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Combination gene probe for sulphate reduction pronucleus bioinstrumentation and detecting process thereof

A gene probe and prokaryotic technology, applied in the field of microbial detection, can solve the problems of incomplete detection and time-consuming extinct dilution method, and achieve the effect of low cost and accurate regulation

Inactive Publication Date: 2006-08-23
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the shortcomings of time-consuming and incomplete detection of the widely used extinct dilution method at present, and propose a new combined gene probe and its detection method for the detection of sulfate-reducing prokaryotes in oil field drainage. It can quickly, comprehensively, accurately and conveniently detect SRPs in oilfield drainage, and can realize timely and accurate control of oil production process

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1. The SRPs in the effluent water samples of Gudao Oil Production Plant of Shengli Oilfield are detected:

[0020] 1) Prepare a combined gene probe solution, 8 kinds of probes I-VIII of the present invention are made in relative proportions of 15%, 8%, 16%, 11%, 15%, 10%, 6% and 19% Combined gene probe solution;

[0021] 2) Take a water sample from the oil field drainage, and fix the microbial cells including SRPs in the water sample with a concentration of 2% isopolyaldehyde solution;

[0022] 3) Take 3 μl of the above-mentioned fixed microbial cell water sample and drop it into a glass slide, dry the fixed microbial sample on the glass slide at room temperature, and then use 50%, 80% and 100% alcohol in turn to the glass slide Dehydration of microbial samples on the chip;

[0023] 4) Add 3 μl of the combined gene probe solution to the water sample on the glass slide, add a hybridization solution containing 2.0 M NaCl and 10% formamide, and hybridize at a ...

Embodiment 2

[0027] Example 2: Detection of SRPs in the outflow sample of Shengli Oil Production Plant in Shengli Oilfield

[0028] 1) Prepare a combined gene probe solution, and make a combined gene probe solution with relative ratios of 15%, 30%, 20% and 35% of the four probes I, IV, V and VII of the present invention;

[0029] 2) Take a water sample from the oil field drainage, and fix the microbial cells including SRPs in the water sample with a concentration of 8% isopolyaldehyde solution;

[0030] 3) Take 10 μl of the above-mentioned fixed microbial cell water sample and drop it into a glass slide, dry the fixed microbial sample on the glass slide at room temperature, and then use 50%, 80% and 100% alcohol in turn to the glass slide Dehydration of microbial samples on the chip;

[0031] 4) Add 10 μl of the combined gene probe solution to the water sample on the glass slide, add a hybridization solution containing 5.5M NaCl and 40% formamide, and hybridize at a reaction temperature o...

Embodiment 3

[0035] Example 3: Detection of SRPs in the outflow sample of Dongxin Oil Production Plant of Shengli Oilfield

[0036] 1) Prepare the combined gene probe solution, and make the combined gene probe solution with the relative ratio of 65% and 35% of the two probes IV and VI of the present invention;

[0037] 2) Take a water sample from the oil field drainage, and fix the microbial cells including SRPs in the water sample with a concentration of 10% isopolyaldehyde solution;

[0038] 3) Take 1 μl of the above-mentioned fixed microbial cell water sample and drop it into a glass slide, dry the fixed microbial sample on the glass slide at room temperature, and then use 50%, 80% and 100% alcohol in turn to the glass slide Dehydration of microbial samples on the chip;

[0039]4) Add 1 μl of the combined gene probe solution to the water sample on the glass slide, add a hybridization solution containing 3.5M NaCl and 30% formamide, and hybridize at a reaction temperature of 62° C. for ...

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Abstract

The invention relates to a combined gene probe for sulfate reduction pronucleus biological detection and the detecting method thereof, belongs to microorganism detection technology field. The combined gene probe is composed by at least two gene orders of the 8 ones in SRPs; the detection method comprises: sampling water, fixing the microbial cells in water sample by iso-polyaldehyde solution, instilling it into slide, drying, dewatering, adding the combined gene probe solution, adding hybridization solution of NaCl and formamide, hybriding, washing the probe solution on the slide by phosphate buffer solution, washing the water sample on the slide by distilled water, dying, observing the biological sample on the slide by fluorescence microscope, and counting the SRPs cells with fluorescence. The invention can detect the SRPs in discharged water from oil field rapidly, allsidedly, accurately and conveniently, and can realize the timely and accurate control for oil recovery process.

Description

technical field [0001] The invention belongs to the technical field of microorganism detection, and in particular relates to a method for detecting sulfate-reducing prokaryotes in oilfield drainage by using a combination gene probe using fluorescence in situ hybridization technology of molecular microorganism ecology. Background technique [0002] The detection of sulfate reducing prokaryotes (SRPs) in oilfield drainage is usually limited to the detection of sulfate reducing bacteria (SRB), and the method used is the extinct dilution method (see China Petroleum and Natural Gas Corporation The company's industry standard SY / T5329-94). This method determines the amount of SRB in oil field drainage based on the response of SRB to a specific medium by culturing SRB. Since the various conditions of the natural environment cannot be completely simulated artificially, human beings cannot cultivate all the microorganisms in nature in the laboratory. In fact, only a small amount of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 吴晓磊曾景海屈睿晏芸
Owner TSINGHUA UNIV