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Molecular Epidemiology of Non-Polio Enterovirus: Insights From L20B Cell Line Adaptation From Children With Acute

Muhammad Naveed Adil1,2,3, Muhammad Athar Abbas1, Muhammad Masroor Alam2

  • 1From the Department of Animal Genomics and Biotechnology, PARC Institute of Advanced Studies in Agriculture, National Agricultural Research Centre, Islamabad, Pakistan.

The Pediatric Infectious Disease Journal
|April 29, 2026
PubMed
Abstract

Insights

Non-polio enteroviruses (NPEVs) are a significant cause of acute flaccid paralysis (AFP) in Pakistan. Enhanced surveillance using VP1 sequencing is crucial for accurate diagnosis and public health strategies.

Area of Science:

  • Virology
  • Epidemiology
  • Public Health

Background:

  • Non-polio enteroviruses (NPEVs) are increasingly identified as causes of acute flaccid paralysis (AFP), mimicking poliomyelitis and hindering eradication efforts.
  • Limited molecular surveillance in Pakistan complicates the comprehensive characterization of NPEVs.
  • The L20B cell line, used for poliovirus detection, can support NPEV replication, leading to diagnostic challenges in AFP cases.

Purpose of the Study:

  • To characterize NPEVs in AFP cases in Pakistan.
  • To assess the utility of the L20B cell line and molecular methods for NPEV detection.
  • To inform public health strategies for AFP surveillance and control.

Main Methods:

  • Analysis of 4615 stool samples from AFP cases (children ≤15 years) between January 2021 and December 2022.
  • Identification of 435 NPEVs using L20B cell line and RT-PCR.
  • VP1 sequencing of 218 isolates, with 153 high-quality sequences obtained using an optimized primer set (224/222).
  • Phylogenetic analysis using MEGA X software.

Main Results:

  • NPEVs were frequently detected in L20B-positive AFP cases, indicating limited cell line specificity.
  • Enterovirus B, particularly Echovirus 7 and 11, predominated.
  • Geographic clustering was observed in Punjab, Khyber Pakhtunkhwa, and Sindh, with seasonal peaks in late summer/early autumn.
  • Phylogenetic analysis revealed localized Enterovirus B circulation with low genetic variation.

Conclusions:

  • Diverse NPEVs are relevant in post-polio surveillance, complicating AFP diagnosis.
  • Routine VP1 sequencing and optimized primers are essential for reducing diagnostic uncertainty.
  • Targeted seasonal and regional monitoring is vital for effective public health interventions.
Keywords:
L20B cell lineVP1 sequencingacute flaccid paralysisnon-polio enterovirusesphylogenetic analysis

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