certain intestinal pathogens (such as Salmonella, Shigella). Escherichia coli is an internationally recognized indicator bacteria for health monitoring, so the detection technology...

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): C12Q1/68C12Q1/10C12N15/11
CPCC12Q1/686C12Q1/689
Inventor 毛小琴刘有福宋玉竹牛华夏雪山
Owner KUNMING UNIV OF SCI & TECH
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