Comparison of a reporter assay and immunomagnetic separation real-time reverse transcription-PCR for the detection of enteroviruses in seeded environmental water samples.
Two newly developed protocols for infective virus detection were compared to the plaque assay. An immunomagnetic separation procedure coupled with real-time reverse transcription-PCR of viral nucleic acids was developed to identify intact enteroviral particles, and a reporter cell system responding to viral replication based on fluorescent resonance energy transfer for detection of infectious enteroviruses was tested. Both new procedures detected infective viruses in environmental samples at the same level as the plaque assay.
Hwang
Yu-Chen
YC
Department of Environmental Science, University of California, Riverside, CA 92521, USA.
Leong
Oymon M
OM
Chen
Wilfred
W
Yates
Marylynn V
MV
D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000378:metabolism; D004770:Enterovirus / Q000235:genetics / Q000302:isolation & purification*; D031541:Fluorescence Resonance Energy Transfer; D018189:Immunomagnetic Separation / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D013379:Substrate Specificity; D014764:Viral Proteins / Q000378:metabolism; D014871:Water Microbiology
D014764:Viral Proteins; D003546:Cysteine Endopeptidases; C057400:picornain 2A, Picornavirus