| 油气微生物分子勘探技术与初步应用 |
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| 引用本文： | 许科伟,郑旭莹,顾磊,郭嘉琪,汤玉平.油气微生物分子勘探技术与初步应用[J].石油实验地质,2022,44(5):896-903. |
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| 作者姓名： | 许科伟 郑旭莹 顾磊 郭嘉琪 汤玉平 |
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| 作者单位： | 中国石化 石油勘探开发研究院 无锡石油地质研究所, 江苏 无锡 214126 |
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| 基金项目： | 国家自然科学基金项目“页岩气改造—散失途径与保存条件”(41690133)资助。 |
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| 摘 要： | 油气微生物勘探技术因其多解性小、信噪比高、受环境影响小、经济快速的特点，越来越受到勘探家的重视。随着高通量测序、分子生物学技术及生物信息学的快速发展，从微生物群落整体的角度描述特定油气指示微生物的变化，已成为未来微生物勘探技术的重要发展方向。介绍土壤样品采集、DNA提取与分析、判别模型建立和有利区预测等4个油气微生物分子勘探技术关键环节，并针对土壤DNA提取和油气指示微生物检测进行重点阐述。工业级DNA提取技术研究发现，Griffth方法提取并纯化土壤微生物DNA效果最佳。针对大样本量工程，使用高通量移液工作站和商用多孔板DNA抽提试剂盒可大幅提高勘探效率；综合稳定性同位素探针和高通量测序的分子检测技术研究探明了典型含油气区主要油气指示微生物菌种分布规律，高频油指示菌(AMNR族)为节杆菌、分枝杆菌、诺卡氏菌和红螺菌，高频气指示菌为甲基球菌、甲基杆菌、甲基孢囊菌。微生物分子勘探技术在鄂尔多斯盆地杭锦旗地区的初步应用表明，该技术在黄土塬地貌区油气识别精度显著提升，在含油气性评价方面效果良好，为鄂尔多斯盆地北部致密气有利区带筛选提供了参考依据。
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| 关 键 词： | 微生物勘探 分子生物学 高通量测序 稳定性同位素探针 DNA 鄂尔多斯盆地 |
| 收稿时间： | 2022-01-07 |
Molecular microbial prospection for oil and gas and its preliminary application |
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| Affiliation: | Wuxi Research Institute of Petroleum Geology, SINOPEC, Wuxi, Jiangsu 214126, China |
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| Abstract: | Due to the advantages such as low multi-solution, high signal-to-noise ratio, low environmental impact, high efficiency and low cost, microbial prospection for oil and gas has been accepted for petroleum exploreration. With the rapid development of high-throughput sequencing, molecular biology and bioinformatics, describing the changes of specific oil and gas indicator micro-organisms from the perspective of microbial community as a whole has become an valuable progress of microbial exploration method for the future. In this paper, soil sampling, DNA extraction and analysis, discriminant model development and favorable area prediction are introduced, and soil DNA extraction as well as oil/gas indicator micro-organism detection are emphasized. With the study of industrial-grade DNA extraction technology, Griffth method has been considered as the best effect for the extraction and purification of soil microbial DNA. For projects with a large sample number, high-throughput pipetting workstations and commercial multi-well plate DNA extraction kits can greatly improve working efficiency. Integrated stable isotope probes and the molecular detection of high-throughput sequencing showed that the main hydrocarbon indicator microorganism strains typically imply the expected distribution. High frequency oil indicator bacteria (AMNR) are Arthrobacter, Mycobacterium, Nocardia, and Rhodospirillum, while high frequency gas indicator bacteria include Methylococcus, Methylobacterium, and Methylocystis. The preliminary application in the Hangjinqi area shows that the microbial molecular exploration technology can significantly improve the oil and gas identification accuracy in loess tableland regions, and has a good effect on the evaluation of oil and gas bearing property, which provides a reference for the selection of favorable tight gas regions in the northern part of Ordos Basin. |
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