Evaluation of ATCC^® Quantitative Synthetic RNA from Zika virus for Diagnostic Development and Validation

Zika virus (ZIKV) is an enveloped, single-stranded RNA arbovirus in the Flaviviridae family, with two major genotypes: African and Asian. It is transmitted primarily by infected Aedes aegypti mosquitoes and, less commonly, through sexual contact or from mother to fetus during pregnancy. While most infections are mild or asymptomatic, ZIKV can cause serious congenital abnormalities like microcephaly and has been linked to Guillain–Barré syndrome in adults.

Because Zika symptoms resemble those of other arboviruses, accurate detection is essential. Traditional virus culture is slow and expensive, while PCR-based methods provide fast, sensitive results but require high-quality reference materials. Meeting this need, ATCC developed a quantitative synthetic analytical reference material (ARM) for ZIKV (ATCC® VR-3252SD™) that contains key genomic regions targeted by widely used molecular detection assays, including prM, E, NS1, NS2B, NS3, NS4B, and NS5. The construct doesn’t contain infectious material and can be handled safely in BSL-1 laboratories, unlike live ZIKV, which requires BSL-2 containment.

The ARM was validated by next-generation sequencing, and RNA copy number was quantified using digital-based PCR. Its performance was evaluated across qRT-PCR assays developed by the CDC, WHO, and Faye et al. (2013) and benchmarked against genomic RNA from Asian (ATCC® VR-1843DQ™) and African (ATCC® VR-1838DQ™) ZIKV lineages. Ten-fold serial dilutions ranging from 50 to 50,000 genome copies per reaction generated a standard curve with R² ≥ 0.999 and an average slope of -3.443. These results confirm that the synthetic ARM provides a reliable ARM for the detection of ZIKV across diverse assay platforms.