Primer for detecting sulfate reduction bacterium

A sulfate and gene sequence technology, applied in the direction of biological testing, material inspection products, etc., can solve the problems of high non-specificity, low detection results, long detection time, etc.

Inactive Publication Date: 2010-05-26
HARBIN INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 for detecting sulfate reduction bacterium
  • Primer for detecting sulfate reduction bacterium
  • Primer for detecting sulfate reduction bacterium

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Experimental program
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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...

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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 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 detection of sulfat...

Claims

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Application Information

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