Primer and fluorescent probe for detecting sulfate reduction bacterial

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: 2007-12-19
HARBIN INST OF TECH
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  • 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

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

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Experimental program
Comparison scheme
Effect test

specific Embodiment approach 1

[0009] Specific embodiment 1: The upstream primer gene sequence (FP) for detecting sulfate-reducing bacteria in this embodiment is 5'-CGCTGAAATGACCATGATGG-3'; the downstream primer gene sequence (RP) is 5'-CGCGCATTTCACGAAGC-3'.

[0010] In this embodiment, the detection primers are specifically designed and synthesized according to the conserved structural framework of the APS 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'-CTGAAGCCTTACGAAGATCG-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 APS 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 APS gene in sulfate-reducing bacteria. The gene sequence of the upstream primer (FP) is 5'-CGCTGAAATGACCATGATGG-3'; the gene sequence of the downstream primer (RP) is 5'-CGCGCATTTCACGAAGC-3'; the gene sequence of the probe is 5'-CTGAAGCCTTACGAAGATCG-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 fragments. ...

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Abstract

The upstream pimer for testing sulfate reducing bacterium is 5-CGCTGAAATGACCATGATGG-3, and its downstream primer is 5-CGCGCATTTCACGAAGC-3. The fluorescent probe is composed of reporter fluorescent group, quenching fluorescent group and probe gene sequence, which being of 5-CTGAAGCCTTACGAAGATCG-3. Said 5 end of probe gene sequence labels that reporter fluorescent group is FAM; and said 3 end of probe gene sequence labels that quenching fluorescent group is TAMRA. This invented primers and fluorescent probe have advantages: high specificity; short-term testing of sulfate bacterium for fluorescent quantitative PCR test, and needing only 6 Hrs. after DNA extraction and fluorescent quantitative PCR test.

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

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/52
Inventor 魏利马放李维国
Owner HARBIN INST OF TECH
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