Molecular detection and genotyping of HBV from HBsAg positive patients in a tertiary hospital in Nigeria.

BACKGROUND: Nigeria remains one of the countries with a high hepatitis B virus (HBV) burden in Africa. Reports have indicated the presence of mixed HBV genotypes in Nigeria; however, there is still paucity of data regarding mixed genotype infections particularly in the Southern part of the country. OBJECTIVE: Our aim is to determine the HBV genotype distribution among HBsAg-positive gastroenterology patients at the University College Hospital Ibadan, Nigeria. METHOD: Serum samples were screened for HBsAg by ELISA, and positive samples were genotyped by semi-nested multiplex PCR for HBV genotypes A, B, C, D, E and F. RESULTS: Data generated were analyzed in R-studio. A total of 81/90 (90%) of HBsAg-positive samples were successfully genotyped, and genotype A was most prevalent with 15.7%, while genotypes B and E were the least with 1.2% each. Genotypes A/C infection was the highest among mixed infections with 40% prevalence, while genotypes A/D were the least prevalent mixed infection with 4.8%. CONCLUSION: We advocate for a comprehensive genotype analysis in larger cohorts across Nigeria, to give a more comprehensive understanding of the distribution and prevalence of different HBV genotypes population wide.

VP7, VP4, and NSP4 genes of species a rotaviruses isolated from sewage in Nigeria, 2014/2015: partial sequence characterization and biophysical analysis of NSP4 (enterotoxin).

Species A rotavirus are an important cause of childhood gastroenteritis, and the main contributor to its pathogenicity is the enterotoxin (NSP4) protein. Some biophysical properties of partial NSP4 genes of RVAs isolated from sewage in Nigeria during 2014/2015 were investigated. Samples were typed by RT-PCR and Sanger sequencing of partial VP4, VP7 and NSP4 genes. Phylogeny identified lineages within genotypes, predicted glycosylation sites; hydrophobicity profiles and amino acid alignments were employed to determine some biophysical properties of the NSP4 protein. The VP7 sequences of our isolates were the most diversified, the majority of the isolates carried NSP4 genes of the E1 genotype. Genotype specific variations both in hydrophobicity and potential glycosylation were identified, mutations were highest within the H3 hydrophobic domain and VP4 binding domain. The study of RVA NSP4 genes from non-clinical samples revealed that there were structural consistencies with those of reference genes.

Dynamics of CD4 cell count among HIV-infected individuals in Lagos, Nigeria.

Human immunodeficiency virus (HIV) infection leads to progressive loss of CD4 T cells. Antiretroviral therapy has been able to inhibit this process, resulting in significant level of immune recovery and function. Our aim is to investigate the dynamics of CD4 recovery among HIV patients in Lagos, Nigeria. A total of 213 HIV-positive individuals were enrolled between October 2007 and May 2008, and followed up for 9 months based on CD4 count. CD4 analysis was done by flow cytometry at enrollment and after every 3 months. Data were grouped according to age range, antiretroviral treatment (ART), and time between infection and diagnosis. Kaplan-Meier survival analysis was used for data analysis. There was a significant difference in CD4 count between antiretroviral (ART) naïve and ART experienced subjects (P < 0.001). About 50% of the ART experienced population was identified to show poor CD4 reconstitution unable to achieve a CD4 of 500 cells/µl after 9 months of therapy. Time interval between infection and therapy was also identified to contribute to poor CD4 restoration. Further studies need to be done to classify immunological nonresponders among HIV patients in Nigeria. We also recommend introduction of programs that will facilitate early detection of HIV infection.

CD4 Decay Rate as an Indicator of the Time Interval between Initial Infection and First Diagnosis among Drug-Naïve Human Immunodeficiency Virus Seropositive Individuals in Lagos, Nigeria.

OBJECTIVE: The aim of this study was to determine the time interval between human immunodeficiency virus (HIV) infection and the first diagnosis among drug-naïve individuals in Badagry, Nigeria. SUBJECTS AND METHODS: A sample of 213 subjects who tested HIV positive for the first time were enrolled in this study. The HIV diagnosis was performed using Startpak® and Determine® kits, and a CD4 count was carried out using a FACS Count® flow cytometer. The mean CD4 values were determined by gender and age groups. The time interval between initial HIV infection and first testing was calculated based on the average CD4 decay rate per calendar year, and data analysis was performed using SPSS software. RESULTS: At diagnosis, the mean CD4 values showed that females recorded 270 cells/µl and males 244 cells/µl. By age range, individuals <25 years recorded 437 cells/µl, those between 25 and 40 years of age had 237 cells/µl, and those aged ≥41 years had 192 cells/µl. There was a significant difference between CD4 cell categorization and age range (p < 0.001). Subjects aged between 25 and 40 years recorded the highest distribution of all CD4 cell counts. The time interval between infection and testing for females was 8.1 years and for males 6.7 years. Within the age group <25 years the interval was 5.1 years, whilst it was 8.1 years for those aged ≥41 years. CONCLUSION: Most of the population presented for testing during the advanced stages of infection. We suggest an upscaling of HIV voluntary counseling and testing to encourage early detection and better treatment outcomes.

Seroprevalence rates of hepatitis C virus (HCV) antibody and hepatitis B virus surface antigen (HBsAG) in blood donors in a Southwestern Nigerian city.

Hepatitis B and C have been identified as major causes of Transfusion transmitted infections, in Nigeria. Our objective was to determine the prevalence of Hepatitis B virus (HBV) and Hepatitis C virus (HCV) in prospective blood donors in Abeokuta, Nigeria. 305 blood donors were screened for the presence of Hepatitis B virus surface antigen (HBsAg) and HCV using a rapid immunochromatographic kit (DiaSpot®). Demographic information was also collected. Males constituted 96.4%, singles were the majority with 65%. Prevalence of HBsAg was 9.8%, HCV 1.3%, and dual positivity 0.3%. Prevalence of HBsAg and HCV among males was 10.2% and 1.4%, while females recorded 0.0% for HCV and HBsAg. Dual positivity was recorded in a male (0.33%). Analysis of the study variables revealed that only educational status was statistically associated with positivity of HBsAg [Formula: see text], HCV prevalence was highest in the illiterate group it was not statistically significant (P > 0.05). We report the prevalence rates of anti-HCV Ab and HBsAg in blood donors from Ogun State, Nigeria. Our results reveals higher rate of HBsAg and evidence of co-infection with both viruses, illiteracy was the only variable associated with HBV infection. We advocate for the inclusion of anti-HBc or HBeAg in donor screening in our environment.

Prevalence of HIV, HBV and HCV among livestock merchants and slaughterhouse workers in Ibadan, Nigeria.

Background: Most studies on viral infections among livestock handlers have focused on occupational exposure from inadvertent contact with infected animals. Consequently, little emphasis is given to the effect of their lifestyle on the acquisition of other blood-borne viruses. Objectives: To determine the prevalence and assess risk factors for HIV, HBV and HCV infections among livestock handlers in Ibadan, Nigeria. Methods: Blood samples were collected from 265 livestock handlers between October 2016 to April 2017 in Ibadan. The samples were tested for the presence of antibodies to HIV and HCV; and surface antigen to HBV using ELISA. Structured questionnaire was administered to collect information on risk factors associated with the transmission of these viruses. Data analysis was carried out using Chi-square test and logistic regression to determine the association between risk factors and predictors of infection (p < 0.05). Results: Of 265 participants, 11 (4.2%), 29 (10.9%) and 13 (4.9%) individuals tested positive for HIV, HBV and HCV infections respectively. Two (0.8%) of the participants were coinfected with HIV and HBV while 1(0.4%) was coinfected with both HBV and HCV. Individuals who travelled frequently in the course of Livestock trades had a higher rate of HIV infection. Conclusions: A high Infection with HIV, HBV and HCV is common among the study participants. There is a need for continued surveillance and awareness creation on preventive measures against these viruses.

Prevalence of hepatitis B virus core antibodies among blood donors in Nigeria: Implications for blood safety.

Background: Anti-hepatitis B core antibody (anti-HBc) testing improves transfusion safety by detecting past and current hepatitis B virus (HBV) infection while detecting hepatitis B surface antigen (HBsAg) in serology-negative HBV infection. However, occult HBV infection (OBI) (serum or liver HBV DNA-positive but HBsAg-negative) remains unaddressed among replacement blood donors - family members or friends who donate to replace blood transfused to a relative. Objective: This study assessed risk factors for a positive anti-HBc test among donors with OBI and determined the anti-HBc-positive status of replacement donors. Methods: The study was conducted at the University College Hospital Blood Bank, Ibadan, Nigeria, using blood samples collected from blood donors between April 2019 and May 2019. Donors were screened for HBsAg by rapid diagnostic test (RDT) and enzyme-linked immunosorbent assay (ELISA) and anti-HBc by ELISA, while HBV DNA was detected using a semi-nested polymerase chain reaction. Results: Of the 274 participants, 15 (5.5%) were HBsAg-positive by RDT and 36 (13.1%) by ELISA, while 133 (48.5%) were anti-HBc positive. Out of 232 HBsAg-negative donors, 107 (46.1%) were anti-HBc positive. Of the 107 HBsAg-negative but anti-HBc-positive samples, only one (0.93%) was HBV DNA-positive. The HBV DNA-positive donor was HBsAg-negative by both RDT and ELISA tests. Conclusion: This study establishes a potential risk for HBV transmission from isolated anti-HBc-positive donors to blood recipients. HBc immunoglobulin (antibody) M testing to identify blood units requiring further screening with polymerase chain reaction to detect OBI can prevent HBV transmission through blood transfusion.

Oral human papilloma virus infection among dental clinic attendees in Ibadan, Nigeria.

Background: Human papilloma virus (HPV) is associated with a subset of oropharyngeal squamous cell carcinoma and mouth or throat warts. However, there is currently limited information about oral HPV infections in Nigeria. Objective: This study aimed to provide information on the occurrence and circulating genotypes of HPV among patients attending three (one government and two private) dental clinics in Ibadan, Nigeria. Methods: An oral swab was collected from 231 dental clinic attendees in Ibadan between January 2016 and March 2017 and tested for HPV DNA by polymerase chain reaction targeting the E6/7 genes of the virus. Results: Twenty-three of the 231 swab samples were HPV DNA positive comprising 16 mono-infections and seven co-infections in 13 males and ten females. Genotype 16 was present in ten patients, genotype 6/11 in five, Genotype 18 and genotype 33 in four each, genotype 31 in three and genotype 39 in one. Twenty-one cases were high-risk HPV genotypes, while two were low-risk. Samples had co-infection and five had low risk type 6/11 either as single or as co-infection. Persons who had engaged in oral sex as well as those aged 21-30 years has significantly higher prevalence. Conclusion: This study showed that although HPV genotype 16 is the most common type among dental clinic attendees in Ibadan, other genotypes are also circulating and that oral sex is a risk factor for the infection. Therefore, introducing a multivalent HPV vaccine will reduce the risk of HPV-associated oropharyngeal carcinoma and other cancers in Nigeria.

Assessment of hepatitis B surface antigen negative blood units for HBV DNA among replacement blood donors in a hospital based blood bank in Nigeria.

BACKGROUND: Hepatitis B virus infection is one of the greatest threats to blood safety all over the world. The laboratory algorithm based on only the detection of hepatitis B surface antigen (HBsAg) leaves a gap for infected HBsAg negative donors to donate blood during the "window period" (WP) and late stages of infection. OBJECTIVE: To estimate the frequency of the presence of HBV deoxyribonucleic acid (DNA) in HBsAg negative blood units screened using two different assays for HBsAg in a high endemic region. METHODS: Frozen serum aliquot of 100 replacement blood donors who donated blood units that were HBsAg negative were retrieved and tested for HBV DNA. Sample positive for HBV DNA was sequenced by Sanger's method, genotyped and the viral load was determined. RESULTS: One sample (1%) was positive for HBV DNA. The HBV viral load of the sample was 768,000 IU/ml. The partial S-gene of the Hepatitis B virus isolated was genotype E using the NCBI viral genotyping tool. CONCLUSIONS: There is still a risk of HBV infected blood unit escaping detection when donor testing is limited to HBsAg screening. The use of NAT which can substantially reduce HBV infected blood donors from the donor pool should be considered.

Profiles of mutations in hepatitis B virus surface and polymerase genes isolated from treatment-naïve Nigerians infected with genotype E.

Introduction. Hepatitis B virus (HBV) infection is the leading cause of hepatitis, liver cirrhosis, and hepatocellular carcinoma (HCC). HBV genotype E (HBV/E) is the predominant genotype in West Africa and has been linked epidemiologically with chronic and occult HBV infections as well as development of HCC. Mutations in the surface and polymerase genes of HBV have been associated with occult infection, drug resistance, vaccine escape, as well as HCC.Hypothesis/Gap Statement. There is limited data on the occurrence and patterns of mutations associated with occult infection, drug resistance, vaccine escape and HCC for HBV/E.Aim. This study characterized amino acid (aa) substitutions in the major hydrophilic (MHR) and reverse transcriptase (RT) regions of the surface and polymerase genes respectively of HBV sequences from a group of Nigerians with genotype E infection. The CpG islands of the PreC/C and PreS/S regions of these sequences were also described.Methodology. HBV surface and polymerase genes were detected using PCR techniques. Occurrence of new and previously described mutations in these genes were analysed using phylogenetic techniques.Results. Overall 13 HBV isolates were each sequenced for polymerase and surface genes mutations. Thirteen and nine PreS/S and PreC/C HBV genes respectively were analysed for CpG islands. Mutations in the MHR and a-determinants region of the S protein were discovered in eleven and nine of the 13 tested isolates respectively. These mutations were concomitant with aa changes in the RT functional domains of the isolates. Mutations associated with vaccine escape, occult infection and poor HCC prognosis were identified in HBV/E isolated in this study. Furthermore, all the isolates had at least one putative nucleotide analogue resistance mutations. Drug resistance mutations had the highest association with CpG islands.Conclusion. The results of this study contribute to further understanding of HBV variability in Nigeria and the West African region. This will aid the planning of adequate HBV immunization and treatment programmes for the countries in the region.

Phylogenetic analysis of human parvovirus b19 sequences from eleven different countries confirms the predominance of genotype 1 and suggests the spread of genotype 3b.

Phylogenetic analysis of 166 human parvovirus B19 sequences from 11 different countries attributed 91.57% to genotype 1, 5.42% to genotype 3b, and 3.01% to genotype 3a. Very similar viruses of genotype 1 circulated widely in Europe and Israel. Genotype 3b seems to show an increasing spread outside of Africa.

Virus genes and host correlates of pathology are markedly reduced during respiratory syncytial and influenza virus co-infection in BALB/c mice.

Globally, influenza A virus (IAV) and respiratory syncytial virus (RSV) infection remain very high. There is also a high burden of IAV and RSV co-infection in developing countries. To develop universally protective vaccines against these infections, it is imperative that viral genes and immune correlates of pathology are elucidated. As such, we profiled virus genes expressions, histopathology and immunological responses of BALB/c mice infected with RSV and/or IAV in this study. RSV A2 and/or influenza A/H3N2/Perth/16/09 (Pr/H3N2) were induced over a seven-day period in BALB/c mice. Anaesthetized BALB/c mice (12-14 g) were divided into six groups (15-20 mice per group), inoculated with 32 µl each of 3LD50 Pr/H3N2 and/or 100 TCID50 RSV. Two groups (R or I) received RSV or Pr/H3N2 intranasally. Prior infection with either RSV or Pr/H3N2 was followed with a second challenge of the other virus 24 hours post inoculation in RI and IR groups. Another set was exposed to the two viruses simultaneously (I + R group) while the last group served as healthy controls. Five to seven mice per group were euthanized at days 2, 4 and 7. Lung and spleen organs were harvested for virus genes quantitation and immune cells phenotyping respectively. I + R group showed progressive downregulation of RSV F, G, NS1 and NS2 genes. IAV PB2 and M genes had high fold increase on day 2 and 4 post infections. However, by day 7 post infection, M and PB2 fold increase was lower. Also, increased proportions of NKT and T cell subsets were observed throughout the period in I + R group. Conversely, I group was characterized by reduced NKT cell counts and enhanced CD8 T cells levels while R group only showed an increased proportion of CD8 T cells towards the peak of infection. This study shows that RSV and IAV co-infection lead to reduced virulence and pathology compared to single infections. This information is very useful in combinatorial RSV/IAV vaccine design and development.

Measles Virus Infection Among Vaccinated and Unvaccinated Children in Nigeria.

This study investigated measles infection in vaccinated and unvaccinated children presenting with fever and maculopapular rash during measles outbreaks in the southern and western states of Nigeria. Measles, an acute viral illness caused by a virus in the family Paramyxoviridae, is a vaccine-preventable disease. Measles outbreak is common in Nigeria, despite the national immunization program. Children presenting with symptoms of measles infection in general hospitals and health centers in the states of southern and western Nigeria were recruited for this study. Vaccination history, clinical details, and 5 mL of blood were obtained from the children. Their sera samples were screened for specific immunoglobulin M antibodies to measles virus. Of 234 children tested (124 [53.2%] female), 133 (56.8%) had previously been vaccinated against measles virus, while 93 (39.7%) had not been vaccinated. Vaccination information for eight children could not be retrieved. One hundred and forty-three (62.4%) had measles IgM antibodies. Of these, 79 (55.3%) had been vaccinated for measles, while 65 (44.7%) had not. Despite the ongoing vaccination program in Nigeria, a high number of children are still being infected with measles, despite their vaccination status. Therefore, there is need to identify the reason for the low level of vaccine protection.