A Comparative study of ICC-RT-PCR and Cell Culture methods for detection of Enteroviruses in Sewage
Subject Areas : Diagnostic Microbiology
Mohammad Kargar
1
(Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran)
Sara Sadeghipour
2
(Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran)
Hamideh Tabatabaee
3
(Department of Virology, School of Public Health Tehran University, Tehran, Iran.)
Rakhshandeh Nategh
4
(Department of Virology, School of Public Health Tehran University, Tehran, Iran)
Keywords: ICC-RT-PCR, Enteroviruses, Sewage, Cell- Culture,
Abstract :
Background and objectives: Enteroviruses are one of the most important enteric viruses which produce a broad range of human diseases. But being time consuming and also the necessity to perform confirmed tests for Enteroviruses detection, using molecular direct methods have been recently considered. The aim of this research is evaluation of ICC-RT-PCR and cell culture methods for detection of enteroviruses in sewage system. Materials and methods: In this research 63 specimens were provided through Grab sample method which concentrated by pellet & 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 360C by means of Pan E.V primers and afterwards Pan P.V Primers along with specific sabin primers, test of RT-PCR was carried out on the cell culture specimens. Results: Out of all 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. Conclusion: Statistical analysis indicated that there is a significant relationship (p= 0.05) between cell culture and ICC-RT-PCR methods for isolating enteroviruses. Further the sensitivity of ICC-RTPCR method to detect enteroviruses less than 0.01 TCID50 was evaluated, which can indicated the acceptability and sensitivity of this method for detection of enteroviruses in sewages.