title: Amplification of viral RNA from drinking water using TransPlex™ whole-transcriptome amplification.
abstract: OBJECTIVE
  Viral pathogens in environmental media are generally highly diffuse, yet small quantities of pathogens may pose a health risk. This study evaluates the ability of TransPlex™ whole transcriptome amplification (WTA) to amplify small quantities of RNA viruses from complex environmental matrices containing background nucleic acids.
  
  
  RESULTS
  DNA extracts from mock drinking water samples containing mixed microbial populations were spiked with small quantities of echovirus type 13 (EV) RNA. Samples were amplified using a Transplex™ WTA kit, and EV-specific quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to quantify target pathogens before and after application of WTA. Samples amplified by WTA demonstrated a decreased limit of detection. The log-linear relationship between serial dilutions was maintained following amplification by WTA.
  
  
  CONCLUSIONS
  WTA is able to increase the quantity of target organism RNA in mixed populations, while maintaining log linearity of amplification across different target concentrations.
  
  
  CONCLUSIONS
  WTA may serve as an effective preamplification step to increase the levels of RNA prior to detection by other molecular methods such as PCR, microarrays and sequencing.
authors.affiliation: Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA, USA.
authors.forename: J K
authors.initials: JK
authors.lastname: Parker
authors.forename: T-Y
authors.initials: TY
authors.lastname: Chang
authors.forename: J S
authors.initials: JS
authors.lastname: Meschke
mesh_terms: D029822:Enterovirus B, Human / Q000235:genetics / Q000302:isolation & purification*; D020869:Gene Expression Profiling / Q000379:methods; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics; D014871:Water Microbiology; D014881:Water Supply
chemical_list: D012367:RNA, Viral