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.
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.
Balkin
Helene B
HB
Molecular, Cellular and Biomedical Sciences Department, University of New Hampshire, Durham, NH, USA. hbalkin@unh.edu
Margolin
Aaron B
AB
D000818:Animals; D018929:Cell Culture Techniques / Q000191:economics / Q000379:methods; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D006801:Humans; D008253:Macaca mulatta; D017955:Poliovirus / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D012680:Sensitivity and Specificity; D014773:Virology / Q000191:economics / Q000379:methods*; D014776:Virus Cultivation / Q000191:economics / Q000379:methods; D014871:Water Microbiology