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Nonpolio enteroviruses can also be recovered from non-reproducible cytopathology in L20B cell line

View ORCID ProfileJ. A. Adeniji, U.I. Ibok, O.T. Ayinde, A.O. Oragwa, U.E. George, View ORCID ProfileT.O.C. Faleye, View ORCID ProfileM.O. Adewumi
doi: https://doi.org/10.1101/158626
J. A. Adeniji
1Department of Virology, College of Medicine, University of Ibadan, Ibadan, Oyo State, Nigeria
2WHO National Polio Laboratory, University of Ibadan, Ibadan, Oyo State, Nigeria
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U.I. Ibok
1Department of Virology, College of Medicine, University of Ibadan, Ibadan, Oyo State, Nigeria
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O.T. Ayinde
2WHO National Polio Laboratory, University of Ibadan, Ibadan, Oyo State, Nigeria
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A.O. Oragwa
1Department of Virology, College of Medicine, University of Ibadan, Ibadan, Oyo State, Nigeria
3Department of Veterinary Microbiology and Pathology, Faculty of Veterinary Medicine, University of Jos, Jos, Plateau State, Nigeria
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U.E. George
1Department of Virology, College of Medicine, University of Ibadan, Ibadan, Oyo State, Nigeria
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T.O.C. Faleye
1Department of Virology, College of Medicine, University of Ibadan, Ibadan, Oyo State, Nigeria
4Department of Microbiology, Faculty of Science, Ekiti State University, Ado-Ekiti, Ekiti, State, Nigeria
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M.O. Adewumi
1Department of Virology, College of Medicine, University of Ibadan, Ibadan, Oyo State, Nigeria
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ABSTRACT

Samples showing cytopathology (CPE) on initial inoculation into L20B cell line but with no observed or reproducible CPE on passage in L20B or RD are considered negative for both poliovirus and nonpolio enteroviruses (NPEVs). The phenomenon is termed ‘non-reproducible CPE’. Its occurrence is usually ascribed to the likely presence of reoviruses, adenoviruses and other non-enteroviruses. This study aimed to investigate the likelihood that NPEVs are also present in cases with non-reproducible CPE.

Twenty-six (26) cell culture suspensions were analyzed in this study. The suspensions were collected from the WHO National Polio Laboratory, Department of Virology, College of Medicine, University of Ibadan. The suspensions emanated from 13 L20B cell culture tubes that showed cytopathology within 5 days of inoculation with fecal suspension from AFP cases. However, on passage into one each of RD and L20B cell lines, the CPE was not reproducible. All samples were subjected to RNA extraction, cDNA synthesis, the WHO recommended VP1 RT-seminestedPCR assay, species resolution PCR assay, sequencing and phylogenetic analysis.

Six (6) samples were positive for the VP1 RT-seminested PCR assay. Only four of which were positive by the species resolution PCR assay. The four amplicons were sequenced, however, only three (3) were successfully identified as Coxsackievirus A20 (2 isolates) and Echovirus 29 (1 isolate).

The results of this study unambiguously showed the presence of NPEVs (particularly CVA20 and E29) in cell culture supernatants of samples with CPE on initial inoculation into L20B cell line but with no observed or reproducible CPE on passage in RD cell line. Therefore, like reoviruses, adenoviruses and other non-enteroviruses, NPEVs can also be recovered in cases with non-reproducible CPE.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted July 02, 2017.
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Nonpolio enteroviruses can also be recovered from non-reproducible cytopathology in L20B cell line
J. A. Adeniji, U.I. Ibok, O.T. Ayinde, A.O. Oragwa, U.E. George, T.O.C. Faleye, M.O. Adewumi
bioRxiv 158626; doi: https://doi.org/10.1101/158626
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Nonpolio enteroviruses can also be recovered from non-reproducible cytopathology in L20B cell line
J. A. Adeniji, U.I. Ibok, O.T. Ayinde, A.O. Oragwa, U.E. George, T.O.C. Faleye, M.O. Adewumi
bioRxiv 158626; doi: https://doi.org/10.1101/158626

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