Virology
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Item Isolation and Identification of Enteroviruses from Sewage and Sewage-Contaminated Water Samples from Ibadan, Nigeria, 2012-2013(SciTechnol, 2017) Adeniji, J. A.; Adewale, A. O.; Faleye, T. O. C.; Adewumi, M. O.In 2010, we described sewage contaminated water (SCW) bodies that consistently yielded enteroviruses (EVs) in enterovirus surveillance (ES) sites in Lagos, Nigeria. By 2012, we demonstrated the presence and circulation of Wild Poliovirus 3 (WPV3) in these ES sites. Here we describe ES sites that consistently yield EVs in Ibadan metropolis southwest Nigeria. Twenty-five ES samples were collected by grab method from nine sites between October, 2012 and March, 2013. Samples were concentrated and four (RD, HEp2C, MCF-7 and L20B) different cell lines used for virus isolation from the concentrates. Isolates were subjected to RNA extraction, cDNA synthesis, PanEnterovirus 5l-UTR and VP1 assays. Unidentifiable isolates were further subjected to species-specific RTPCR assays. Amplicons were sequenced, isolates identified and subjected to phylogenetic analysis. Twenty-five isolates were recovered from 8 (32%) of the samples collected. Twenty-three of the isolates were identified as EVs by the PanEntero5l-UTR assay. Thirteen (57%) of the 23 EVs were positive for the VP1 assay, and identified as Coxsackievirus B3 (CVB3) (1 isolate), CVB6 (1 isolate), E6 (2 isolates), E7 (5 isolates), E11 (1 isolate), E12 (1 isolate) and E13 (2 isolates). None and 2 (25%) of the remaining isolates were positive for the EV-B and EV-C assays, respectively. The 2 EV-C positive enteroviruses were isolated on MCF-7. This study describes three very productive ES sites, and documents the presence of CVB3, CVB6, E6, E7, E11, E12 and, E13 in Ibadan, Nigeria. Including other cell lines in EV isolation protocols can broaden the diversity of EV types recoverable.Item The Impact of a Panenterovirus Vp1 Assay on Our Perception of the Enterovirus Diversity Landscape of a Sample(MedCrave Group, 2016) Faleye, T. O. C.; Adewumi, M. O.; Kareem, S. A.; Adesuyan, Y. O. Fapohunda, F. A.; Fasanya, S. T.; Jimeto, T.; Lawrence, O .E.; Obembe, A. A.; Adeniji, J. A.The number of independent emergence of recombinant circulating vaccine derived poliovirus serotype 2 (cVDPV2) lineages and the magnitude of the outbreak in Nigeria demonstrates the significance of enterovirus co-infection and its preponderance in the country. Despite this, besides polioviruses, little or no attention is given to enterovirus co-infections. More recently, a reverse-transcriptase semi-nested polymerase chain reaction (RT-snPCR) assay for the direct detection of enteroviruses from clinical specimen was added to the WHO recommended protocols for enterovirus surveillance. We previously showed that primers 292 and 222 (for which AN89 and AN88, respectively, are consensus degenerate hybrid oligonucleotide primers [CODEHOP] versions) mask the presence of enterovirus co-infections. We therefore ask whether the primers AN89 and AN88 used in this recently recommended RT-snPCR protocol, like primers 292 and 222, will also mask the presence of enterovirus co-infections. RNA was extracted from 30 archived samples (both clinical specimen and cell culture isolates) and the VP1 gene amplified using the WHO recommended RT-snPCR assay and modifications that included the primers 187, 188 and 189 for which primer 292 (and consequently AN89) is a consensus. Amplicons were sequenced and isolates identified. Our results showed that primers AN89 and AN88 also mask enterovirus co-infection and inclusion of the primers 187, 188 and 189 allowed the resolution of such mixed isolates. Consequently, expanding the recommended RT-snPCR protocol to include primers 187, 188 and 189 will enable us better detect enterovirus co-infectionItem Enterovirus A119 in A Child with Acute Flaccid Paralysis, Nigeria(MedCrave Group, 2016) Adeniji, J. A.; Oragwa, A. O.; George, U. E.; Ibok, U. I.; Faleye, T. O. C.; Adewumi, M. O.The oldest EV-A119 record was in 2008 in a chimpanzee in Cameroon and subsequently in more non-human primates and healthy children. Here we report for the first time the detection of EV-A119 in a child with Acute Flaccid Paralysis, thus suggesting possible association with a clinical condition in humans