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

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

[Assessment of different methods to remove organic inhibitors in order to create a sensitive method for direct surveillance of enteroviral infectious diseases in sewage specimens]

Enteroviruses in sewage are considered among the most sensitive indicators for virus circulation in society. These are mainly detected by sensitive cell cultures, however, since it is time consuming, molecular direct methods have also been considered as sensitive techniques. This study accomplished to assess different methods of removing organic inhibitors of sewage in order to detect Enteroviruses with RT-PCR. For this cross sectional study, 63 sewage specimens of Tehran were prepared with Grab sample method and concentrated with Pellet and Two-Phase methods and cultured in RD and Hep-2 cells. Then, with 12 different methods, removing organic inhibitors in sewage with RT-PCR method was assessed. Of investigated methods, we succeeded to isolate all of expected viruses only with the use of ICC-RT-PCR method. With respect to 0.01 TCID50 sensitivity of ICC-RT-PCR method, further studies are strongly recommended in order to confirm the utility of this method as one of the most sensitive methods of detecting Enteroviruses

Congenital pouch colon with imperforate anus; a report of 3 cases

Congenital pouch colon [CPC] is an unusal condition in imperforate anus [IA] usually of high variety this malfotmation is mostly reported from India, where it comprises about 7% of all anorectal malformations. In this anomaly the normal colon is very short or absent and is replaced by a pouch like dilatation that usually communicates through a fistula with genitourinary tract or persistent cloaca. We are reporting on three cases of newborns [2 females, and 1 male] with this anomaly diagnosed during September 2001 to September 2004 These three cases comprise less than 3% of all cases of IA during this time period. The two girls had anorectal agenesia, short segment of colon and a pouch with vaginal fistula and persistent cloaca [Type II]. Colon in the third case was replaced by a dilated pouch that communicated with a wide fistula to the bladder [Type I]. This is very important for pediatric surgeons to be aware of the features of this condition to allow for proper diagnosis and surgical management