title: Bacterial RNA extraction and purification from whole human blood using isotachophoresis.
abstract: We demonstrate a novel assay for physicochemical extraction and isotachophoresis-based purification of 16S rRNA from whole human blood infected with  Pseudomonas putida . This on-chip assay is unique in that the extraction can be automated using isotachophoresis in a simple device with no moving parts, it protects RNA from degradation when isolating from ribonuclease-rich matrices (such as blood), and produces a purified total nucleic acid sample that is compatible with enzymatic amplification assays. We show that the purified RNA is compatible with reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and demonstrate a clinically relevant sensitivity of 0.03 bacteria per nanoliter using RT-qPCR.
authors: Department of Mechanical Engineering, Stanford University, Stanford, California 94305, USA.
authors: Anita
authors: A
authors: Rogacs
authors: Yatian
authors: Y
authors: Qu
authors: Juan G
authors: JG
authors: Santiago
mesh_terms: D006801:Humans; D057625:Isotachophoresis / Q000379:methods*; D016133:Polymerase Chain Reaction; D016958:Pseudomonas putida; D012329:RNA, Bacterial / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism
chemical_list: D012329:RNA, Bacterial; D012260:Ribonucleases