Establishment of a panel of in‐house polyclonal antibodies for the diagnosis of enterovirus infections

Abstract

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.

Neuropathogenicity of Two Saffold Virus Type 3 Isolates in Mouse Models

“…Highly neurotropic viruses such as poliovirus, enterovirus 71, and neurovirulent flaviviruses, infect the CNS and cause viral meningitis or encephalomyelitis, which is characterized by neuronal damage and perivascular inflammation [67][68][69][70]. The preferential target of these neurotropic viruses is the large pyramidal neurons in the human CNS (and in that of animal models) [38,71,72]. In this study, the two SAFV-3 strains only infected neural progenitor cells and glial cells around the lateral ventricles and cerebellum in both neonatal and young mice.…”

Evaluation of Laboratory Application of Camelid Sera Containing Heavy-Chain Polyclonal Antibody Against Recombinant Cytotoxic T-Lymphocyte-Associated Protein-4

“…(34) The result of applying six different rabbit pAbs in WB and IHC, showed these antibodies can screen and determine various enterovirus antigens and be sensitive for some serotype classification. (35) In another study, the activity of purified pAbs was evaluated and confirmed through different laboratory methods. Their results revealed that the raised rabbit-specific pAbs against human extracellular signal-regulated kinase 8 were able to perform as a sensitive antibody for cancer case studies.…”

Congenital Echovirus 11 Infection in a Neonate

“…Immunohistochemistry using human enterovirus A, B, and C antibodies revealed that only enterovirus B antigen was detected in the liver and adrenal gland in accordance with broad hemorrhagic necrosis (Fig. 2), 9 but not in the placenta. However, enterovirus RNA was extracted using viral RT-PCR not only in the liver (631 copies/cell) and adrenal gland (3510 copies/cell) but also in the placenta (2.2 copies/cell) and spleen (426 copies/cell) (Table 2).…”