Key result
Direct detection methods targeting the VP4-VP2 region showed higher sensitivity (limit of detection = 25 copies) than VP1-targeted methods and successfully detected poliovirus in all samples.
Why the study?
Under current biorisk regulations, polio surveillance using virus isolation faces challenges in reporting timeliness, test sensitivity, and containment of poliovirus isolates.
Direct detection methods targeting the VP4-VP2 region show high sensitivity for poliovirus detection in stool samples, offering a potential alternative to current surveillance algorithms.
VP4-VP2 DD may speed PV detection in AFP surveillance; leaves open prospective validation before replacing culture methods.
Polio surveillance in the Global Polio Eradication Initiative has been conducted with virus isolation from stool samples of acute flaccid paralysis (AFP) cases. Under the current biorisk management/regulations, challenges arise in the timelines of the report, sensitivity of the test and containment of poliovirus (PV) isolates. In the present study, we evaluated protocols of previously reported direct detection (DD) methods targeting the VP1 or VP4-VP2 regions of the PV genome in terms of sensitivity and sequencability. An optimized protocol targeting the entire-capsid region for the VP1 sequencing showed a high sensitivity (limit of detection = 82 copies of PV genome) with a simpler and faster reaction than reported ones (i.e., with the addition of all the primers at the start of the reaction, the RT-PCR reaction finishes within 2.5 h). The DD methods targeting the VP1 region detected PV in 60 to 80% of PV-positive stool samples from AFP cases; however, minor populations of PV strains in the samples with virus mixtures were missed by the methods. Sequencability of the DD methods was primarily determined by the efficiency of the PCRs for both Sanger and nanopore sequencing. The DD method targeting the VP4-VP2 region showed higher sensitivity than that targeting the VP1 region (limit of detection = 25 copies of PV genome) and successfully detected PV from all the stool samples examined. These results suggest that DD methods are effective for the detection of PV and that further improvement of the sensitivity is essential to serve as an alternative to the current polio surveillance algorithm.
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Ueno et al. (2023) studied Acute flaccid paralysis (AFP) / Poliovirus. Direct detection methods targeting VP1 or VP4-VP2 regions vs. Current virus isolation methods was evaluated on Sensitivity and sequencability for poliovirus detection. Direct detection methods targeting the VP4-VP2 region showed higher sensitivity (limit of detection = 25 copies) than VP1-targeted methods and successfully detected poliovirus in all samples.
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