Research and Development - Drinking Water
Title: The Detection of Astrovirus, Enteroviruses, and Adenovirus
type 40 and 41 in Surface Waters Collected and Evaluated by the
Information Collection Rule and an Integrated Cell Culture / Nested
PCR Procedure
Authors: Christopher D. Chapron, Nicola A. Ballester,
Justin H. Fontaine, Christine N. Frades and Aaron B. Margolin
ABSTRACT:
We evaluated the use of an integrated cell culture-reverse transcription
polymerase chain reaction (ICC-RT-PCR) coupled with nested PCR to
detect human astroviruses, enteroviruses and adenovirus type 40
and 41 in surface water samples collected and evaluated by the Information
Collection Rule (ICR) method. The results of the ICC-RT-PCR/Nested
PCR method were compared to those of the Total Culturable Virus
MPN assay (TCVA MPN, the EPA recommended method for monitoring viruses
in surface and finished waters). Twenty-nine ICR surface water samples
were analyzed. Viruses were concentrated by filter adsorption-beef
extract elution and organic flocculation and then evaluated for
viruses by the visualization of cytopathic effect (CPE) in the BGMK
cell line. The ICC-RT-PCR/Nested PCR technique used Caco-2 cells
for the propagation of astrovirus and enteroviruses (ICC step) and
BGMK cells for adenovirus type 40 and 41 as well as for enteroviruses.
Fifteen samples (51.7%) were positive by ICC-RT-PCR/Nested PCR for
astrovirus. Eight of those samples (27.5%) were determined to contain
infectious astrovirus. Seventeen samples (58.6%) were positive for
enteroviruses using the BGMK cell line. Six (27.6%) of those samples
were determined infectious. Fourteen (48.3%) samples were positive
for adenovirus type 40 and 41, eleven (37.9%) of which were determined
to be infectious. Twenty-seven of the twenty-nine (93.1%) samples
were positive for virus, of which, nineteen (68.9%) samples were
positive for infectious virus. Only five of the twenty-nine samples
(17.2%) were positive by the TCVA MPN method. ICC-RT-PCR/Nested
PCR provided an increased sensitivity upon comparison to the TCVA
MPN method.
Published in Applied & Environmental Microbiology, June 2000 |
