An automatic detection method for the abundance of butane-oxidizing bacteria

Abstract

The invention discloses a method for automatically detecting the abundance of butane oxidizing bacteria. The method comprises the steps: constructing an automatic workstation of an original-state microbial oil gas and hydrate prospecting technique, performing sample processing, completing liquid transfer by adopting an automatic liquid processing device of the workstation, extracting and purifying DNA (deoxyribonucleic acid) by using an automatic nucleic acid extraction device, performing electrophoresis detection with agarose, performing fluorescent quantitive PCR (polymerase chain reaction) detection of the abundance of butane oxidizing bacteria, obtaining results, and the like. Based on an automatic analysis method, automation of the nucleic acid extraction and purification steps of butane oxidizing bacteria is achieved; automation of the preparation of a specific gene PCR reaction system is achieved, errors caused by human factors introduced by manual operation are reduced, the operation steps are carried out in a clean environment, the whole detection process is controlled by computer software, the detection procedure can be traced, and uniform correction of system errors can be realized.

Description

technical field [0001] The invention relates to the technical field of oil gas and hydrate exploration and oil gas and hydrate reservoir characterization, and more specifically relates to an automatic detection method for the abundance of butane-oxidizing bacteria. Background technique [0002] China is a big energy-consuming country. In 2013, my country's dependence on foreign oil was close to 60%. With the acceleration of industrialization and urbanization, my country's demand for oil, gas and hydrate energy has entered a period of continuous growth. Although my country is implementing strategies for energy conservation, emission reduction and new energy development, my country still cannot get rid of its dependence on oil and gas resources for a long period of time. Therefore, my country's energy security has been put on the important agenda. In order to alleviate my country's energy tension, it is imminent to intensify exploration and develop oil, gas and hydrate more ...

AI Technical Summary

Problems solved by technology

However, because more than 99% of the microorganisms in soil, sediment and water cannot be cultivated, the traditional microbial exploration method based on the culture method is difficult to meet the exploration requirements in terms of detection accuracy; because gaseous hydrocarbon micro-leakage is dynamic and discontinuous , so traditional microbial exploration methods cannot find micro-seepage of gaseous hydrocarbons in blocks where micro-cracks are in a squeezed state; it is difficult to carry out exploration in today's deserts, Gobi and deep ocean environments with a single anomalous index of live bacteria; Two samplings are required for oil and gas reservoir characterization, which not only significantly increases the cost, but also cannot be implemented in many production areas, because most oil and gas reservoirs in my country have not carried out microbial characterization

[0004] The immaturity of microbial oil and gas and hydrate exploration technology is mainly manifested in: (1) In the face of complex samples of obligate or facultative microorganisms with low abundance and indicative significance for oil, gas and hydrate, based on microbial culture methods, The established traditional microbial oil and gas and hydrate exploration technology is not only difficult to measure the microbial abundance (because more than 99% of the microorganisms in the soil and sediments cannot be cultivated), but the data reliability is poor; (3) In the face of complex and diverse oil, gas and hydrate exploration areas, especially deserts, Gobi, saline-alkali lands and ocean deep water areas, traditional microbial oil, gas and hydrate reservoirs established based on microbial culture methods Exploration technology is basically ineffective; the main reason is that the traditional microbial oil, gas and hydrate exploration technology only relies on the abnormality of live bacteria with a single index; (4) The traditional oil, gas and hydrate exploration technology based on culture method has very low sample processing efficiency , it usually takes at least 14 days for a batch of samples to obtain the results, which has long cycle, low efficiency and high cost; (5) There are many factors affecting the data and large fluctuations, which will introduce a large number of human errors in the analysis and determination process; the detection process is not traceable, And it is impossible to correct the system error

[0005] Based on the original state detection technology of microbial single cells and genes, Wang Jianghai and others took the lead in proposing the concept, principle and technical scheme of original state microbial oil and gas and hydrate exploration technology, and performed specific fluorescent staining, methane- Significant progress has been made in the rapid quantitative detection of dead and live bacteria of butane oxidizing bacteria, the establishment of multiple indicators for microbial oil, gas and hydrate exploration, and the PCR strategy for denaturing gradient gel electrophoresis analysis of microbial communities in oil, gas and hydrate fields, forming the following Gaseous hydrocarbon oxidizing bacteria and their associated bacteria, live bacteria, dead bacteria and total bacteria abnormalities are multiple exploration indicators for microbial oil and gas and hydrate exploration technology, but manual methods are still required to detect microbial abundance, which cannot fundamentally solve the problem of microbial oil and gas. Disadvantages of hydrate exploration techniques

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