一种检测产生硫化氢细菌的灵敏目视方法
June 27th, 2022
在这里，我们提出了一种检测产生硫化氢的细菌的方案，其中包含用于硫化铋（BS）沉淀的改进方案。这种方法的主要优点是易于评估并且不需要专门的设备。
产生硫化氢的细菌可以利用含硫氨基酸和蛋白质来生产硫化氢。在这里，我们建立了一种灵敏而简单的方法来检测细菌中的硫化氢。该方法是硫化铋沉淀的改进版本，使用96孔透明微量滴定板。
将细菌培养物与含有L-半胱氨酸的铋溶液合并培养20分钟，最后观察到黑色沉淀。演示这些程序的将是来自我们实验室的研究助理Wei Zhu。首先将细菌菌落从Luria-Bertani或LB琼脂平板转移到100毫升LB培养基中，并在37摄氏度下培养12至16小时，直到细菌浓度约为每毫升10亿个细胞。
将一个梭杆菌核和普通变形杆菌的细菌菌落从胰蛋白酶大豆肉汤或TSB琼脂平板转移到100毫升TSB培养基中，并在37摄氏度下厌氧培养24小时，直到细菌浓度约为每毫升10亿个细胞。在96孔微量滴定板中将100微升细菌培养物与100微升新制备的铋溶液混合，并在37摄氏度下培养20分钟。对于每种细菌菌株，一式三份进行分析。
20 分钟后，检查颜色是否变化。如果溶液的颜色从浅黄色变为黑色，则表明细菌能够产生硫化氢或H2S。重复此测量三次。
使用不同浓度的硫化氢钠与硫化铋溶液混合确定方法的灵敏度。最后，通过观察黑色硫化铋沉淀物的形成来确定硫化氢铋和硫化物离子的存在。使用从无颜色生产到最深黑色生产的视觉量表对孔的颜色进行评分。
使用选定细菌菌株的纯培养物研究了硫化氢测试的性能。结果表明，副伤寒沙门氏菌B、核梭杆菌、粪肠球菌、铜绿假单胞菌和普通变形杆菌均能产生硫化氢和黑色硫化铋沉淀。而副伤寒沙门氏菌 A、金黄色葡萄球菌、嗜水气单胞菌和肺炎克雷伯菌未显示任何黑色沉淀。
这种方法的主要优点是易于复制并且不需要专门的设备。
View the full transcript and gain access to thousands of scientific videos
概述
本研究提出了一种使用改良的铋硫化物沉淀法检测产硫化氢细菌的敏感且简单的方法。该方法特别有益，因为它不需要专门的设备，并且可以轻松评估。
关键研究组成部分
研究领域
- 微生物学
- 细菌生理学
- 细菌代谢
背景
- 某些细菌在利用含硫氨基酸和蛋白质时会产生硫化氢。
- 检测这些细菌有助于理解它们的代谢途径和潜在的致病性。
- 以往的检测方法可能需要复杂的设置或专门的设备。
使用的方法
- 在96孔透明微孔板中实施改良的铋硫化物沉淀法。
- 使用核糖杆菌和弗氏肠杆菌培养物——这两种都是已知的产硫化氢菌。
- 通过视觉评分颜色变化来识别硫化氢的产生。
主要结果
- 副伤寒沙门氏菌、核糖杆菌、肠球菌、铜绿假单胞菌和弗氏肠杆菌产生了硫化氢，表现为黑色铋硫化物沉淀。
- 其他菌株，包括伤寒沙门氏菌和金黄色葡萄球菌，未能产生沉淀。
- 该方法对产硫化氢细菌显示出高灵敏度和特异性。
结论
- 所介绍的方法提供了一种检测产硫化氢细菌的有效手段。
- 这种技术对于微生物学研究有用，可以应用于各种研究环境。
常见问题
Executive Industry Relevance
Rapid and sensitive detection of hydrogen sulfide-producing bacteria is critical for early-stage target validation and mechanistic de-risking in microbiome and infectious disease research. This high-throughput visual assay enables reliable identification of bacterial metabolic phenotypes, supporting predictive confidence in discovery workflows. Its simplicity and scalability facilitate integration into screening and assay development pipelines across biopharma R&D portfolios.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Enables functional validation of bacterial metabolic pathways relevant to disease models.
- Supports mechanistic de-risking by distinguishing hydrogen sulfide producers from non-producers.
- Facilitates rapid triage of microbial targets for downstream investigation.
Screening & Assay Development
- Provides a standardized, reproducible assay format using 96-well microtiter plates.
- Delivers clear, quantitative visual outputs for high-throughput screening.
- Reduces assay complexity and eliminates the need for specialized instrumentation.
Translational & Preclinical Research
- Aligns microbial phenotype detection with disease-relevant system modeling when hydrogen sulfide is implicated.
- Supports continuity from discovery to preclinical validation by enabling consistent metabolic profiling.
- Improves risk-adjusted advancement decisions for microbiome-targeted programs.
Pipeline & Workflow Integration
This visual detection method fits at the interface of early discovery and screening, enabling rapid hypothesis testing and assay readiness for lead identification and preclinical research.
- Discovery Biology: Supports hypothesis-driven interrogation of bacterial metabolic function.
- Screening: Offers scalable, reproducible outputs for compound or strain evaluation.
- Analytics: Provides quantitative colorimetric readouts for comparative analysis across conditions.
- Translational Research: Facilitates alignment of microbial metabolic activity with disease models when relevant.
- Enterprise Reuse: Establishes a broadly applicable platform for bacterial phenotype screening.
Operational & Enterprise Impact
- Scientific Value: Increases predictive confidence and reduces ambiguity in microbial target validation.
- Operational Value: Enhances standardization, reproducibility, and throughput in assay workflows.
- Strategic Value: Enables more informed go/no-go decisions and efficient resource allocation.
- Portfolio Impact: Supports risk-adjusted prioritization of microbial targets and programs.
Implementation Considerations
- Requires basic microbiological expertise for culture handling and assay setup.
- Utilizes standard laboratory equipment and 96-well plate readers for scalability.
- Demands cross-team agreement on visual scoring criteria for reproducibility.
- Adaptable to various bacterial strains and model systems with minimal protocol modification.
- Visual scoring may require calibration to minimize subjective interpretation.
