Comparison of different concentration methods for the detection of hepatitis A virus and calicivirus from bottled natural mineral waters.
Viral contamination of drinking water is frequently reported as the primary source of gastroenteritis or hepatitis outbreaks. The presence of viruses at low concentration levels in most environmental water poses major analytical problems when determining their concentration. To evaluate the efficiency of different recovery methods of viral RNA from bottled water, a comparison was made of 2 positively and 2 negatively charged membranes that were used for absorbing and releasing HAV virus particles during the filtration of viral spiked bottled water. All the 4 membranes, regardless of charge and pore size, had low level viral recovery. The results show that a considerable number of the virus particles passed through the pores of the membranes instead of being trapped by the electrostatic charges. Two different procedures were then compared using 1.5L polyethylene bottles spiked with 10-fold serial dilutions of HAV and FCV. The first procedure included an ultrafiltration-based method followed by MiniMag RNA extraction, and the second an ultracentrifugation-based method followed by RNA extraction using QIAamp viral RNA mini kit. The ultracentrifugation-based method resulted in a better recovery of HAV and FCV when compared to the ultrafiltration-based method.
Di Pasquale
S
S
Department of Veterinary Public Health and Food Safety, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy. simona.dipasquale@iss.it
Paniconi
M
M
Auricchio
B
B
Orefice
L
L
Schultz
A C
AC
De Medici
D
D
D002139:Caliciviridae / Q000302:isolation & purification*; D005374:Filtration / Q000379:methods; D005516:Food Microbiology; D030041:Hepatitis A virus / Q000302:isolation & purification*; D006801:Humans; D008900:Mineral Waters / Q000821:virology*; D012367:RNA, Viral / Q000302:isolation & purification; D014461:Ultracentrifugation / Q000379:methods; D014462:Ultrafiltration / Q000379:methods; D014773:Virology / Q000379:methods*
D008900:Mineral Waters; D012367:RNA, Viral