Computational Design of a Synthetic Molecular Standard for Human Parechovirus 3

Human parechovirus 3 (HPeV3) has been increasingly identified in cases of aseptic meningitis among neonates and young infants less than 1 year of age and is associated with paralysis, viral-like sepsis, central nervous system (CNS) infection, and sudden death. Because these clinical manifestations are similar to those associated with enterovirus infections, HPeV3 infections are often misdiagnosed, which in turn results in poor patient outcome. Therefore, molecular detection assays that provide a rapid and accurate diagnosis of HPeV3 are critical for ensuring prompt and appropriate treaent. Due to its sensitivity and quick turnaround time, the preferred method for the detection of RNA viruses is quantitative reverse-transcription PCR (qRT-PCR), which relies on the generation of a standard curve that is prepared using a quantitative viral RNA standard. To this end, we have designed, developed, and quantified a synthetic molecular standard for HPeV3 that is compliant with ISO 13485. This preparation is supported by stringent quality control analyses to ensure product identity, stability, and functionality with molecular applications, making it an ideal control for assessing assay performance and ensuring accurate and reproducible results. In the following proof-of-concept study, the HPeV3 synthetic molecular standard was quantified using Droplet Digital PCR (ddPCR; Bio-Rad) and validated via qRT-PCR using published primers.