الفهرس | Only 14 pages are availabe for public view |
Abstract A total of 300 stool samples were collected from 300 infants (6 months to 5 years old) geographically distribute among Egypt, between the period of May to October 2004, the samples were cultured on RD cell line. After 2 blind passages. Out of 300 cultured samples, 63 (21 %) were positive for CPE where as 237 (79%) negative. An additional passage was carried out to exclude the negative samples and to increase the virus titer of the positive isolates for subjected to titration. Regarding the months of the sample collection, the highest isolation rate was found in July (23.63%) and August (25%) whereas the lowest rate was in May (17.5%) and October (17.77). The positive isolates (63 in numbers) were typed by neutralization test utilizing the pp (polio pool) 1,2 and 3; CP (coxsackie pool, B 1- 6) and seven pools (A-G) against coxsackie A9 and 20 Echo. Out of 63 isolates, 13 isolates (21%) were typed as Coxsackie B, 28 (44%) were identified as non polio enteroviruses (NPEV) and 22 (35%) were Echoviruses and sub classified into Echo 1, 3, 4, 6, 9, 11, 13, 25, 27, 29 and 30. Echoviruses type 3,6,11 and 13 predominating. The RNA extracted from positive isolates and used in RT-PCR, amplification reaction carried out with specific primers selected from the highly conserved parts of the 5’ non-coding region of the EV genome. The amplification reaction was carried out and the expected and correct molecular size of the PCR product is 114 bp. All the tested samples revealed positive amplification in the employed RT-PCR assay confirming the successful isolation of Enteroviruses when electrophoresis of the amplified products on 10 % polyacrylamide gel. Also probe hybridization assay was carried out using Enterovirus group probe (EV/5’UT) capable of forming stable hybrids with all enteroviruses RNAs. All the tested samples revealed positive dot blot in hybridization assay confirming the successful isolation of Enteroviruses. This study report for the first time HEV isolates and show the most predominant Echoviruses 3,6,11 and 13. |