S-H Lee1, C Lee, K W Lee, H B Cho, S-J Kim. 1. School of Biological Sciences, College of Natural Sciences, Seoul National University, Seoul, South Korea.
Abstract
AIMS: To evaluate an integrated cell culture (ICC)-multiplex-nested PCR using the buffalo green monkey kidney (BGMK) cells for the simultaneous detection of both enteroviruses and adenoviruses in surface water and tap water samples and optimize the procedure for more sensitive detection of virus showing no apparent cytopathic effect (CPE). METHODS AND RESULTS: A total of 69 surface water and 50 tap water samples were analysed by the ICC-multiplex-nested PCR. All the PCRs were performed five times with a cell lysate from each flask after at least 2 weeks incubation. Forty-six surface water samples (66.7%) and 23 tap water samples (46.0%) exhibited CPE by the cell culture method. By using an ICC-multiplex-nested PCR, 53 surface water samples (76.8%) and 29 tap water samples (58.0%) were determined as containing infectious enteric viral particles. CONCLUSIONS: An ICC-PCR method with a long incubation time using BGMK cells enables the simultaneous detection of enteroviruses and adenoviruses from environmental water samples, including tap water, even with low numbers of viruses. SIGNIFICANCE AND IMPACT OF THE STUDY: A method capable of detecting small numbers of viral particles is necessary.
AIMS: To evaluate an integrated cell culture (ICC)-multiplex-nested PCR using the buffalo green monkey kidney (BGMK) cells for the simultaneous detection of both enteroviruses and adenoviruses in surface water and tap water samples and optimize the procedure for more sensitive detection of virus showing no apparent cytopathic effect (CPE). METHODS AND RESULTS: A total of 69 surface water and 50 tap water samples were analysed by the ICC-multiplex-nested PCR. All the PCRs were performed five times with a cell lysate from each flask after at least 2 weeks incubation. Forty-six surface water samples (66.7%) and 23 tap water samples (46.0%) exhibited CPE by the cell culture method. By using an ICC-multiplex-nested PCR, 53 surface water samples (76.8%) and 29 tap water samples (58.0%) were determined as containing infectious enteric viral particles. CONCLUSIONS: An ICC-PCR method with a long incubation time using BGMK cells enables the simultaneous detection of enteroviruses and adenoviruses from environmental water samples, including tap water, even with low numbers of viruses. SIGNIFICANCE AND IMPACT OF THE STUDY: A method capable of detecting small numbers of viral particles is necessary.
Authors: Andréia Dalla Vecchia; Caroline Rigotto; Rodrigo Staggemeier; Mayra Cristina Soliman; Fernanda Gil de Souza; Andreia Henzel; Eliane Lemos Santos; Carlos Augusto do Nascimento; Daniela Muller de Quevedo; Juliane Deise Fleck; Larissa Schemes Heinzelmann; Sabrina Esteves de Matos Almeida; Fernando Rosado Spilki Journal: Environ Sci Pollut Res Int Date: 2015-02-05 Impact factor: 4.223
Authors: Maryse Iris Sedji; Mihayl Varbanov; Marie Meo; Marius Colin; Laurence Mathieu; Isabelle Bertrand Journal: Environ Sci Pollut Res Int Date: 2018-08-30 Impact factor: 4.223