This was a multicenter national hospital-based cross-sectional pilot baseline study designed to explore the HCV awareness, the prevalence of HCV and factors affecting such awareness and prevalence rates among pregnant women.
The study was conducted in Nigeria, a country made up of 36 states and the Federal capital territory grouped into six geopolitical zones. Participants were recruited from one randomly selected tertiary level health facility in each of the five geopolitical zones in Nigeria apart from the South East zone where the lead institution for the TETFund National Research Fund 2019 was located. The sites randomly selected are: Aminu Kano Teaching Hospital, Kano [North-West zone]; Obafemi Awolowo University Teaching Hospital Complex, Ile-Ife [South-West zone]; University of Port Harcourt Teaching Hospital, Port-Harcourt [South-South zone]; University of Abuja Teaching Hospital, Gwagwalada [North-Central zone]; University of Maiduguri Teaching Hospital, Maiduguri [North-East zone] in addition to the lead Institution, Nnamdi Azikiwe University Teaching Hospital, Nnewi [South-East zone]. A summary of the protocol is described here, and the complete protocol has previously been published .
The study was carried out among consenting pregnant women, who were attendees of the antenatal clinic in randomly selected health care facilities in each of the six geopolitical zones of Nigeria.
Pregnant women registered for their antenatal care in the study sites were eligible to participate in the study.
Non-pregnant women and women whose pregnancy could not be confirmed by ultrasound or blood test were excluded.
Recruitment of participants
We employed random sampling by selecting 6 tertiary hospitals across the 6 geopolitical zones in Nigeria. The research assistants were adequately trained in recruiting and screening pregnant women in the survey. Pregnant women were recruited from the antenatal clinics of each hospital over a one month (between June 2, 2020 and July 1, 2020) period. After detailed explanation of the objectives, procedures and possible benefits of the study, only those who accepted to participate in the study and gave a written informed consent were enrolled into the study. The participants were then interviewed using study specific tool. The study tool contained questions asking women whether they had heard of HCV infection, including whether they had specific risk factors for HCV such as past history of HBV, intravenous drug use, multiple sex partners, etc). Past history of HBV was defined as participants who previously tested positive to hepatitis B surface antigen. Intravenous drug use was defined as injection of chemicals into the body through a hypodermic needle into a vein. A person was defined to have multiple sex partners, when the person has sex with more than one person at the same time. Pregnant women who completed the survey, were asked for their contact details and given a sole identifier; a research team member cross checked these details to ensure each woman finished the survey questions. Thereafter, blood sample was collected for HCV determination.
Laboratory procedure and analysis
Five milliliters of the blood sample was aseptically collected by venipuncture from each pregnant woman into a plain specimen bottle after consent was sought and obtained. The participant’s code, age, time and date of collection were labeled on the bottle for proper identification. Each of the samples was centrifuged at 3,000/rpm for 5 min and the serum portion was used on the test strip for antigen or antibody detection. Sera samples were stored at -25oC in line with the kit manufacturer’s instruction until screened for HBsAg, anti-HIV, and anti-HCV antibodies. Screening for Anti-HCV antibodies was conducted using the Enzyme-linked immunosorbent assay (ELISA) kit manufactured by LabACON (Hangzhou Biotest Biotech Company, Ltd., China) which has a specificity of 99.0% and a sensitivity of 99.9% according to manufacturer declared figures. The kit has in-built controls. The manufacturer’s instruction was strictly followed and executed by trained research assistants in each facility. The results were reported as positive or negative. The Anti-HCV antibodies tests were confirmed using DNA PCR analysed centrally at Molecular Virology Laboratory, NAUTH, Nnewi, Nigeria. Serial rapid HIV testing was done according to Nigerian National HIV testing guidelines viz. Alere Determine HIV-1/2 (Alere Medical Co. Ltd., Matsudo, Japan) test kit as a screening test, followed by the Uni-Gold Recombigen® HIV-1/2 (Trinity Biotech, Ireland) assay if positive and finally confirmed by HIV1/2 STAT-PAK (Chembio Diagnostic Systems, Inc., USA). The HBsAg were tested using an ELISA kit manufactured by LabACON (Hangzhou Biotest Biotech Company, Ltd., China).
The primary endpoint was the awareness and seroprevalence of HCV. The other measures of interest included patient demographics, risk factors for HCV, and medical history.
Sample size determination
No formal sample size calculations were made because of the preliminary nature of the study .
The data were entered into an Excel 2016 spreadsheet (Microsoft Corporation, Redmond, WA, USA) and subsequently was imported into Statistical Package for the Social Sciences (SPSS) Version 22.0 (IBM Corp., Armonk, NY, USA). Statistical analyses were performed with Statistical Package for the Social Sciences (SPSS) Version 22.0 (IBM Corp., Armonk, NY, USA). The bivariate analysis was performed using a Pearson’s chi‑square test or Fisher’s exact test, whenever appropriate, to compare the demographic characteristics (e.g., age, marital status) as well as risk factors for HCV (e.g., occupational sex worker, multiple sex partner) and awareness and prevalence rates. Conditional logistic regression was employed in the multiple regression analysis to determine variables associated with awareness of HCV, while controlling for other confounding variables (such as: religion, marital status, and ownership of housing). In this analysis, the odds ratio (ORs), adjusted ORs (aORs) and confidence interval was set at 95% and p<0.05 was considered significant.
Written consent was obtained from all participants before recruitment into the study. Ethical clearance for this study was obtained from the National Health Research Ethics Committee, with registration number: NHREC/01/01/2007-23/01/2020 (approval date: January 23, 2020) in accordance with the Helsinki’s code of conduct for biomedical research involving human subjects. Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) guidance was used for reporting. Full details of the study are in the protocol .