Detection of poliovirus, hepatitis A virus and rotavirus from sewage and water samples.
A modified adsorption-elution technique for concentration of enteric viruses from sewage and water samples was developed. The viruses in water were concentrated by negatively charged membrane filtration, eluted with 2.9% tryptose phosphate broth containing 6% glycine pH 9.0, and reconcentrated using centrifugation by a speedVac concentrator. The presence of poliovirus, hepatitis A virus (HAV) RNA, and rotavirus antigen was determined by cell culture isolation, nested polymerase chain reaction (nested PCR), and enzyme-linked immunosorbent assay (ELISA), respectively. A total of 100 sewage and water samples were collected from various sources in congested communities in Bangkok, concentrated and examined for those enteric viruses. Of 20 surface water samples from canals which located near sewage drains, 15% were positive for HAV RNA by nested PCR. Of 48 domestic sewage samples from man-holes of underground sewers, 8% were positive for rotavirus antigen by ELISA. Even though the samples were concentrated 256-2,000 fold, poliovirus was not found by isolation in cell culture.
Kittigul
L
L
Department of Microbiology, Faculty of Public Health, Mahidol University, Bangkok, Thailand. phlkt@mahidol.ac.th
Raengsakulrach
B
B
Siritantikorn
S
S
Kanyok
R
R
Utrarachkij
F
F
Diraphat
P
P
Thirawuth
V
V
Siripanichgon
K
K
Pungchitton
S
S
Chitpirom
K
K
Chaichantanakit
N
N
Vathanophas
K
K
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis; D002460:Cell Line; D002498:Centrifugation; D004797:Enzyme-Linked Immunosorbent Assay; D005374:Filtration; D006507:Hepatovirus / Q000302:isolation & purification*; D006801:Humans; D008253:Macaca mulatta; D017955:Poliovirus / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis; D012401:Rotavirus / Q000276:immunology / Q000302:isolation & purification*; D012722:Sewage / Q000821:virology*; D013785:Thailand; D014407:Tumor Cells, Cultured; D014776:Virus Cultivation; D014871:Water Microbiology
D000956:Antigens, Viral; D012367:RNA, Viral; D012722:Sewage