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Updated: Oct 23, 2026
Particle Agglutination Method for Poliovirus Identification
Published on: April 20, 2011
Poliovirus nonpermissive CD155 knockout cells derived from RD cell line for handling poliovirus potentially
Shyam S Nandi1, Sonali Sawant1, Trupti Gohil1
1National Institute of Virology Mumbai Unit, Haffkine Institute Compound, Mumbai, India.
Abstract:
Destruction of all poliovirus containing materials, safe and secure handling of retained polioviruses for vaccine production, and research will be obligatory to eliminate facility-associated risks. Polioviruses and poliovirus potentially infectious materials (PIM) including fecal or respiratory samples requiring containment have been defined in World Health Organization-Global Action Plan (GAP III) documents. Non-polio laboratories culturing viruses from PIM are most affected as cell cultures of human and monkey origin are also poliovirus permissive. CRISPR gene-editing technology was used to knockout the poliovirus receptor (PVR/CD155) gene in the rhabdomyosarcoma (RD) cell line. PVR knockout RD cell susceptibility was tested using known non-polio enterovirus (NPEV) types. A selected clone (RD-SJ40) was field evaluated for virus isolation from 626 stool samples of acute flaccid paralysis cases. Poliovirus nonpermissive cells derived from the RD cell line did not show CD155-specific cell-surface immunofluorescence. CD155 gene sequencing confirmed nucleotide base pair deletions within exon2 and exon3. The CD155 knockout RD-SJ40 cells did not support the growth of poliovirus from positive stool samples. All NPEV types were isolated in RD and RD-SJ40 cells. CRISPR correctly edited the CD155 gene of RD cells to render them poliovirus nonpermissive while susceptibility to NPEV remained unchanged. RD-SJ40 cells are safe for NPEV isolation from poliovirus PIM without derogating GAP III containment requirements.
Insights
Researchers created poliovirus non-susceptible cells using CRISPR gene editing. These RD-SJ40 cells safely isolate non-polio enteroviruses from infectious samples, aiding global polio eradication efforts.
Area of Science:
- Virology
- Molecular Biology
- Biotechnology
Background:
- Facility-associated risks necessitate safe handling of polioviruses and potentially infectious materials (PIM).
- Non-polio laboratories face challenges as standard cell cultures are permissive to poliovirus.
- World Health Organization's Global Action Plan (GAP III) outlines containment requirements for poliovirus PIM.
Purpose of the Study:
- To develop poliovirus non-susceptible cell lines for safe isolation of non-polio enteroviruses (NPEV).
- To evaluate the efficacy of CRISPR gene-editing technology in creating these modified cell lines.
- To ensure the modified cells retain susceptibility to NPEV for diagnostic purposes.
Main Methods:
- CRISPR gene-editing was employed to knockout the poliovirus receptor (PVR/CD155) gene in the rhabdomyosarcoma (RD) cell line.
- The PVR knockout RD cell line (RD-SJ40) was tested for susceptibility to various NPEV types.
- RD-SJ40 cells were field-evaluated for virus isolation from 626 stool samples from acute flaccid paralysis cases.
Main Results:
- CD155 gene sequencing confirmed successful nucleotide base pair deletions in the RD-SJ40 clone.
- RD-SJ40 cells demonstrated no poliovirus growth from positive stool samples.
- All tested NPEV types were successfully isolated in both RD and RD-SJ40 cells, indicating retained susceptibility.
Conclusions:
- CRISPR gene editing effectively rendered RD cells poliovirus non-permissive by knocking out the CD155 gene.
- The RD-SJ40 cell line is safe and suitable for NPEV isolation from poliovirus PIM.
- These modified cells support GAP III containment requirements while enhancing laboratory safety.
