Combination of crossflow ultrafiltration, monolithic affinity filtration, and quantitative reverse transcriptase PCR for rapid concentration and quantification of model viruses in water.
We present a rapid and effective adsorption-elution method based on monolithic affinity filtration (MAF) for the concentration and purification of waterborne viruses. The MAF column consists of a hydrolyzed macroporous epoxy-based polymer. High recoveries were achieved by columns for the bacterial virus (bacteriophage) MS2 110 (±19)%, as model organism, as well as for human adenoviruses 42.4 (±3.4)% and murine noroviruses 42.6 (±1.9)%. This new concentration and purification method was combined with crossflow ultrafiltration (CUF). Because of the adsorption of the examined viruses to the macroporous surface of the MAF column at pH 3, concentrated matrix components by CUF can be removed. Bacteriophages MS2 were spiked in tap water and concentrated with the new CUF-MAF concentration method by a volumetric factor of 10(4) within 33 min. Furthermore, the detection limit for quantification of bacteriophage MS2 by quantitative reverse transcriptase PCR (qRT-PCR) could be improved from 79.47 to 0.0056 GU mL(-1) by a factor of 1.4 × 10(4). In a first study, we have shown that this method could also be applied for river water containing naturally MS2 and MS2-like phages.
Chair for Analytical Chemistry and Institute of Hydrochemistry, Technische Universität München, Munich, Germany.
Lu
L
Pei
Martin
M
Rieger
Sandra
S
Lengger
Sonja
S
Ott
Claudia
C
Zawadsky
Nils Marten
NM
Hartmann
Hans-Christoph
HC
Selinka
Andreas
A
Tiehm
Reinhard
R
Niessner
Michael
M
Seidel
D000256:Adenoviridae / Q000235:genetics / Q000302:isolation & purification*; D000818:Animals; D002460:Cell Line; D004867:Equipment Design; D006801:Humans; D017909:Levivirus / Q000235:genetics / Q000302:isolation & purification*; D057230:Limit of Detection; D051379:Mice; D029322:Norovirus / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000295:instrumentation*; D045483:Rivers / Q000382:microbiology*; D014462:Ultrafiltration / Q000295:instrumentation*
D012367:RNA, Viral