The aim of this study was to establish a reliable method of virus detection for the diagnosis of critical
enterovirus infections such as acute infective
encephalitis,
encephalomyelitis and
myocarditis. Because histopathological and immunohistochemical analyses of
paraffin-embedded tissues play an important role in recognizing infectious agents in tissue samples, six in-house polyclonal
antibodies raised against three representative enteroviruses using an indirect immunofluorescence assay and immunohistochemistry were examined. This panel of polyclonal
antibodies recognized three serotypes of enterovirus. Two of the polyclonal
antibodies were raised against denatured virus particles from enterovirus A71, one was raised against the recombinant VP1
protein of coxsackievirus B3, and the other for poliovirus type 1 were raised against denatured virus particles, the recombinant VP1
protein and
peptide 2C. Western blot analysis revealed that each of these
antibodies recognized the corresponding
viral antigen and none cross-reacted with non-enteroviruses within the family Picornaviridae. However, all cross-reacted to some extent with the
antigens derived from other serotypes of enterovirus. Indirect immunofluorescence assay and immunohistochemistry revealed that the virus capsid and non-structural
proteins were localized in the cytoplasm of affected culture cells, and skeletal muscles and neurons in neonatal mice experimentally-infected with human enterovirus. The
antibodies also recognized
antigens derived from recent clinical isolates of enterovirus A71, coxsackievirus B3 and poliovirus. In addition, immunohistochemistry revealed that representative
antibodies tested showed the same recognition pattern according to each serotype. Thus, the panel of in-house anti-enterovirus polyclonal
antibodies described herein will be an important tool for the screening and pathological diagnosis for
enterovirus infections, and may be useful for the classification of different enterovirus serotypes, including coxsackieviruses A and B, echoviruses, enterovirus A71 and poliovirus.