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

Primer and fluorescent probe for detecting sulfate reduction bacterium

A fluorescent probe and sulfate technology, applied in the field of primers and probes for detecting microorganisms, can solve the problems of high non-specificity, low detection results, and long detection time.

Inactive Publication Date: 2007-12-12
HARBIN INST OF TECH
View PDF0 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current quantitative detection methods for SRB bacteria (such as: extinction dilution method, etc.) have the defects of long detection time, high non-specificity, and low detection results

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 and fluorescent probe for detecting sulfate reduction bacterium
  • Primer and fluorescent probe for detecting sulfate reduction bacterium

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach 1

[0009] Specific embodiment one: The upstream primer gene sequence (FP) for detecting sulfate-reducing bacteria in this embodiment is 5'-GTTCCCTGCTCGTGCCCT-3'; the downstream primer gene sequence (RP) is 5'-TTCCTTGAAGAAGATGTACGGGTT-3'.

[0010] In this embodiment, the detection primers are specifically designed and synthesized according to the conserved structural framework of the DSR gene in sulfate-reducing bacteria.

specific Embodiment approach 2

[0011] Specific embodiment 2: The fluorescent probe used to detect sulfate-reducing bacteria in this embodiment consists of a reporter fluorescent group, a quencher fluorescent group and a probe gene sequence; the probe gene sequence is 5'-AATGGTGGATGGAAGAAGGC-3', The reporter fluorescent group labeled at the 5' end of the probe gene sequence is FAM, and the quencher fluorescent group labeled at the 3' end of the probe gene sequence is TAMRA.

[0012] The gene sequence of the fluorescent probe in this embodiment is specifically designed and synthesized according to the conserved structural framework of the DSR gene in sulfate-reducing bacteria.

specific Embodiment approach 3

[0013] Specific embodiment three: this embodiment adopts fluorescent quantitative PCR method to detect sulfate-reducing bacteria:

[0014] 1. Design and synthesis of detection primers and fluorescent probes

[0015] Design and synthesize specific primers and fluorescent probes according to the conserved structural framework of the DSR gene in sulfate-reducing bacteria. The gene sequence of the upstream primer (FP) is 5'-GTTCCCTGCTCGTGCCCT-3'; the gene sequence of the downstream primer (RP) is 5'-TTCCTTGAAGAAGATGTACGGGTT-3'; the gene sequence of the probe is 5'-AATGGTGGATGGAAGAAGGC-3', the gene sequence of the probe is 5' The reporter fluorophore labeled at the 'end is FAM, and the quencher fluorophore labeled at the 3' end of the probe gene sequence is TAMRA. The synthesis of primers and fluorescent probes was done by a bioengineering company.

[0016] 2. Primer and fluorescent probe inspection

[0017] (1) Preparation of standard product (sulfate-reducing bacteria) gene fr...

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 kind of primer and fluorescent probe that are used for sulfate-reducing bacteria detection, relating to primer and fluorescent probe that are used for microbe detection. It overcomes shortcomings of long detection time, high non-specificity and smaller detection result compared to correct result which are existed in current quantitative determination method for SBR bacteria. The gene order of upstream primer is 5'-GTTCCCTGCTCGTGCCCT-3'and gene order of downstream primer is 5'-TTCCTTGAAGAAGATGTACGGGTT-3'. The fluorescent probe comprises report fluorophor, quenching fluorophor and probe gene order; the probe gene order is 5'-AATGGTGGATGGAAGAAGGC-3', the report fluorophor marked by 5'end in probe gene order is FAM, and the quenching fluoropho marked by 3' end in probe gene order is TAMRA. The invention is characterized by high specificity of primer and fluorescent probe, short SBR bacteria determination time, which needs only 6 hours for sample mould DNA extraction to fluorescence quantitative PCR detection.

Description

technical field [0001] The present invention relates to primers and probes for detecting microorganisms. Background technique [0002] Sulfate reducing bacteria (SRB) is a general term for a class of bacteria related to the sulfate reduction reaction. Gram-negative bacteria (such as Desulfovibrio, Desulfobacterium), Gram-positive bacteria (Desulfotomaculum), thermophilic bacteria (Thermodesulfobacterium) and thermophilic archaea ( Archaeoglobus) and other microorganisms. [0003] SRB bacteria reduce sulfate to H through a series of complex biochemical reactions 2 S and lots of sulfides, sulfides and H 2 S not only corrodes oilfield production equipment; its corrosion products (metal sulfides) are insoluble in water, resulting in blackened sewage and increased suspended solids content, causing the suspended solids content in the treated water to exceed the standard, seriously endangering the normal production of the oilfield. Therefore, the rapid and quantitative detectio...

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): C12Q1/68G01N33/52
Inventor 魏利马放李维国
Owner HARBIN INST OF 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