Abstract
Parechovirus A (PeV-A; human parechovirus) causes mild infections and severe diseases such as neonatal sepsis, encephalitis, and cardiomyopathy in young children. Among the 19 types of PeV-A, PeV-A1 is the most common type of infection. We have previously established an immunofluorescence assay for detecting multiple PeV-A types with a polyclonal antibody against the conserved epitope of VP0. Although the polyclonal antibody is useful for PeV-A diagnosis, it could not distinguish the PeV-A genotypes. Thus, the development of a specific monoclonal antibody for identifying the common infection of PeV-A1 would be beneficial in clinical diagnosis practice. In this study, the recombinant full-length PeV-A1 VP0 protein was used in mouse immunization; a total 10 hybridomas were established. After evaluation by immunoblotting and fluorescence assays, six hybridoma clones with monoclonal antibody (mAb) production were confirmed. These mAbs, which specifically recognize viral protein PeV-A1 VP0 without cross-reactivity to PeV-A3, will prove useful in research and PeV-A1 diagnosis.
Highlights
Parechovirus A (PeV-A) is a nonenveloped, single-stranded positive-sense RNA virus
Splenocytes isolated from the mice were used for generating hybridomas that were fused with hypoxanthine aminopterin thymidine (HAT)-sensitive myeloma cells based on the polyethylene glycol (PEG)-mediated cell fusion approach
Ten hybridoma clones were obtained, and monoclonal antibody (mAb) produced by them were harvested for further evaluation
Summary
Parechovirus A (PeV-A) is a nonenveloped, single-stranded positive-sense RNA virus. It belongs to the Picornaviridae, genus parechovirus, comprising parechovirus A and B [1]. The PeV-A genome encodes a single polyprotein that is processed into three structural or capsid-encoding proteins (VP0, VP3, and VP1) and seven nonstructural proteins (2A, 2B, 2C (P2); 3A, 3B, 3C and 3D (P3)) [2]. Hospital-acquired infections in the neonatal department appear to be a critical factor in PeV-A infection [8,9]. An effective method for identifying PeV-A infection in hospitals is important for patient care and transmission prevention
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