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Updated: Jul 21, 2026
Particle Agglutination Method for Poliovirus Identification
Published on: April 20, 2011
Direct detection of polioviruses using a recombinant poliovirus receptor
Nancy Gerloff1, Mark Mandelbaum1, Hong Pang2
1Division of Viral Diseases, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, United States of America.
Abstract:
Polioviruses are positive-sense, single-stranded RNA picornaviruses and the principal cause of poliomyelitis. Global poliovirus surveillance has relied on poliovirus isolation in cells, which may take a minimum of 10 days, involves maintaining two cell lines, and propagates virus in high titers. With eradication underway, a major objective of the Global Polio Eradication Initiative (GPEI) is to develop culture-independent detection of polioviruses as an alternative method to complement the current virus isolation technique. A culture-independent method on poliovirus-positive stool suspensions was assessed with commercially available recombinant soluble poliovirus receptor (PVR) coupled to Histidine (His) tags. Viral RNA was screened by quantitative real-time reverse transcription PCR using the poliovirus intratypic differentiation kit. Poliovirus recovery was optimized with PVR-His-tagged protein and buffers supplemented with polyethylene glycol. To validate the poliovirus-PVR-His tag purification assay, 182 poliovirus-positive stools of programmatic importance were parallel tested against the GPLN-accepted virus isolation method. The PVR-His tag enrichment method detected poliovirus in 164 of 171 poliovirus-positive stools, whereas the virus isolation method misidentified 38 stools as poliovirus-negative (McNemar χ2 p<0.0001). Using this method in combination with RNA extraction, viral RNA recovery increased and showed similar (WPV1) or higher (Sabin 1) sensitivity than the World Health Organization accredited variation of the virus isolation method. The PVR-His enrichment method could be a viable addition to poliovirus surveillance; similar methods have the potential to capture other human pathogens such as EV71 using an appropriate soluble His tag receptor.
Insights
A new culture-independent method using poliovirus receptor (PVR)-Histidine (His) tags enhances poliovirus detection in stool samples. This PVR-His tag enrichment offers a faster, more sensitive alternative to traditional virus isolation for polio surveillance.
Area of Science:
- Virology
- Molecular Biology
- Public Health
Background:
- Polioviruses cause poliomyelitis and global surveillance relies on cell-based virus isolation, a method that is time-consuming (≥10 days) and resource-intensive.
- The Global Polio Eradication Initiative (GPEI) seeks culture-independent methods to complement existing virus isolation techniques for enhanced poliovirus surveillance.
Purpose of the Study:
- To assess a novel culture-independent method for poliovirus detection using recombinant soluble poliovirus receptor (PVR) coupled to Histidine (His) tags.
- To optimize poliovirus recovery using the PVR-His tag enrichment method and evaluate its performance against the established virus isolation technique.
Main Methods:
- A PVR-His tag enrichment assay was developed and optimized using poliovirus-positive stool suspensions, polyethylene glycol, and specific buffers.
- Viral RNA was screened using quantitative real-time reverse transcription PCR (qRT-PCR) with a poliovirus intratypic differentiation kit.
- The PVR-His tag purification assay was validated by parallel testing of 182 poliovirus-positive stools against the Global Polio Laboratory Network (GPLN)-accepted virus isolation method.
Main Results:
- The PVR-His tag enrichment method detected poliovirus in 164 of 171 (95.9%) positive stools, significantly outperforming virus isolation, which misidentified 38 stools as negative (McNemar χ2 p<0.0001).
- Combined with RNA extraction, the PVR-His method demonstrated comparable (WPV1) or superior (Sabin 1) sensitivity to the World Health Organization (WHO)-accredited virus isolation method.
- The method showed potential for detecting other human pathogens, such as Enterovirus 71 (EV71), using appropriate His tag receptors.
Conclusions:
- The PVR-His tag enrichment method represents a viable, culture-independent addition to poliovirus surveillance strategies.
- This approach offers improved sensitivity and efficiency over traditional cell-based isolation, crucial for ongoing eradication efforts.
- Similar His tag receptor-based enrichment methods hold promise for the detection of a broader range of viral pathogens.
