Construction of the Forensic Microbiome Database with Enhanced GeoSourcing

As submitted by the proposer: The forensic community has not yet adopted nor implemented microbiome analysis as a common test in its toolbox. The technology exists and the costs associated with generating next generation sequencing data have decreased significantly; however, the required additional components such as SOPs, validation, basic research, experience, and a database are not in place. We propose laying the framework and design of an open-access microbiome database for the forensic science community – the Forensic Microbiome Database. We will populate the Forensic Microbiome Database (FMD) with hundreds to several thousand 16S datasets and associated metadata availablefrom the public domain. This database will be based on established procedures for database development designed at the J. Craig Venter Institute, incorporating expansive sets of data and metadata that relate to forensics. This objective will be enhanced by writing standards for quality control of the data and the procedures to be used for data entry. In addition, we will sequence and populate the database with 1000 microbiome samples obtained in equal proportions from five (5) distinct regions worldwide. With these data, we will study the ability to perform ‘geosourcing’ by evaluating the sequence signatures from stool and matching oral swab samples and comparing to the origin of sample. By conducting 16S rDNA gene sequencing on these samples and populating the database, we will be able to measure the utility and performance of the database while populating it with very informative data. The scientific accomplishments of this proposal that will benefit the forensic science field include building the first comprehensive forensic microbiome database; providing a host location and continuous monitoring of the database; defining well-structured standard operating procedures (SOPs) for searching against and uploading data into the FMD; testing the utility of the FMD by sequencing a range of samples obtained geographically for querying and proof of concept against the database; and defining SOPs for data generation and inclusion into the FMD. The foundation of this project will serve for future enhancements of the FMD and utility for forensic casework. It will become the standard reference database for interpretation of results and for attributing weight to microbial forensic evidence. It will be developed, put into practice, expanded upon, and advance microbial forensics. This project contains a research and/or development component, as defined in applicable law. ca/ncf