Key result
BGM cells in suspension yielded significantly lower cycle threshold values (average 25.51) compared to conventional monolayers (average 32.17) for the detection of poliovirus.
Absolute Event Rate: 25.51% vs 32.17%
p-value: p=0.000
Using BGM cells in suspension rather than monolayers for ICC/qRT-PCR detection of poliovirus in water samples provides greater sensitivity, requires less hands-on time, and is more cost-effective.
May support more sensitive poliovirus detection in water; leaves open translation to routine surveillance pending validation.
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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Balkin et al. (2010) studied Poliovirus detection in environmental waters. BGM cells in suspension vs. BGM cells in monolayers was evaluated on Cycle threshold (Ct) values by ICC/qRT-PCR (p=0.000). BGM cells in suspension yielded significantly lower cycle threshold values (average 25.51) compared to conventional monolayers (average 32.17) for the detection of poliovirus.
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