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Amplification of viral RNA from drinking water using TransPlex™ whole-transcriptome amplification.
J K Parker1, T-Y Chang, J S Meschke
1Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA, USA.
Journal of Applied Microbiology
|April 12, 2011
Summary
Whole transcriptome amplification (WTA) effectively amplifies low-quantity RNA viruses from environmental samples. This method enhances RNA levels for improved pathogen detection using molecular techniques like PCR.
Area of Science:
- Environmental Virology
- Molecular Biology
- Nucleic Acid Amplification
Background:
- Viral pathogens in environmental media are often diffuse, posing health risks even in small quantities.
- Detecting low-abundance RNA viruses in complex environmental matrices is challenging due to background nucleic acids.
Purpose of the Study:
- To evaluate the efficacy of TransPlex™ whole transcriptome amplification (WTA) for amplifying small quantities of RNA viruses.
- To assess WTA's ability to improve the detection of viral RNA in environmental samples.
Main Methods:
- Mock drinking water samples were spiked with echovirus type 13 (EV) RNA.
- TransPlex™ WTA kit was used for RNA amplification.
- EV-specific quantitative reverse transcription polymerase chain reaction (qRT-PCR) was employed for quantification before and after WTA.
Main Results:
- Whole transcriptome amplification (WTA) decreased the limit of detection for target viral RNA.
- The log-linear relationship between serial dilutions of viral RNA was preserved after WTA.
- WTA successfully increased the quantity of target RNA in mixed microbial populations.
Conclusions:
- Whole transcriptome amplification (WTA) is effective for increasing RNA quantity from environmental samples.
- WTA serves as a valuable preamplification step for enhancing RNA levels before downstream molecular detection methods.
- This technique improves the detection sensitivity of RNA viruses in complex environmental matrices.
