Literature DB >> 20974002

Detection of poliovirus by ICC/qPCR in concentrated water samples has greater sensitivity and is less costly using BGM cells in suspension as compared to monolayers.

Helene B Balkin1, Aaron B Margolin.   

Abstract

The integrated cell culture quantitative reverse transcriptase PCR (ICC/qRT-PCR) method is used in our lab to detect enteroviruses in environmental waters. Typically we utilize monolayers of 3 cell lines; buffalo green monkey kidney (BGM), human colonic carcinoma (CACO-2) and African rhesus monkey kidney (MA104) with the intent of providing one or more permissive hosts to a wide range of enteroviruses. In this study the BGM cell line was used to compare poliovirus infectivity in conventional monolayer cultures to BGM cells in suspensions. Propagated virus was subsequently amplified by qRT-PCR. Our PCR data showed lower cycle threshold (Ct) values in the suspensions which corresponded to a higher rate of infectivity than that observed in the monolayers. The difference in Ct values was determined statistically significant by One-way ANOVA (0.000). Infecting BGM cells in suspensions required less hands-on time, less chance of contamination and was more cost effective than utilizing the conventional monolayer technique.

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Year:  2010        PMID: 20974002      PMCID: PMC2978159          DOI: 10.1186/1743-422X-7-282

Source DB:  PubMed          Journal:  Virol J        ISSN: 1743-422X            Impact factor:   4.099


  9 in total

1.  Fortified sera and their use in environmental virology.

Authors:  J L Hoyt; A B Margolin
Journal:  Appl Environ Microbiol       Date:  2000-05       Impact factor: 4.792

2.  Organic flocculation: an efficient second-step concentration method for the detection of viruses in tap water.

Authors:  E Katzenelson; B Fattal; T Hostovesky
Journal:  Appl Environ Microbiol       Date:  1976-10       Impact factor: 4.792

3.  PCR detection of pathogenic viruses in southern California urban rivers.

Authors:  S C Jiang; W Chu
Journal:  J Appl Microbiol       Date:  2004       Impact factor: 3.772

4.  Real-time PCR quantification of human adenoviruses in urban rivers indicates genome prevalence but low infectivity.

Authors:  Samuel Choi; Sunny C Jiang
Journal:  Appl Environ Microbiol       Date:  2005-11       Impact factor: 4.792

5.  Suitability of infection of cells in suspension for detection of herpes simplex virus.

Authors:  G Luker; C Chow; D F Richards; F B Johnson
Journal:  J Clin Microbiol       Date:  1991-07       Impact factor: 5.948

6.  Optimization of the BGM cell line culture and viral assay procedures for monitoring viruses in the environment.

Authors:  D R Dahling; B A Wright
Journal:  Appl Environ Microbiol       Date:  1986-04       Impact factor: 4.792

7.  Development of a shaker culture of Buffalo green monkey kidney cells: potential use for detection of enteroviruses.

Authors:  G Goldstein; L E Guskey
Journal:  Appl Environ Microbiol       Date:  1982-08       Impact factor: 4.792

8.  Detection of infectious enteroviruses by an integrated cell culture-PCR procedure.

Authors:  K A Reynolds; C P Gerba; I L Pepper
Journal:  Appl Environ Microbiol       Date:  1996-04       Impact factor: 4.792

Review 9.  Pathogenic human viruses in coastal waters.

Authors:  Dale W Griffin; Kim A Donaldson; John H Paul; Joan B Rose
Journal:  Clin Microbiol Rev       Date:  2003-01       Impact factor: 26.132

  9 in total
  2 in total

Review 1.  Methods to detect infectious human enteric viruses in environmental water samples.

Authors:  Ibrahim Ahmed Hamza; Lars Jurzik; Klaus Überla; Michael Wilhelm
Journal:  Int J Hyg Environ Health       Date:  2011-09-15       Impact factor: 5.840

2.  Monitoring of adenovirus serotypes in environmental samples by combined PCR and melting point analyses.

Authors:  Nils Marten Hartmann; Melanie Dartscht; Regine Szewzyk; Hans-Christoph Selinka
Journal:  Virol J       Date:  2013-06-10       Impact factor: 4.099

  2 in total

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