RAPID SUBGROUPING OF NONPOLIO ENTEROVIRUS ASSOCIATED WITH ASEPTIC-MENINGITIS BY RFLP (RESTRICTION-FRAGMENT-LENGTH-POLYMORPHISM) ASSAY

In Korea, there was a big outbreak of Aseptic Meningitis due to entero virus infection in 1993, Since virus isolation and neutralizing tests are too laborious and time-consuming for the detection of enterovirus from clinical specimen, we have developed a new molecular identificati on method for rapid subgrouping of isolates from patients with aseptic meningitis. For the rapid subgrouping of isolates, RT-PCR (Reverse Tr anscription-Polymerase Chain Reaction) and RFLP (Restriction Fragment Length Polymorphism) assays were used. We have selected two oligonucle otide primers from the conserved 5'-UTR/VP2 and VP1 regions. A 652 bp (base pair) product was amplified from the 5'-UTR/VP2 region of refere nce viruses and the isolates. For the subgrouping of the isolates by R FLP assay, we have used 12 reference viruses (Echovirus, E6, E9, E11, E12, Cossackievirus, CB1, CB3, CB4, CB5, Cossackievirus, CA9, CA16, CA 21, CA24), which are the common viral agents associated with aseptic m eningitis. By using subgroup-specific restriction enzymes BsmAI, HinP1 I, and PleI, the isolates were classified into Echovirus subgroups. We have also shown that subgrouping of the isolates by RFLP assay based on the VP1 region is possible.