Virology
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Item Isolation and Complete Capsid Sequence of Enterovirus D111 from Faeces of a Child with Acute Flaccid Paralysis in Nigeria(ResearchersLinks Ltd, 2019) Faleye, T. O. C.; Adewumi, M. O.; Olayinka, O. T.; Adeniji, J. A.The WHO recommended cell-culture-based algorithm requires enterovirus (EV) isolates to produce reproducible cytopathic effect (R-CPE) in RD and/or L20b cell lines. Samples with non-reproducible CPE (NR-CPE) are considered negative for EVs. We investigated whether there could be EVs lurking in samples with NR-CPE. Fifty-nine cell culture supernatants (CCS) (collected between 2016 and 2017) from RD and L20b cell-culture-tubes with NR-CPE, were analyzed. The tubes had been previously inoculated with stool suspension from children (<15 years) in Nigeria with acute flaccid paralysis (AFP). All CCS were screened for Enteric Adenoviruses (EAVs) and group A Rotavirus (GARV) using a rapid immunochromatographic test kit. Subsequently, they were passaged in HEp-2 cell line. All isolates were subjected to RNA extraction, cDNA synthesis, three (5l-UTR, VP1 and EV Species C [EV-C]) different PCR assays and sequencing of amplicons. EVs were further subjected to [‘]Illumina sequencing. All CCS were negative for EAVs and GARVs. Four CCS produced R-CPE in HEp-2 cell line, three of which were positive for the 5l-UTR assay. Of the three isolates, two and none were positive for the VP1 and EV-C assays, respectively. One of the two VP1 amplicons was successfully sequenced and identified as Echovirus 1. Illumina sequencing of the three 5l-UTR positive isolates confirmed the E1 isolate and typed the remaining two as EV-Ds (94 and 111). We describe the first EV-D isolates of Nigerian origin and the first complete capsid sequence of the virus globally. We also show that NR-CPE could sometimes be caused by EVs that do not produce R-CPE in specific cell lines.Item Rapid HIV Antigen–Antibody Assays and detection of acute HIV infection in sub-saharan Africa(American Society of Tropical Medicine and Hygiene, 2019) Adetunji, A. A.; Adewumi, M. O.; Michael, O. S.; Fayemiwo, S. A.; Ogunniyi, A.; Taiwo, B. O.Detection of acute HIV infection is a unique problem that fourth-generation HIV assays were expected to alleviate. In this commentary, we draw attention to the limitations and challenges with use of currently available rapid antigen–antibody (Ag/Ab) combination tests for detection of acute HIV infection in sub-Saharan Africa. Laboratory-based HIV-1 Ag/Ab immunoassays are complex, requiring specialized equipment and handling that are currently not affordable in many settings in Africa. The point-of-care Ag/Ab platform on the other hand is easier to deploy and potentially more accessible in resource-limited settings. However, available fourth-generation HIV-1 rapid diagnostic tests have demonstrated poor performance characteristics in field studies where non-B subtypes of HIV-1 dominate. The potential for point-of-care HIV-1 Ag/Ab diagnostics to significantly improve detection of acute HIV infection remains yet to be realized in sub-Saharan Africa. Assay platforms need to be optimized to identify local circulating subtypes, and optimal algorithms need to be determinedItem Characterization of hepatitis delta virus strains spreading in Abuja, Nigeria(Wiley Periodicals, Inc.,, 2019) Ifeorah, I. M.; Faleye, T. O. C.; Bakarey, A. S.; Adewumi, O. M.; Gerber, A.; Le Gal, F.; Adeniji, J. A; Gordien, E.; Onyemelukwe, N. F.Hepatitis delta virus (HDV) is responsible for the most severe form of liver disease in humans. So far, eight genotypes (HDV‐1 to ‐8) have been individualized worldwide. Little is known about HDV strains that spread in Nigeria. HDV genotyping was performed in 15 anti–HDV positive samples from a cohort of 306 hepatitis B virus (HBV)‐infected patients in Abuja (Nigeria). Phylogenetic analyses revealed 90% were HDV‐1, two among them clustering with European/Asian HDV‐1, the remaining one being HDV‐6. It was also found that two members of a couple superinfected with the same HDV strain, were enveloped by two different HBV strains of genotype E.Item Towards an effective poliovirus laboratory containment strategy in Nigeria(BMC (BioMed Central),, 2018) Ticha, J. M.; Matthew, K. O.; Hamisu, A. W.; Braka, F.; Mkanda, P.; Nsubuga, P.; Tesfaye, E.; Craig, K,; Etsano, A.; Emelife, O.; Shuaib, F.; Akinkugbe, F.; Adeniji, J. A.; Adamu, U.; Dallatu, M.; Oyeyinka, G.; Brown, H.; Nnamah, N.; Okwori, J.; Chinedu, C.; Anibijuwon, I.; Adewumi, O. M.; Donbraye, E.; Bagana, M.; Baba, M.; Gumede, N.; Banda, R.; Tegegne, S. G.; Oyetunji, A.; Diop, O.; Tomori, O.; Vaz, R.G.Background: The Global Commission for the Certification of the Eradication of Poliomyelitis will declare the world free of wild poliovirus transmission when no wild virus has been found in at least 3 consecutive years, and all laboratories possessing wild poliovirus materials have adopted appropriate measures of containment. Nigeria has made progress towards poliomyelitis eradication with the latest reported WPV type 1 on 21 Aug 2016 after 2 years without any case. This milestone achievement was followed by an inventory of biomedical laboratories completed in November 2015 with the destruction of all identified infectious materials. This paper seeks to describe the poliovirus laboratory containment process in Nigeria on which an effective containment system has been built to minimize the risk of virus re-introduction into the population from the laboratories. Methods: A national survey of all biomedical facilities, as well as an inventory of laboratories from various sectors, was conducted from June November 2015. National Task Force (NTF) members and staff working on polio administered an on-site questionnaire in each facility. Laboratory personnel were sensitized with all un-needed materials destroyed by autoclaving and incineration. All stakeholders were also sensitized to continue the destruction of such materials as a requirement for phase one activities. Results: A total of 20,638 biomedical facilities were surveyed with 9575 having laboratories. Thirty laboratories were found to contain poliovirus or potentially infectious materials. The 30 laboratories belonged to the ministries of health, education, defence and private organizations. Conclusions: This article is amongst the first in Africa that relates poliovirus laboratory containment in the context of the tOPV-bOPV switch in alignment with the Global Action Plan III. All identified infectious materials were destroyed and personnel trained to continue to destroy subsequent materials, a process that needs meticulous monitoring to mitigate the risk of poliovirus re-introduction to the population.Item Draft genome sequence of a bovine enterovirus isolate recovered from sewage in Nigeria(American Society for Microbiology (ASM), 2018) Faleye, T. O. C; Adewumi, O. M.; Olayinka, O. A.; Donbraye, E; Oluremi, B.; George, U. E.; Arowolo, O. A.; Omoruyi, E. C.; Ifeorah, M. I.; Oyero, A . O.; Adeniji, J. A.We describe the draft genome of a bovine enterovirus (EV) isolate recovered from sewage in Nigeria. This isolate replicates on both RD and L20B cell lines but is negative for all EV screens in use by the Global Poliovirus Eradication Initiative (GPEI). It contains 7,368 nucleotides (nt) with 50.2% G C content and an open reading frame (ORF) with 6,525 nt (2,174 amino acids).Item Reference Echovirus 7 and 19 Genomes from Nigeria(American Society for Microbiology (ASM), 2018) Adeniji, J. A.; Faleye, T. O. C.; Adewumi, O. M.We describe the genomes of two echovirus isolates from Nigeria as reference enterovirus species B genomes for the region. These echovirus 7 and 19 genomes have 7,411 nucleotides (nt) and 7,426 nt and were recovered from sewagecontaminated water (in 2010) and an acute flaccid paralysis case (in 2014), respectively.Item Identification of previously untypable RD cell line isolates and detection of EV-A71 genotype C1 in a child with AFP in Nigeria(Taylor & Francis, 2018-11-26) Adewumi, O. M.; Faleye, T. O. C.; Okeowo, C. O.; Oladapo, A. M.; Oyathelemhi, J.; Olaniyi, O. A.; Isola, O. C.; Adeniji, J. A.We previously attempted to identify 96 nonpolio enteroviruses (EVs) recovered in RD cell culture from children <15years with acute flaccid paralysis in Nigeria. We succeeded in identifying 69 of the isolates. Here, we describe an attempt to identify the remaining 27 isolates. Twenty-six (the 27th isolate was exhausted) isolates/samples that could not be typed previously were further analyzed. All were subjected to RNA extraction, cDNA synthesis, enterovirus 5ʹ-UTR–VP2 PCR assay and a modified VP1 snPCR assay. Both the 5ʹ-UTR–VP2 and VP1 amplicons were sequenced, isolates identified and subjected to phylogenetic analysis. Twenty of the 26 samples analyzed were identified. Altogether, 23 (three samples had co-infection) EV strains were recovered. These belong to 11 EV (one EVA, nine EVB and one EVC) types which were EVA71 genotype C1 (1 strain), CVB3 (7 strains), CVB5 (1 strain), E5 (2 strain), E11 (3 strains), E13 (2 strain), E19 (1 strain), E20 (1 strain), E24 (2 strains), EVB75 (1strain) and EVC99 (2 strains). Of the 11 EV types, the 5ʹ-UTR-VP2 assay identified seven while the VP1 assay identified 10. Both assays simultaneously detected 7 of the 11 EV types identified in this study with 100% congruence. We successfully identified 20 of 26 samples that were previously untypable. We also provided evidence that suggests a clade of EVA71 genotype C1 might have been circulating in sub-Saharan Africa since 2008. Finally, we showed that the 5ʹ-UTR -VP2 assay might be as valuable as the VP1 assay in EV identification.Item Abundance of enterovirus C in RD-L20B cell culture-negative stool samples from acute flaccid paralysis cases in Nigeria is geographically defined(Microbiology Society, 2018) Donbraye, E.; Olasunkanmi, O. I.; Opabode, B. A.; Ishola, T. R.; Faleye, T. O. C; Adewumi, O. M.; Adeniji, J. A.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 thestrains 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.Item Draft Genome Sequence of Mycoplasma arginini Strain NGR_2017(Merican Society for Microbiology (ASM), 2018-06-28) Adeniji, J.A.; Faleye, T. O. C.; Adewu , M. O.; Olayinka O. A.; Donbraye, E.; Oluremi, B.; George,a U. E.; Arowolo, O. A.; Omoruy, E. C.; Ifeorah, M. I.; Akande, A.We present the draft genome of Mycoplasma arginini strain NGR_2017. This strain was recovered in Nigeria from cell culture in 2017. The assembly contains 620,555 bp in 12 contigs. It contains 561 coding sequences, 34 RNAs (29 tRNAs, 4rRNAs, and 1 transfer-messenger RNA [tmRNA]), and a_26-kb integrative and conjugative element.Item Evaluation of performance testing of different rapid diagnostic kits in comparison with EIAs to validate detection of hepatitis B virus among high risk group in Nigeria(Taylor & Francis, 2018-05-15) Afolabi, A. Y.; Bakarey, A. S.; Adewumi, M.O.Background: Hepatitis B virus (HBV) causes chronic liver-associated diseases and its early detection is of high public health importance. Its diagnosis is mainly based on immunological assays among which Enzyme-Linked Immunosorbent Assay (ELISA) and rapid tests are the most common and widespread methods. However, a major challenge is the discordance of results of any two laboratory assays which cannot be easily resolved. Therefore, this study was designed to evaluate the validity and reliability of commercially available five rapid test kits in comparison with two Enzyme Immunoassays (EIAs) in Nigeria using hepatitis B surface antigen as a reference marker. Methods: A total of 100 sera of previously diagnosed consenting HBV-positive patients from private diagnostic laboratories in Ibadan between March and August, 2011 were tested using two EIA and five rapid commercially available HBV test kits in Nigeria. Data were analyzed by SPSS version 15, while bivariate and multivariate analyses were carried out to identify associations at P < 0.05 considered significant. Results: Overall, the sensitivity rates of the two EIA kits were 100% and 99.9% (95% confidence interval [CI] = 98.9–99.7) with specificity of 100% and 99.9% (95% CI = 98.9–99.7), respectively. The sensitivity of the five rapid test kits ranged from 97.5% (95% CI = 96.4–97.6) to 98.9% (95% CI = 97.9–99.9) with specificity of 80% (95% CI = 79.3–80.9) to 90% (95% CI = 89.2–91.0). Also, the positive predictive value ranged from 88% (95% CI = 88.2–89.9) to 89% (95% CI = 88.2–89.9), while the negative predictive value ranged from 80% (95% CI = 79.3–80.9) to 90% (95% CI = 89.2–91.0) for the five rapid kits. However, that of the two EIAs ranged from 99.9% (98.9– 99.7) to 100%. Further analysis showed significant (P = 0.033) variations in the sensitivity and specificity of the EIAs and rapid test kits. Conclusions: The results from this study have clearly revealed the challenges of diagnosis of HBV infections in Nigeria. This study has also demonstrated that the sensitivity of most of the rapid test kits may not be adequate when compared with EIA for early detection of HBV infections. The implications of possible misdiagnosis on the various intervention strategies that rely predominantly on correct HBV status of an individual are enormous. Therefore, there is the need to further compliment the use of rapid test kits with EIAs for HBV control in Nigeria.