The use of molecular biological tools (MBTs) for the detection and quantification of microbial biomarkers (e.g., DNA/RNA) in environmental samples is rapidly increasing as remedial practitioners seek to improve the design, field performance, and monitoring of bioremediation.
This project focused on identifying and minimizing the causes of variability during quantitative real-time polymerase chain reaction (qPCR) enumeration of genes of interest in groundwater, with the goal of developing a foundation for the development of standardized methods for collection, preservation, transport, storage, and processing of environmental qPCR samples collected from contaminated sites.
A technology review on the status of MBTs was performed at the beginning of the project to determine MBT use in other industries. The review focused project goals and activities which included:
Significant results of the project included:
MBTs such as qPCR have the potential to accurately enumerate microbial cells, including Dhc, in groundwater samples if proper procedures and appropriate controls are integrated into the process. The development of optimized methods, including the use of a project developed MIAC, allowed significant improvements in data quality and the ability to better detect and correct for matrix interference. Understanding the distribution of Dhc between sediment and groundwater will allow more accurate conclusions to be drawn regarding total Dhc biomass in aquifers using groundwater data. The ability to better quantify Dhc at a variety of sites with high precision, in multiple labs, will increase remediation practitioners’ confidence in remediation focused qPCR analysis and their ability to manage bioremediation projects effectively.