Comparison of Integrated Cell Culture -RT-PCR and Cell Culture Methods for detection of Enteroviruses

ABSTRACT

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. 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[degree sign] 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. Out of 63 collected specimens, enteroviruses were isolated from 33 specimens [52.38%] and 41[65.01%] specimens which utilized cell-culture and ICC-RT-PCR methods respectively. Polioviruses were also isolated from 6 specimens. 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 enteroviruses less than 0.01 TCID 50 was evaluated, which indicated that this method is acceptable and sensitive enough to detect enteroviruses in sewage