J Bacteriol Virol.  2008 Jun;38(2):77-87. 10.4167/jbv.2008.38.2.77.

Integrated Cell Culture-PCR Detection of Enteroviruses and Reoviruses in Water Sources in Gyeonggi-do

• Kim KA
• Kim JC
• Ko HU
• Lee JB
• Kim YS
• Park YB
• Lee MJ
• Kim MG
• Kim JK
• Park EM

• KMID: 1483977
• DOI: http://doi.org/10.4167/jbv.2008.38.2.77

Abstract

The integrated cell culture-PCR (ICC-PCR) method has been suggested as an improved method for detection of viruses in water environments. We tested 57 source waters including finished water samples in Gyeonggi-do for enteric viral contamination using total culturable virus assay (TCVA) using BGMK cells and ICC-PCR. Nineteen of the 57 source water samples (33.3%) exhibited the cytopathic effect (CPE) on BGMK cells and no finished water did exhibited CPE. Nineteen samples (33.3%) of the 57 were positive for reoviruses. For the enteroviruses, only 3 samples (5.3%) of the 57 samples showed positive results. By using ICC-PCR method, 202 flasks from source water samples were positive for enteroviruses and reoviruses. Three samples from source water were positive for both viruses. However, any flasks tested was not co-infected with two types of viruses. While the enteric viral frequencies in TCVA and ICC-PCR were similar, the viral frequency for reoviruses at first passage in two type of method was higher in ICC-PCR (94.7%) than TCVA (56.9%).

Keyword

Integrated cell culture-PCR (ICC-PCR); Total culturable virus assay (TCVA); Enteroviruses; Reoviruses

MeSH Terms

Enterovirus
Viruses
Water
Water

Reference

References

1). 정용석. 수인성 바이러스의 수계환경내 분포실태 및 검색방법론. 지구환경논문집. 9:88–95. 1998.

2). 정현미. 바이러스 수질분석과 정도관리. 지구환경논문 집. 10:55–72. 1999.

3). 지연숙, 이희숙, 남세희, 이기종. 수인성 바이러스 분 석에서 RT-PCR (Reverse transcription Polymerase Chain Reaction)과 IFA (Immunofluorescence Assay)를 이용한 총배양성바이러스 분석법의 보완. 대한환경공학회 추 계학술연구발표회논문집. 1464–1469. 2003.

4). Abbaszadegan M, Stewart P, LeChevallier M. A Strategy for detection of viruses in groundwater by PCR. Appl Environ Microbiol. 65:444–449. 1999.

5). American Public Health Association. Standard Methods for the Examination of Water and Wastewater. 19th ed.American Public Health Association;Washington D. C.: 1995.

6). Berg D, Metcalf TG. Indicators of viruses in water. In. Indicatiors of viruses in water and food. Berg G, editor. (Ed),. Ann Arbor Scinece;Ann Arbor, MI: 1978.

7). Berlin LE, Rorabaugh ML, Heldrich F, Roberts K, Doran T, Modlin JF. Aseptic meningitis in infants <2yr of age: diagnosis and etiology. J Infect Dis. 168:888–892. 1993.

8). Blackmer F, Reynolds KA, Gerba CP, Pepper IL. Use of integrated cell culture-PCR to evaluate the effectiveness of poliovirus inactivation by cholrine. Appl Environ Microbiol. 66:2267–2268. 2000.

9). Chapron CD, Ballester NA, Fontatine 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. 66:2520–2525. 2000.

10). Dahing DR, Wright BA. Optimization of the BGM cell line culture and viral assay procedures for monitoring viruses in the environment. Appl Environ Microbiol. 51:790–812. 1986.

11). Fout GS, Martinson BC, Moyer MW, Dahling DR. A multiplex reverse transcription-PCR method for detection of human enteric viruses in groundwater. Appl Environ Microbiol. 69:3158–3164. 2003.

12). Fraenkel-Conrat H, Kimball P, Levy J. Virology. 2nd ed.Prentice Hall;Englewood Cliffs, NJ: 1988.

13). Grabow WO, Botma KL, de Villiers JC, Clay CG, Erasmus B. Assessment of cell culture and polymerase chain reaction procedures for the detection of polioviruses in wastewater. Bull World Health Organ. 77:973–980. 1999.

14). Greening GE, Hewitt J, Lewis GD. Evaluation of integrated cell culture-PCR (C-PCR) for virological analysis of environmental samples. J Appl Microbiol. 93:745–750. 2002.

15). Grimm AC, Cashdollar JL, Williams FP, Fout GS. Development of an astrovirus RT-PCR detection assay for use with conventional, real-time, and integrated cell culture/RT-PCR. Can J Microbiol. 50:269–278. 2004.

16). Gurrman-Bass N, Catalano-Sherman J. Effects of humic materials on virus recovery from water. Appl Environ Microbiol. 49:1260–1264. 1985.

17). Harris GD, Adams VD, Sorensen DL, Curtis MS. Ultraviolet inactivation of selected bacteria and viruses with photo-reactivation of the bacteria. Water Res. 21:687–692. 1987.

18). Irving LG, Smith FA. One-year survey of enteroviruses, adenoviruses, and reoviruses isolated from effluent at an activated-sludge purification plant. Appl Environ Microbiol. 41:51–59. 1981.

19). Johansson PJ, Sveger T, Ahlfors K, Ekstrand J, Svensson L. Reovirus type 1 associated with meningitis. Scand J Infect Dis. 28:117–120. 1996.

20). LaBelle RL, Gerba CP. Influence of estuarine sediment on virus survival under field conditions. Appl Environ Microbiol. 39:749–755. 1980.

21). Lee CH, Lee SH, Han ER, Kim SJ. Use of cell culture-PCR assay based on combination of A549 and BGMK cell lines and molecular identification as a tool to monitor infectious adenoviruses and enteroviruses in river water. Appl Environ Microbiol. 70:6695–6705. 2004.

22). Leparc I, Aymard M, Fuchs F. Acute, chronic and persistent enterovirus and poliovirus infection: detection of viral genome by seminested PCR amplification in culture-negative samples. Mol Cell Probes. 8:487–495. 1994.

23). Lerner AM, Cherry JD, Klein JO, Finland M. Infection with reoviruses. N Engl J Med. 267:947–952. 1962.

24). Melnick JL. Enteroviruses: polioviruses, coxsackieviruses, echoviruses and newer enteroviruses. pp. p. 549–605. In. Virology. Fields BN, editor. (Ed),. Raven Press;New York: 1990.

25). Minor PD, Morgan-Capner P, Schild GC. Enteroviruses. pp. p. 417–439. In. Principles and practice of clinical virology. 3rd ed. Zucherman AJ, Pattison JR, editors. (Ed),. Wiley & Sons Ltd.;Chichester: 1994.

26). Morris R, Waite WM. Evaluation of procedures for recovery of viruses from water. II. Detection systems. Water Res. 14:795–798. 1980.

27). Muscillo M, Aulicino FA, Patti AM, Orsini P, Volterra L, Fara GM. Molecular techniques for the identification of enteric viruses in marine waters. Water Res. 28:1–7. 1994.

28). Muscillo M, Carducci A, La Rosa G, Cantiani L, Marianelli C. Enteric virus detection in Adriatic seawater by cell culture, polymerase chain reaction and polyacrylamide gel electrophoresis. Water Res. 31:1980–1984. 1997.

29). Muscillo M, La Rosa G, Marianelli C, Zaniratti S, Capobianchi MR, Cantiani L, Carducci A. A new RT-PCR method for the identification of reoviruses in seawater samples. Water Res. 35:548–556. 2001.

30). Puig M, Jofre J, Lucena F, Allard A, Wadell G, Girones R. Detection of adenoviruses and enteroviruses in polluted waters by nested PCR amplification. Appl Environ Microbiol. 60:2963–2970. 1994.

31). Reynolds KA. Integrated cell culture/PCR for detection of enteric viruses in environmental samples. Methods Mol Biol. 268:69–78. 2004.

32). Reynolds KA, Gerba CP, Abbaszadegan M, Pepper LL. ICC/PCR detection of enteroviruses and hepatitis A virus in environmental samples. Can J Microbiol. 47:153–157. 2001.

33). Reynolds KA, Gerba CP, Pepper IL. Detection of infectious enteroviruses by an integrated cell culture-PCR procedure. Appl Environ Microbiol. 62:1424–1427. 1996.

34). Rigotto C, Sincero TCM, Simões CMO, Barardi CRM. Detection of adenoviruses in shellfish by means of conventional-PCR, nested-PCR, and integrated cell culture PCR (ICC/PCR). Water Res. 39:297–304. 2005.

35). Rusin P, Enriquez C, Johnson D, Gerba C. Enteric viruses. pp. p. 472–484. In. Environmental microbiology. 1st ed. Maier RM, Pepper IL, Gerba CP, editors. (Ed),. Academic Press;San Diego, CA: 2000.

36). Sabin AB. Reoviruses. a new group of respiratory and enteric viruses formerly classified as ECHO type 10 is described. Science. 130:1387–1389. 1959.

37). Sellwood J, Dadswell JV, Slade JS. Viruses in sewage as and indicator of their presence in the community. J Hyg. 86:217–225. 1981.

38). Shieh YS, Wait D, Tai L, Sobsey MD. Methods to remove inhibitors in sewage and other fecal wastes for enterovirus detection by the polymerase chain reaction. J Virol Methods. 54:51–66. 1995.

39). Sobsey MD. Methods for recovering viruses from shellfish, seawater, and sediments. pp. p. 77–108. In. Methods for recovering viruses form the environment. Berg G, editor. (Ed),. CRC Press, Inc.;Boca Raton, FL: 1987.

40). Sobsey MD, Hickey AR. Effects of humic and fulvic acids on poliovirus concentration from water by microporous filtration. Appl Environ Microbiol. 49:259–264. 1985.

41). Spinner ML, Di Giovanni GD. Detection and identification of mammalian reoviruses in surface water by combined cell culture and reverse transciption-PCR. Appl Environ Microbiol. 67:3016–3020. 2001.

42). Tyler KL. Mammalian reoviruses. Fields virology. 4th ed. Knipe DM, Fields BN, Howley PM, Griffin DE, editors. (Ed),. Lippincott Williams & Wilkins;Philadelphia, PA: 2001.

43). United States Environmental Protection Agency. ICR Microbial Laboratory Manual: Office of Research and Development. EPA/600.R-95/178.1996.

44). Vantarakis AC, Parapetropoulou M. Detection of entero-viruses and adonoviruses in coastal waters of SW Greece by nested polymerase chain reaction. Water Res. 32:2365–2372. 1998.