Comparison of Integrated Cell Culture -RT-PCR & Cell Culture Methods for Detection of Enteroviruses

Abstract

Background: Generally, sewage exposed water could be potentially contaminated with enteroviruses. For this reason, en­terovirus isolation from sewage specimens is one of the most sensitive indicators for virus circulation in the population. We evalu­ated the ICC-RT-PCR and cell culture methods for detection of enteroviruses in Tehran sewage system.
Methods: This research utilized 63 specimens provided through Grab sample method to concentrate by two-phase method and cultured in RD and HEp-2 cells, respectively. All specimens then were inoculated using sensitive cell cultures of RD and HEp-2. After 24 hours incubation at 36 ˚C by means of Pan E.V primers and afterwards Pan P.V Primers along with spe­cific sabin primers, RT-PCR was carried out on the cell culture specimens. Data were analyzed using SPSS Software (SPSS for and ANOVA test as well as Chi-square test.
Results: Out of 63 collected specimens, enteroviruses were isolated from 33 specimens (52.38%) and 41(65.01%) specimens which utilized cell-culture & ICC-RT-PCR methods respectively. Polioviruses were also isolated from 6 specimens.
Conclusions: Statistical analysis indicated that there was a significant relationship (0.05 level) between cell culture and ICC-RT-PCR methods to isolate enteroviruses. Further the sensitivity of ICC-RT-PCR method to detect en-teroviruses less than 0.01 TCID50 was evaluated, which indicated that this method is acceptable and sensitive enough to detect enteroviruses in sewage.