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Effective microbiological detection method capable of being applied to oil-gas exploration

A technology for oil and gas and methane oxidizing bacteria, which is applied in the fields of biochemical equipment and methods, and the determination/inspection of microorganisms. effect of risk

Inactive Publication Date: 2012-09-19
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the risk of "false positive" results when methanotrophs are used as indicator bacteria in the current microbial oil and gas exploration, that is, it is impossible to distinguish the source of methane as biological methanogenesis or geographical methanogenesis, the present invention provides a new method that can eliminate biological methanogenesis Abnormal detection method of affected methanotrophs

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] Example 1: The sampling area is a small undeveloped gas field in Shengli Oilfield. The sampling points include the center of the gas field, the boundary of the gas field and two blank controls, which are defined as points A, B, C and D respectively. The sampling depth is 60cm. The distance between each sampling point is about 750m. Soil samples were placed on ice after acquisition, quickly transferred to the laboratory and stored at -20°C.

Embodiment 2

[0011] Example 2: Soil DNA was extracted using FastDNA SPIN Kit for Soil (MP Biomedicals, LLC, USA).

Embodiment 3

[0012] Example 3: Real-time PCR using Mx3005P QPCR Systems (Stratagene). The reagent used is SYBR Premix Ex Taq TM (Fermentas). In the process of using the pmoA gene to quantify methanotrophs, the primers used were A189f (5'-GGNGACTGGGACTTCT GG-3') and mb661R (5'-CCGGMGCAACGTCYTTACC-3'). A standard curve was constructed using uncultured bacteria clone M-5 pmoA gene (GQ906777) as a standard. The program used in Real-time PCR was: 95°C for 10 min, followed by denaturation at 95°C for 20 s, annealing at 57°C for 30 s, and extension at 72°C for 35 s. This cycle was repeated 40 times, and the signal was collected at the end of the annealing stage. When the mcrA gene was used to quantify the methanogens, the primers used were ME3MF (5'-ATGTCNGGTGGHGTMGGSTTYAC-3') and ME2r (5'-TCATBGCRTAGTTDGGRTAGT-3'). A standard curve was constructed using uncultured archaeon clone Ace-28 mcrAgene (EU275997) as a standard. The program used in Real-time PCR was: 95°C for 10 min, followed by de...

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Abstract

The invention discloses an effective microbiological detection method capable of being applied to oil-gas exploration, belongs to the field of biotechnology, and relates to a method for detecting unusual methane-oxidizing bacteria content, which can be applied to microbial oil-gas exploration, takes methane-oxidizing bacteria as indicators and can effectively avoid the influence of biomethanation. The methane-oxidizing bacteria and methanogens in a soil sample are respectively quantified by a Real-time polymerase chain reaction (PCR) technology, the methane-oxidizing bacteria content and ratios of the methane-oxidizing bacteria to the methanogens in different regions are compared, and that whether the reason of unusual methane-oxidizing bacteria is biomethanation is further deduced, so that the influence of the biomethanation is eliminated and risks of false negative results are reduced. The method can be applied to the microbial oil-gas exploration, provides cheap and effective help for the early stage of the exploration, and indicates and predicts favorable exploration blocks to reduce exploration risks and cost.

Description

Technical field: [0001] The patent of the invention belongs to the field of biotechnology, and relates to a method for detecting abnormal content of methane-oxidizing bacteria that can be applied to microbial oil and gas exploration and can effectively avoid the influence of biological methane production. Background technique: [0002] After years of 3D seismic and seismic stratigraphy exploration, a large number of oil and gas reservoir resources that are easy to find have been fully developed, and relatively small, non-structural and subtle oil and gas reservoir resources have become the main targets of exploration. But what followed was a substantial increase in the cost of exploration. Therefore, it is imminent to establish a cheap and effective exploration technology system. In modern exploration methods, microbial oil and gas exploration can provide cheap and effective help for initial exploration, indicate and predict favorable exploration blocks, and reduce explorat...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/64C12Q1/06
Inventor 齐鸿雁满鹏呼庆马安周白志辉庄国强
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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