The environmental water samples assayed by total culturable virus assay (TCVA) were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and integrated cell culture-PCR (ICC-PCR) method for enteroviruses and reovirus since they are the usually detected virus groups by culture assays. The detection sensitivities of the TCVA, RT-PCR, and ICC-PCR were compared and the overall reliability of the detection was analyzed to confirm environmental samples for enteric viruses. A total of eight samples from different areas was analyzed by performing TCVA, RT-PCR, and ICC-PCR. Virus concentrations in surface water samples ranged from 1.03 to 47.3 most probable numbers of infectious units (MPN) per 100 liters. When primers specific for both enteroviruses and reoviruses were used in both RT-PCR and ICC-PCR, all the samples (100%) were positive for the viruses. Reoviruses were the most frequently detected ones among the samples. According to the sequence results of enteroviruses, five of the samples were contaminated by coxsackievirus type B3, and the rest by coxsackievirus type B6, echovirus type 30, or vaccine strain poliovirus type 3. It was observed that both enteroviruses and reoviruses were detected concurrently in all CPE-positive environmental water samples.
Abbaszadegan M, Stewart P, LeChevallier M: A strategy for detection of viruses in groundwater by PCR. Appl Environ Microbiol 65: 444-449, 1999
Beller M, Ellis A, Lee SH, Drebot MA, Jenkerson SA, Funk E, Sobsey MD, Simmons OD 3rd, Monroe SS, Ando T, Noel J, Petric M, Middaugh JP, Spika JS: Water-related disease surveillance annual summary 1982. HHS publication No 83-85, 1983.
Parshionikar SU, Willian ST, Fout GS, Robbins DE, Seys SA, Cassady JD, Harris R: Waterborne outbreak of gastroenteritis associated with a norovirus. Appl Environ MicrobioI 69(9): 5263-5268, 2003
Schwab KJ, Leon RD, Sobsey MD: Concentration and purification of beef extract mock eluates from water samples for the detection of enteroviruses, hepatitis A virus, and Norwalk virus by reverse transcription-PCR. Appl Environ MicrobioI 61(2): 531-537, 1995
Steinhauer DA, Holland JJ: Rapid evolution of RNA viruses. Annu Rev Microbiol 41: 409-433, 1987
Cincinnati O, Sally K, Wallace W: The effect of chlorine in water on enteric viruses. Amer Jour Physiol 48: 1323-1334, 1998
Rames M, Sabin B: Role in summer diarrheal disease of infancy and early childhood. J Amer Med Assoc 167: 147-156, 1958
Schwab KJ, Leon RD, Sobsey MD, Moore BE: Survival of human immunodeficiency virus (HIV), HIVinfected lymphocytes, and poliovirus in water. Appl Environ MicrobioI 59(5): 1437-1443, 1995
Spinner ML, Giovanni GDD: Detection and identification of mammalian reoviruses in surface water by combined cell culture and reverse transcription-PCR. Appl Environ MicrobioI 67(7): 3016-3020, 2001
Craun GF: Methods for the investigation and prevention of waterborne disease outbreaks. Appl Environ MicrobioI 61(2): 531-537, 1995
Fout GS, Martinson BC, Moyer MW, Dahling DR: A multiplex reverse transcription-PCR method for detection of human enteric viruses in groundwater. Appl Environ MicrobioI 69(6): 3158-3164, 2003
Alicia D, Anderson, Annette G: A Waterborne Outbreak of Norwalk-Like virus. J Infect Dis 187: 303, 2003
Song KJ, Kang BC, Lee YE, Baek LJ, Lee YJ, Song JW: Isolation and characterization of reovirus in Korea. J Korean Soc Virol 29: 65-74, 1999
Fout GS, Schaefer FWIll, Messer JW, Dahling DR, Stetler RE: Detection of infectious enteroviruses by an integrated cell culture-PCR procedure. Appl Environ MicrobioI 62(4): 1424-1427, 1996
Tani N, Dohi Y, Kurumatani N, Yonemasu K: Seasonal distribution of adenoviruses, enteroviruses and reoviruses in urban river water. Microbiol Immunol 39(8): 577-580, 1995
Abbaszadegan M, Huber MS, Gerba CP, Peper IL: Detection of viruses in groundwater with the polyme-rase chain Reaction. Appl Environ Microbiol 59: 1318-1324, 1993.
Ridinger DN, Spendlove RS, Barnett BB, George DB, Roth JC: Evaluation of cell lines and immuno-fluorescence and plaque assay procedures for quanti-fying reoviruses in sewage. Appl Environ Microbiol 43(4): 740-746,1982.
Allard A, AIbinsson B, Wadell G: Detection of adenoviruses in stools from healthy persons and patients with diarrhea by two-step polymerase chain reaction. J Med Virol 37: 149-157, 1992
Cho HB, Lee SH, Cho JC, Kim SJ: Detection of adenoviruses and enteroviruses in tap water and river water by reverse transcription multiplex PCR. Can J Microbiol 46: 417-424, 2000
Greening GE, Hewitt J, Lewis GD: Evaluation of integrated cell culture-PCR (ICC-PCR) for virological analysis of environmental samples. Appl Environ MicrobioI 93(5): 745-750, 2002
Lee HK, Jeong YS: Comparison of Total Culturable Virus Assay and Multiplex Integrated Cell Culture-PCR for Reliability of Waterborne Virus Detection. Appl Environ Microbiol 70: 3632-3636,2004.
Matsuura K, Hasegawa S, Nakayama T, Morita O, Uetake H: Viral pollution of the rivers in Toyama City. Microbiol ImmunoI 28(5): 575-588, 1984
Oberste MS, Maher K, Flemister MR, Marchetti G, Kilpatrick DR, Pallansch MA: Comparison of classic and molecular approaches for the identification of untypeable enteroviruses. J Clin Microbiol 38(3): 1170-1174, 2000.
Reynolds KA, Gerba CP, Pepper IL: Detection of infectious enteroviruses by an integrated cell culture-PCR procedure. Appl Environ MicrobioI 62(4): 1424-1427, 1996
Richards GP: Limitations of molecular biological techniques for assessing the virological safety of foods. J Food Prot 62(6): 691-697, 1999
Chapron CD, Ballester NA, Fontaine JH, Frades CN, Margolin AB: Detection of astroviruses, enteroviruses, and adenovirus types 40 and 41 in surface waters collected and evaluated by the information collection rule and an integrated cell culture-nested PCR procedure. Appl Environ Microbiol 66(6): 2520-2525, 2000