Abundance of enterovirus C in RD-L20B cell culture-negative stool samples from acute flaccid paralysis cases in Nigeria is geographically defined

J Med Microbiol. 2018 Jun;67(6):854-865. doi: 10.1099/jmm.0.000737. Epub 2018 Apr 30.

Abstract

Purpose: We recently showed that enteroviruses (EVs) andenterovirus species C (EV-C) in particular were abundant in faecal samples from children who had been diagnosed with acute flaccid paralysis (AFP) in Nigeria but declared to be EV-free by the RD-L20B cell culture-based algorithm. In this study, we investigated whether this observed preponderance of EVs (and EV-Cs) in such samples varies by geographical region.

Methodology: One hundred and eight samples (i.e. 54 paired stool suspensions from 54 AFP cases) that had previously been confirmed to be negative for EVs by the WHO-recommended RD-L20B cell culture-based algorithm were analysed. The 108 samples were made into 54 pools (27 each from North-West and South-South Nigeria). All were subjected to RNA extraction, cDNA synthesis and the WHO-recommended semi-nested PCR assay and its modifications. All of the amplicons were sequenced, and the enteroviruses identified, using the enterovirus genotyping tool and phylogenetic analysis.

Results: EVs were detected in 16 (29.63 %) of the 54 samples that were screened and successfully identified in 14 (25.93 %). Of these, 10 were from North-West and 4 were from South-South Nigeria. One (7.14 %), 2 (14.29 %) and 11 (78.57 %) of the strains detected were EV-A, EV-B and EV-C, respectively. The 10 strains from North-West Nigeria included 7 EV types, namely CV-A10, E29, CV-A13, CV-A17, CV-A19, CV-A24 and EV-C99. The four EV types recovered from South-South Nigeria were E31, CV-A1, EV-C99 and EV-C116.

Conclusion: The results of this study showed that the presence of EVs and consequently EV-Cs in AFP samples declared to be EV-free by the RD-L20B cell culture-based algorithm varies by geographical region in Nigeria.

Keywords: Enteroviruses; Nigeria; RD-L20B; acute flaccid paralysis; cell culture negative samples.

MeSH terms

  • Acute Disease
  • Adolescent
  • Bacteriological Techniques
  • Cell Line
  • Child
  • Child, Preschool
  • Enterovirus C, Human / classification
  • Enterovirus C, Human / genetics*
  • Enterovirus C, Human / growth & development
  • Enterovirus C, Human / isolation & purification*
  • Enterovirus Infections / epidemiology*
  • Enterovirus Infections / virology
  • Feces / virology*
  • Female
  • Humans
  • Male
  • Nigeria / epidemiology
  • Paraplegia / virology*
  • Phylogeny
  • Polymerase Chain Reaction