Isolation of polioviruses and other enteroviruses in south Greece between 1994 and 1998

Abstract

During the five‐year period between 1994 and 1998, a total of 217 clinical samples were assessed for the isolation of enteroviruses at the Enterovirus Reference Centre for South Greece. Fourteen enterovirus strains belonging to different serotypes were isolated. These field strains were detected by cell culture in appropriate cell lines. They were subsequently identified by neutralizing antibodies with the LBM (Lim‐Benyesh Melnick) mixed antisera pools up to 1995 and RIVM (National Institute of Public Health and the Environment, Bilthoven, The Netherlands) pools from 1996 onwards. The isolated viruses included two strains of poliovirus type 2 Sabin‐like, three strains of poliovirus type 1 non‐Sabin‐like, one Coxsackie B2 (CBV2) strain, one Coxsackie B5 (CBV5) strain, one Echo 5 (ECV5) strain, one Echo 7 (ECV7) strain, three Coxsackie A16 (CAV16) strains, and two currently enteroviral strains unidentified by RIVM pools. Reverse transcription‐polymerase chain reaction (RT‐PCR) using poliovirus‐specific primers or poliovirus type‐specific primers and enterovirus specific primers from the highly conserved 5′‐UTR, the latter followed by RFLP, was also applied in 6 clinical isolates (3 strains of poliovirus type 1 non‐Sabin‐like, 1 polio type 2 Sabin‐like, and 2 non‐identified by RIVM pools enteroviruses). The advantages and the drawbacks of these assays against the conventional ones are discussed here. The isolations and the subsequent identification of the strains were carried out from fecal samples of clinical cases that included hand‐foot‐and‐mouth disease, meningitis, and acute flaccid paralysis. The reappearance of non‐Sabin‐like poliovirus strains in Greece in 1996 after 14 years is considered to have an important medical and clinical value. J. Clin. Lab. Anal. 14:157–163, 2000. © 2000 Wiley‐Liss, Inc.