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

• Main Authors: M Kargar (Author), S Sadeghipour (Author), B Zarei Mahmoudabadi (Author), R Nategh (Author)
• Format: Book
• Published: Tehran University of Medical Sciences, 2009-09-01T00:00:00Z.
• Subjects: article
• Online Access: Connect to this object online.
• Notes: 2251-6085; 2251-6093
• Keywords: Enteroviruses; Sewage; ICC-RT-PCR; Iran; Public aspects of medicine; RA1-1270
• Source: Iranian Journal of Public Health, Vol 38, Iss 3 (2009)

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.