The study samples included all individuals who donated blood from 01 January to 31 December, 2009. The study recruited 6,462 individuals in two categories; 5,878 blood replacement donors (individuals called upon to donate blood to patients) and 584 voluntary donors (individuals who willingly donated blood to the blood bank). Donor consent was obtained and confidentiality guaranteed. Clearance was also obtained from the Regional Health Directorate to use the data for the purpose of research only.

A 5^cc syringe and needle were used to bleed approximately 5 ml of blood from each donor, transferred into a vacutainer and centrifuged at 1500 rpm for 3 min to obtain serum. The Wondfo One Step Cassette Style HBsAg test kit (Guangzhou Wondfo Biotech Co. Ltd, China) was used to determine the concentration of HBsAg as being ≥1 ng/ml. A test strip was immersed in each serum. The strips were removed after 10 sec and placed on a dry clean non-absorbent table surface for 15 min after which they were visualized.

Negative test for HBV: Only one colored band appears on the control (C) region. Positive test for HBV: In addition to a pink colored control (C) band, a distinct pink colored band also appears in the test (T) region. Invalid test for HBV: A total absence of color in both regions. Also, results of recruits who could not tell their ages were eliminated.

The data obtained was double entered into a spread sheet. Descriptive analysis involved a display of summary statistics and cross tabulation of HBV, age groups and gender. We generated a binary logistic variable (as our primary dependent variable) to represent the status of HBV infection coded 0/1. The prevalence was then estimated as the proportion with infection alongside the exact binomial confidence interval. The statistical differences within age groups and between genders were tested with Pearson's chi square as well as bivariate logistic regression. A logistic regression was further used to identify the independent associations between the categories and the respective probability of HBV infection among individuals at the time of blood donation.

The data was analyzed using Stata version 11.2 (Special Edition, College Station, Texas 77845 USA) statistical package set at 95% Confidence Interval (and a p-value < 0.05 as considered statistically significant).

Out of 584 voluntary donors, 2.05% were female. The remaining 97.95% were male. Test results of 8 voluntary donors were excluded because their ages could not be determined. Of the remaining 576 voluntary donors, 10.79% tested positive for HBsAg (Table 1). Rather than expected, fewer females (12) willingly donated blood than males (572) of which 1 (8.33%) female was HBsAg + as against 62 (10.84%) males (Table 2). The age group with the highest number of donors (53.47%) was 20-29 years which also constituted the highest number of positive cases (69.35% of all positive cases) among voluntary donors (Table 3). Only 26 donors were between 40 and 60 years constituting the group with the least voluntary donors. No donor between 40 and 60 years was diagnosed HBsAg + (Table 3).

In a further analysis of a logistic regression model (Table 4), it was observed that, compared to the oldest age category for voluntary blood donors (i.e. 40-60 years), the youngest age category, 11-19 years, were 1.4 times as likely to be HBsAg+. However, the largest age category (20-29 years) were more than 2 times more likely to be HBsAg + (OR: 2.13; p = 0.009; 95%C.I. 1.2063-3.7488).

Out of the 5,878 replacement donors including only 6 (0.1%) females, 33 recruits were excluded because their ages could not be determined (Table 1). Of the remaining 5,845 recruits, 11.59% were HBsAg+. As in volunteer blood donors, the age group with the highest number of donors (49.22%) was 20-29 years. Only 77 recruits between ages of 50 and 69 years donated blood (Table 1). Generally, male donors were also more than female donors through all age groups (Table 1).

None of the 6 female replacement donors was diagnosed with HBV. All the 11.59% of total blood replacement recruits who were positive were males (Table 2). Most positive cases (60.03%) were between 20 and 29 years followed by the 30-39 year age group with 28.02% of all positive cases. The 50-69 year age group recorded the least number of recruits likewise the least number of HBsAg + cases (Table 3).

When compared to the oldest category of replacement blood donors (50-69 years), the younger age groups 11-19 and 20-29 years were more than 4 times as likely to be HBsAg + (OR: 4.04; p = 0.023; 95%C.I. 1.2118-13.4702) and (OR: 4.06; p = 0.018; 95%C.I. 1.2754-12.9532) respectively. Those aged between 40-49 were almost twice as likely to be HBsAg+, whilst the 30-39 year old category were nearly 2½ times more likely to be HBsAg + (Table 4).

There were generally more replacement donors compared to voluntary donors. The difference in prevalence between the two groups was not significant (Chi2 = 0.6117; p = 0.434)

The One Step HBsAg Test is a qualitative immunoassay that detects HBsAg in blood serum. Monoclonal and polyclonal antibodies are employed to identify HBsAg with a 1 ng/ml sensitivity. Nevertheless, the high prevalence of HBV infection (Table 1) is in concordance with some studies conducted in rural areas in developing countries where it is considered endemic [^18-22]. By using a HBsAg assay with a detection limit of 1 ng/ml, the real prevalence of HBsAg carriers might even have been underestimated. Current EIA assays are showing sensitivity of <1 ng/ml. In addition, blood donors with occult HBV infection will not be detected (HBV DNA+/HBsAg-: ~1% in Ghanaian donors) [²³].

Yet, the high prevalence obtained among blood donors who visited the hospital suggests that few people undergo immunization within the region. This is because the cost of immunization and/or testing is not within the means of the people. Most developing countries are facing multiple threats to the safety of blood supply because the cost of HBsAg screening is borne by the patients and their families who do not enjoy health coverage [¹⁷]. Generally also, there is no prevention of transmission between mother and child. High prevalence of HBsAg markers in donors who were >10 years is consistent with the concept that most primary HBV infections are transmitted either vertically or horizontally before age 10. Although most new infections occur among infants and young children, HBV-related morbidity and mortality is not immediately apparent [²⁴]. The prevalence of HBsAg among women of childbearing age in Ghana is 11.5% [²⁴]. This may also explain why low percentages of positive cases were >50 years. It is likely that by age 50 years, most carriers in the general population would have died. Low HBsAg detection in older donors might also be due to well known continuous decrease of HBsAg levels in chronic carriers over time. Most probably, HBV infection occurred before the age of 20 in these old donors. Even though there were more replacement donors, compared to voluntary donors, the difference in prevalence between the two groups was not significant (Chi2 = 0.6117; p = 0.434).

The reason for the gender imbalance, i.e. more males (6,444) than females (18), cannot be directly inferred from this data; however, within the context of the study area, more men than women are usually called to take responsibilities in representing families. The males are also more proactive and independent in decision making and volunteering than females would have the opportunity for. This probably explains the huge difference in gender. Illiteracy among the people in the region as well as poor access to relevant health information/education may also be a contributory factor to the high prevalence of the disease. The region is among those with the lowest school enrolment rate, highest dropout rate and highest illiteracy rates in the country. The disease is often attributed to supernatural or spiritual factors and/or alcoholism. This region is also prone to frequent ethnic conflicts which do not help the situation for intervention. Some socio cultural practices like polygamy may also enhance transmission particularly when a spouse either happens to contract the HBV elsewhere or a carrier who is apparently normal over years. The relatively high prevalence of HBV infection among the youth could be the result of risky lifestyles consistent with studies attributing it to unprotected sex [¹⁹,^25-27].

VBD Voluntary blood donors; RBD Replacement blood donors; Pearson chi2 for VBD (3) = 24.6740; P-value <0.001; Pearson chi2 for RBD (4) = 4.8347 P-value = 0.305

VBD Voluntary blood donors; RBD Replacement blood donors; Pearson chi2 for VBD (1) = 0.0767; Pr = 0.782; Pearson chi2 for RBD (1) = 0.7870 P-value = 0.375

VBD Voluntary blood donors; RBD Replacement blood donors; Pearson chi2 for VBD (3) = 18.1044; P-value < 0.001; Pearson chi2 for RBD (4) = 45.9008 P-value < 0.001

OR Odds Ratio; C.I. Confidence Interval