An Automated, All-Ambient Temperature Workflow for the Collection and Storage of Human Blood for DNA and Full Transcriptome Analysis

摘要

Gene expression profiles from blood samples are increasingly used to diagnose and monitor disease progression. However, the utility of blood samples for RNA-based molecular diagnostics depends on a reliable and standardized system for blood collection and preservation during batching and processing. Gene expression profiles in blood specimens are known to be highly labile ex vivo and are highly sensitive to transport and storage conditions, complicating data reliability. In this report, we describe the development and testing of the RNAgard? Blood System, a fully-integrated kit for collection, storage and purification of RNA from human blood under ambient temperature conditions. This system contains a nucleic acid stabilization reagent incorporated into a blood collection tube. The stabilizer immediately preserves RNA and DNA upon blood draw, eliminating dependency on expensive and failure-prone cold-chain logistics. The RNAgard Blood System provides high yields of pure total RNA from human blood stored for up to 14 days at ambient temperature. We describe the optimization of this system for fully automated isolation of highly stable, purified host and pathogen DNA and RNA from a single blood draw tube. The added functionality of high throughput processing in conjunction with co-stabilization of DNA and RNA analytes in the same specimen marks a significant improvement over existing technologies for genetic and molecular analysis.