Key result
A novel multiplex PV2-specific real-time quantitative PCR assay identified 98.3% of available Sabin-2 sequences in silico and directly detected PV2 RNA in 183 wastewater samples.
Why the study?
Increasing circulating vaccine-derived poliovirus outbreaks require improved surveillance tools, as the current standard protocol is lengthy, labor-intensive, and inefficient for direct wastewater testing.
Does a novel multiplex PV2-specific assay improve detection of PV2 RNA in wastewater and clinical samples compared to standard isolation protocols?
Does a novel multiplex PV2-specific assay improve detection of PV2 RNA in wastewater and clinical samples compared to standard isolation protocols?
A novel PV2-specific RT-qPCR assay enables rapid, direct, and quantitative detection of poliovirus type 2 in clinical and environmental samples, improving surveillance capabilities.
BACKGROUND: An increase in the number of circulating vaccine-derived poliovirus (cVDPV) outbreaks has been reported worldwide in recent years, necessitating improved surveillance and diagnostic tools. The current standard protocol for polio detection is lengthy, labor-intensive, and inefficient for direct wastewater testing. METHODS: Here we describe a multiplex poliovirus 2 (PV2)-specific assay that directly detects PV2 RNA derived from Sabin-2 vaccine in wastewater and stool samples. The new PV2-specific assay was used for direct detection of PV2 in environmental and clinical samples. RESULTS: In silico analysis suggested that the assay is specific for PV2 originating from Sabin-2 vaccine and identifies 98.3% of available sequences, including some with <95% similarity to the vaccine reference sequence. We show that the assay is specific for PV2 and does not identify 19 other enterovirus strains, including Sabin-1 and Sabin-3 RNA. The new assay enabled fast detection of PV2 in suspected clinical samples, which was subsequently confirmed by isolation and sequencing. By analyzing 183 concentrated wastewater samples, we further show that the new assay specifically and directly identifies PV2 RNA, in the presence of PV1, PV3, and other enterovirus RNA. CONCLUSIONS: The assay provides quantitative results, allowing continuous monitoring of viral load dynamics. Comparison of the standard isolation protocol and the new assay showed excellent agreement in terms of specificity and sensitivity. This specific and rapid assay may therefore be a valuable tool for monitoring cVDPV2 outbreaks in real time, and promote global efforts for eradication and containment of poliovirus circulation.
No takes yet. Share an insight, caveat, or question.
Bar-Or et al. (2025) studied Poliovirus Type 2 (cVDPV2) outbreak (n=183). Multiplex poliovirus 2 (PV2)-specific real-time quantitative PCR assay vs. Standard isolation protocol was evaluated on Detection of PV2 originating from Sabin-2 vaccine. A novel multiplex PV2-specific real-time quantitative PCR assay identified 98.3% of available Sabin-2 sequences in silico and directly detected PV2 RNA in 183 wastewater samples.
Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context: