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A Rapid, Sensitive, Low-Cost Assay for Detecting Hydrogenotrophic Methanogens in Anaerobic Digesters Using
Anna M Alessi1,2, Bing Tao3, Wei Zhang3
1Department of Biology, University of York, Wentworth Way, York YO10 5DD, UK.
Microorganisms
|May 21, 2020
Summary
New Loop-mediated isothermal amplification (LAMP) assays rapidly detect key hydrogenotrophic methanogens in anaerobic digestion (AD). This offers a low-cost, real-time method for optimizing AD processes and gas production.
Area of Science:
- Microbiology
- Biotechnology
- Environmental Science
Background:
- Anaerobic digestion (AD) microbial communities are crucial for methane production, with methanogens playing an essential role.
- Optimizing AD processes often relies on physicochemical parameters, but microbial community analysis is less common due to cost and time constraints.
- Accurate detection of specific microbial genera is needed for effective AD process monitoring and manipulation.
Purpose of the Study:
- To develop and optimize Loop-mediated isothermal amplification (LAMP) assays for the specific detection of four hydrogenotrophic methanogen genera.
- To provide a rapid, low-cost, and sensitive method for assessing the presence/absence of key methanogens in anaerobic digestion systems.
- To explore the potential of LAMP assays for quantitative measurement and real-time monitoring of AD process conditions.
Main Methods:
- Optimization of LAMP assays targeting the archaeal elongation factor 2 (aEF2) gene.
- Detection of four specific hydrogenotrophic methanogen genera: *Methanoculleus, Methanothermobacter, Methanococcus,* and *Methanobrevibacter* spp.
- Assessment of assay sensitivity down to 1 pg of DNA for most tested methanogen species.
Main Results:
- Successfully optimized LAMP assays for the specific detection of the targeted hydrogenotrophic methanogen genera.
- Demonstrated high sensitivity of the LAMP assays, capable of detecting as little as 1 pg of DNA.
- Established the potential for these assays to serve as a presence/absence indicator for crucial methanogens in AD.
Conclusions:
- The developed LAMP assays offer a simple, fast, and affordable method for specific methanogen detection.
- This approach can be a valuable tool for real-time monitoring and optimization of anaerobic digestion systems.
- Further development could enable quantitative measurements, providing deeper insights into AD microbial community function and performance.
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