A PROJECT PROPOSAL

ON

SEROSURVEILLANCE OF HBsAg AMONG PROSPECTIVE BLOOD

DONORS IN IKARE TOWN, ONDO STATE.

By

AFE, CHARLES OLAKUNLE

AUO18AM1530

TO THE DEPARTMENT OF MEDICAL LABORATORY SCIENCE,

COLLEGE OF BASIC HEALTH SCIENCES, ACHIEVERS
UNIVERSITY, OWO, ONDO STATE.

IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE

AWARD OF BACHELOR OF MEDICAL LABORATORY SCIENCE
DEGREE (BMLS).

SUPERVISOR: PROFESSOR M.A. MUHIBI

FEBUARY, 2023
BACKGROUND TO THE STUDY

Blood donation is the process of drawing blood from a healthy volunteer and

collecting and storing it for transfusion into a patient with a matched blood group. Blood

transfusion plays an important role in saving patient lives and stabilizing their medical

conditions, but transfusion-transmitted diseases, via blood-borne pathogens, remain a medical

concern (Alqahtani et al., 2021).

The most notable transfusion-related risks worldwide include hepatitis. Hepatitis is a

serious liver disease caused by a group of viruses. It is characterized by inflammation of the

liver with a presence of inflammatory cells which are triggered by the infection and can lead

to severe complication such as fibrosis or cirrhosis, which might be fatal (Karnsakul and

Schwarz, 2017).

Hepatitis B virus is a DNA virus which belongs to the hepadna virus family. Viruses

in this family are unique in that they replicate from an RNA file involving reverse

transcriptase and they are liable to developing mutant strains. It affects the liver resulting into

inflammatory conditions (Odiabara et al., 2020).

One-third of the world’s population (more than two billion people) are infected with

HBV, which therefore poses a major health challenge; however, excluding

immunocompromised individuals, 95% of adult hepatitis B patients recover with minimal or

no clinical care. In contrast, the recovery rate in children is below 10% (WHO, 2022).

Hepatitis B is a viral infection that attacks the liver and can cause both acute and

chronic disease. Acute hepatitis B refers to newly acquired infection with an average

incubation period of about 90 days. At this stage over 95% of infected persons are

asymptomatic ,the disease may resolve after weeks to months mostly in adults without

treatment, the remaining 5% develop severe life threatening form of acute (fulminant) or

chronic hepatitis disease (Fasakin et al., 2022).

 People who recover from the virus become immune and cannot get infected again.

The first serologic markers that can be detected at this stage are hepatitis B surface antigen

(HBsAg) and antibodies to hepatitis B core antigen (anti-HBcAg) and are helpful for the

diagnosis of the infection (Ojevwe et al., 2021).

Chronic hepatitis B infection is infection that has lasted more than 6 months and may

never go away. WHO estimates that 296 million people were living with chronic hepatitis B

infection in 2019, with 1.5 million new infections each year (WHO, 2022). In 2019, hepatitis

B resulted in an estimated 820 000 deaths, mostly from cirrhosis and hepatocellular

carcinoma (primary liver cancer) (WHO, 2022).

Hepatitis B virus can be transmitted through both horizontal and vertical (perinatal)

routes. The infection occurs through contact with infected blood, blood products, unprotected

sex with an infected person, or shared needles; additionally, an infected mother can transmit

HBV to her baby before, during, or after childbirth (Karnsakul and Schwarz, 2017; Sanni and

Olorunfemi, 2019).

The risk of hepatitis B virus infection through unsafe blood is exceptionally high

particularly in sub-Saharan Africa (Ezeonu et al., 2019). It is reported that relatively long

viremic periods during which individuals remain asymptomatic and efficient transmission

through contaminated blood products could contaminate blood supply and pose high risk of

serious disease manipulation to recipients (Agbesor et al., 2015). Not only does hepatitis B

virus infection cause a high morbidity, its effects also stress clinical resources and can also

have economic consequences. The pathogenesis of hepatitis B virus infection can be

enhanced by several factors which include the age at infection, level of viral replication and

loss of immune responses (Odiabara et al., 2020).

Nigeria is classified among the group of countries highly endemic for HBV infection.

About 75% of the Nigerian population is reportedly to have been exposed to HBV at one time
or the other in life (Ojevwe et al., 2021). The infection has reached hyper endemic level in

Nigeria with estimated seroprevalence of hepatitis B surface antigen (HbsAg) of 10 – 40% it

has been observed that the possibility of contacting the virus in Nigeria is high due to low

vaccination rate and the fact that about 75% of population will be exposed (Odiabara et al.,

2020). Previously, various seroprevalence rates for HBsAg have been reported among blood

donors in different part of Nigeria with 5.9% in Ibadan (Afolabi et al., 2015), 17.5% in Abuja

(Agbesor et al., 2015) and 10.5% in Ogun state (Sanni and Olorunfemi, 2019).

STATEMENT OF PROBLEM

Hepatitis B is one of the most notable transfusion-related risks in Nigeria. Blood has

the highest concentration of the virus, so transfusion of infected blood is the surest way to get

infected. Approximately nine in ten Nigerians who live with chronic HBV are unaware of

their infection status (Ajuwon et al., 2021). Hepatitis B virus infection cause a high

morbidity and mortality, its effects also stress clinical resources and can also have economic

consequences.

JUSTIFICATION OF STUDY

Transmission of hepatitis B has long been identified as a major public health issue in

the tropical and subtropical countries such as Nigeria. Hepatitis B constitutes a serious threat

to human race due to its high rate of fatality. The improved screening and testing of

prospective blood donors will significantly reduce transfusion transmitted diseases in Nigeria.

The disease trends and blood assessment can better be done by the study of serosurveillance

rate of hepatitis B in prospective blood donors and indexed accurately in general population.

This study will be carried out to determine the serosurveillance of HBsAg among prospective

blood donors in State Specialist Hospital, Ikare Town, Ondo State.

AIM

The aim of this study is to determine the seroprevalence of HBsAg among prospective blood

donors in State Specialist Hospital, Ikare Town, Ondo State.

RESEARCH OBJECTIVES

Specific objectives of the study are the following:

i. To determine the prevalence of Hepatitis B among prospective blood donors.

ii. To determine the serosurveillance of HBsAg among prospective blood donors.

iii. To determine the association between demographic/biologic characteristics and

the outcome of the serosurveillance of HBsAg.

iv. To identify the risky health behaviours that may predispose to HBV infections

among prospective blood donors.

RESEARCH HYPOTHESIS

Null Hypothesis (H0): There is no serosurveillance of HBsAg among prospective blood

donors.

Alternative Hypothesis (H1): There is serosurveillance of HBsAg among prospective blood

donors.

SIGNIFICANCE OF STUDY

The research work shall provide useful information on the serosurveillance of HBsAg

among prospective blood donors. It will provide useful information about prospective blood

donor’s immunity due to past hepatitis B infection or vaccination. The research work shall

provide useful information on the prevalence of Hepatitis B among prospective blood donors

in Ikare Town, Ondo State.

 LITERATURE REVIEW

HEPATITIS B VIRUS

Hepatitis B virus (HBV) is a partially double-stranded DNA virus, a species of the

genus Orthohepadnavirus and a member of the Hepadnaviridae family of viruses. This virus

causes the disease hepatitis B (Sanni and Olorunfemi, 2019). The virus is divided into four

major serotypes (adr, adw, ayr, ayw) based on antigenic epitopes present on its envelope

proteins. These serotypes are based on a common determinant (a) and two mutually exclusive

determinant pairs (d/y and w/r). The viral strains have also been divided into ten genotypes

(A–J) and forty subgenotypes according to overall nucleotide sequence variation of the

genome (Hundie et al., 2017). The genotypes have a distinct geographical distribution and are

used in tracing the evolution and transmission of the virus. Differences between genotypes

affect the disease severity, course and likelihood of complications, and response to treatment.

The serotypes and genotypes do not necessarily correspond (Wang et al., 2019).

Hepatitis B is a serious liver infection caused by the hepatitis B virus (HBV). It is a

major global health problem. For some people, hepatitis B infection becomes chronic,

meaning it lasts more than six months. Having chronic hepatitis B increases the risk of

developing liver failure, liver cancer or cirrhosis (a condition that permanently scars of the

liver). Most adults with hepatitis B recover fully, even if their signs and symptoms are severe.

Infants and children are more likely to develop a chronic (long-lasting) hepatitis B infection.

A vaccine can prevent hepatitis B, but there is no cure if an individual have the condition. If

an individual is infected, taking certain precautions can help prevent spreading the virus to

others (Terrault et al., 2018).

Hepatitis B virus (HBV) attacks liver parenchyma cells (hepatocytes) and cause

hepatitis which may be acute or chronic. Although majority of individuals recover from the
acute infection, those who progress to chronic hepatitis B infection are at great risk of

developing serious complications such as cirrhosis, hepatocallular carcinoma, liver failure

and eventual death (Yakubu et al., 2016).

MORPHOLOGY AND STRUCTURE OF HCV

Hepatitis B virus is a member of the Hepadnavirus family. The virus particle, called

Dane particle (virion), consists of an outer lipid envelope and an icosahedral nucleocapsid

core composed of protein. The nucleocapsid encloses the viral DNA and a DNA polymerase

that has reverse transcriptase activity similar to retroviruses (Menéndez-Arias et al., 2014).

The outer envelope contains embedded proteins which are involved in viral binding of, and

entry into, susceptible cells. The virus is one of the smallest enveloped animal viruses with a

virion diameter of 42 nm, but pleomorphic forms exist, including filamentous and spherical

bodies lacking a core. These particles are not infectious and are composed of the lipid and

protein that forms part of the surface of the virion, which is called the surface antigen

(HBsAg), and is produced in excess during the life cycle of the virus (Kmet Lunacek et al.,

2017).

HEPATITIS B GENOME

HBV is a partially double-stranded DNA virus of 3.2 kilobases, and it transforms

from pregenomic ribonucleic acid (RNA) to DNA by reverse transcription during its life

cycle. The genome consists of an outer lipid envelope and inner nucleocapsid core encoded

by four overlapping open reading frames, named C, X, P, and S (McNaughton et al., 2019).

Although it is known as a virus with high replication ability, due to the absence of the

proofreading reverse transcriptase enzyme, the naturally occurring mutations may arise in

different genome regions. These regions may encode for polymerase, surface antigen,

core/precore promoter, and comprise the X genes that significantly influence HBsAg

expression and progression of HCC (Arikan et al., 2019). Additionally, due to the complete
overlapping of pol and S genes, drug resistance and nucleos(t)ide resistance mutations

occurring in the pol gene can lead to changes in its product HBsAg (Kırdar et al., 2019).

The mutations in the gene C that encode for precore and core proteins are

significantly correlated with liver disease progression in CHB patients (Al-Qahtani et al.,

2018). The changes in the amino acid sequences: W28*, G29D, G1896A, G1899A, G1862T

in the precore proteins that affect HBeAg, and F24Y, E64D, E77Q,A80I/T/V, L116I, E180A

in the core proteins mutations are commonly identified and related to clinical severity (Kim

et al., 2016).

GENOTYPES

HBV is a hepatotropic virus and has a partially doublestranded, relaxed circular DNA

genome containing about 3.2 kilobases. Due to the lack of proofreading activity of its reverse

transcriptase, the mutation rate of HBV is very high, with an estimated nucleotide

replacement rate of 1.4 to 3.2 x 10 -5 per site per year (Liu et al., 2021). The high error rate of

HBV reverse transcriptase during viral replication results in frequent nucleotide substitutions,

leading to genotype, subgenotype, and quasispecies diversity (Zhang et al., 2016). In 1988,

HBV could be divided into 4 genotypes based on genome nucleotide variation that is greater

than 8%. Since then, at least 10 genotypes (A–J) have been identified. Genotypes A, B, C, D,

and F have been further split into subgenotypes with 4% genome divergence (Arikan et al.,

2016). In addition to the full-length HBV sequences, the S gene is also suitable for

genotyping because it is more highly conserved than other genes in the HBV genome (Liu et

al., 2021).

In recent years, HBV genotypes and subgenotypes have attracted increasing attention

since they influence the activity and outcome of chronic HBV infection. For example,

genotype A is common in CHB patients, while genotype D is more prevalent in patients with
acute hepatitis. Genotype D is the most frequent genotype in patients with acute liver failure

(Anastasiou et al., 2019). Compared to genotype B, genotype C is associated with late

hepatitis B e antigen (HBeAg) seroconversion and advanced liver disease (Zhang et al.,

2016).

A study has shown that the frequency of genotype C was higher than genotype B in

CHB/HBV-related liver cirrhosis and hepatocellular carcinoma (HCC) patients, while

genotype B was more frequent in asymptomatic carrier and liver failure patients (Wu et al.,

2017). Genotype H has a low level of HBV DNA and HBeAg, and has been associated with

occult HBV and less severe liver disease (Sozzi et al., 2018). Subgenotypes A1 and D1 have

been associated with lower levels of HBV DNA and early HBeAg seroconversion (Ozaras et

al., 2015). Subgenotype F1 has been associated with a higher risk of HCC compared with

genotypes A, B, and D (Arikan et al., 2016). HBV genotypes are not only associated with

clinical progression of disease but are also related to interferon treatment responses. In a

word, genotype determination in HBV infection is important for disease progression

estimations and the design of optimal antiviral treatment plans (Liu et al., 2021).

Many studies have reported that different genotypes and subgenotypes show different

geographical distributions. Genotype A is mainly found in Europe (where the main

subgenotype is A2) and Africa (A1). Genotypes B and C are common in Asia. Genotype D is

prevalent in Africa, Europe, the Mediterranean region, and India. Genotype E is found in

West and South Africa. Genotype F is dominant in South and Central America. Genotype G

has been reported in France and the United States. Genotype H is present in Mexico and

Central America. Genotype I has been identified in Vietnam and Laos. The newest HBV

genotype, genotype J, has been reported in the Ryukyu Islands of Japan (Liu et al., 2021).

The distribution of genotypes not only varies between countries, but also between

geographical regions within countries. In China, the main genotypes are genotypes B and C,
with genotype B predominating in the central region and genotype C predominating southern

and northern regions (Wu et al., 2017; Liu et al., 2021).

TRANSMISSION

The spread of Hepatitis B virus occurs most often through vertical transmission from

mother to child during birth and delivery. HBV can also be spread through contact with blood

or other bodily fluids during sexual intercourse with an infected partner. It is also spread

through needles shared with infected persons or exposure to sharp objects. Needles of any

kind can be a risk if they are not single use or are not properly sanitized, preferably in an

autoclave. This includes needles used at tattooing and body piercing parlors (CDC, 2022).

Furthermore, Hepatitis B virus can also be spread through sharing earrings and other

body piercing jewellery (WHO, 2022). It is also spread through haemodialysis units that have

been used by HBeAg positive patients. Because HD units usually treat multiple patients at a

time, contamination of patients’ blood can occur. Incidences of HBV infection through HD

units is at 1% in the United States. Healthcare staff are also at an increased risk of infection

(Garthwaite et al., 2019).

Transmission of HBV can be limited by administering the Hepatitis B vaccine. In

areas where the virus is endemic, vaccines are limited, especially in rural areas where medical

clinics are sparse (Terrault et al., 2018). Although HBV can be infectious on surfaces for up

to seven days, it is not spread through breastfeeding, sharing eating utensils, hugging, kissing,

holding hands, coughing, or sneezing. Unlike other Hepatitis viruses, HBV is not spread by

contaminated food or water. However, living with a person infected with Hepatitis B virus

increases your risk of contracting the virus (CDC, 2022).

CLINICAL SIGNS AND SYMPTOMS

The clinical course of acute hepatitis B is indistinguishable from that of other types of

acute viral hepatitis. The incubation period typically ranges from 60 to 90 days. Clinical signs
and symptoms occur more often in adults than in infants or children; infants and young

children usually are asymptomatic. Approximately 50% of adults who have acute infections

are asymptomatic (Ojevwe et al., 2021).

The preicteric, or prodromal, phase from initial symptoms to onset of jaundice usually

lasts 3 to 10 days. It is nonspecific and is characterized by abrupt onset of fever, malaise,

anorexia, nausea, abdominal discomfort, and dark urine beginning 1 to 2 days before the

onset of jaundice. The icteric phase is variable but usually lasts from 1 to 3 weeks and is

characterized by jaundice, light or gray stools, hepatic tenderness, and hepatomegaly

(splenomegaly is less common). During convalescence, malaise and fatigue may persist for

weeks or months, while jaundice, anorexia, and other symptoms disappear (Liang et al.,

2015).

ACUTE HBV INFECTIONS

Most acute HBV infections in adults result in complete recovery with elimination of

HBsAg from the blood and the production of anti-HBs, creating immunity to future infection.

In contrast, as many as 90% of HBV infections in infants progress to chronic infection. While

most acute HBV infections in adults result in complete recovery, fulminant hepatitis occurs in

about 1% to 2% of acutely infected persons (Ojevwe et al., 2021). Although the

consequences of acute HBV infection can be severe, most of the serious complications

associated with HBV infection are due to chronic infection (Liang et al., 2015).

CHRONIC HBV INFECTION

The proportion of people with acute HBV infection who progress to chronic infection

varies with age and immune status. As many as 90% of infants who acquire HBV infection

from their mothers at birth or in infancy become chronically infected. Of children who

become infected with HBV between age 1 and 5 years, 30% to 50% become chronically
infected. The risk of acquiring chronic HBV infection when infected during adulthood is

approximately 5% (Eko Mba et al., 2018).

Persons with chronic infection are often asymptomatic and may not be aware they are

infected; however, they are capable of infecting others and have been referred to as carriers.

Chronic infection is responsible for most HBV-related morbidity and mortality, including

chronic hepatitis, cirrhosis, liver failure, and HCC. Approximately 25% of persons who

become chronically infected during childhood and 15% of those who become chronically

infected after childhood will die prematurely from cirrhosis or liver cancer (Terrault et al.,

2018).

HEPATITIS B SURFACE ANTIGEN (HBSAG)

HBsAg is the most used test for diagnosing acute HBV infections or detecting

carriers. HBsAg can be detected as early as 1 or 2 weeks and as late as 11 or 12 weeks after

exposure to HBV. The presence of HBsAg indicates that a person is infectious, regardless of

whether the infection is acute or chronic. Transient HBsAg positivity can occur up to 18 days

following vaccination (up to 52 days among hemodialysis patients) and is clinically

insignificant (Karnsakul and Schwarz, 2017).

HEPATITIS B PREVENTION STRATEGIES

HepB vaccination is the mainstay of hepatitis B prevention efforts. A comprehensive

strategy to eliminate HBV transmission includes universal vaccination of infants beginning at

birth, routine vaccination of previously unvaccinated children less than age 19 years, and

vaccination of adults at risk for HBV infection, including those requesting protection from

HBV without acknowledgement of a specific risk factor. It also includes universal testing of

pregnant women for HBsAg to identify newborns who require immunoprophylaxis for

prevention of perinatal infection and to pregnant women who can benefit from antiviral

therapy to reduce perinatal transmission (Abdella et al., 2018; Van Damme et al., 2018).
BLOOD DONATION

Blood donation occurs when people voluntarily have blood drawn and used for

transfusion and/or made into biopharmaceutical medications by a process called

fractionalization (separation of whole blood components). Donation may be of whole blood

or of specific blood component directly (the later called aphresis). Blood banks often

participate in the collection process as well as the procedure that follows it including storage

(Burgdorf et al., 2017). In most countries blood donors are unpaid volunteers who donate

blood for community supply. In poorer countries most time, donors usually give blood to

their relatives and friends who need transfusion (directed donation) with many donors

donating as an act of charity. Individual donor can also donate for their own future use

(autologous donation) (Sanni and Olorunfemi, 2019).

Before donation, potential donors are evaluated for anything that might make their

blood unsafe to use. The screening includes testing for diseases that can be transmitted by

blood transfusion including HIV, Hepatitis B and Hepatitis C. The donor also answers

question about medical history and take a short physical examination to make sure the

donation is not hazardous to his or her health. This has greatly helped in reducing blood

transfusion induced Hepatitis B and C infections (Afolabi et al., 2015; Ikobah et al., 2016).

BLOOD DONORS

There are several types of blood donors, such as;

i. Voluntary nonremunerated donors (VNRDs): are defined as blood donors who donate

blood without any relationships to the recipients or remuneration.

ii. Family replacement donors (FRDs): are blood donors who donate blood without

remuneration but for their family members or friends.

 iii. Paid donors (PDs): are those blood donors who donate blood after receipt of

remuneration (Yakubu et al., 2016).

BLOOD TRANSFUSION AND HEPATITIS B VIRUS

Blood donation is an important, lifesaving intervention. Donations from non-

remunerated volunteers have been proven to the safest. Among the various routes of

transmission of HBV infection, blood transfusion transmission represents a deliberate process

of transmission that can be avoided even with the slightest vigilance (Ezeonu et al., 2019).

Although, the incidence of transfusion transmitted hepatitis B has been gradually controlled

within the intervening period, the available records show that HBV remains yet the viral

infection with the greatest risk of transmission through blood transfusion especially in

developing countries (Musa et al., 2015).

An obviously high variability in the prevalence of HBV is seen across the countries,

with higher incidence and prevalence observed in developing nations such as 1.6%–7.7% in

Brazil, 19.6% in Egypt and 2-10% across India (La Torre and Saulle, 2016). In Nigeria, the

pooled prevalence of HBV is reported to be about 13.6% (Ikobah et al., 2016).

Blood has the highest concentration of the virus, so transfusion of infected blood is

the surest way to get infected (Obionu and Uzochukwu, 2013). Chronic asymptomatic

carriers are reservoirs and are potential source of new infections. Hepatitis B virus may be

circulating in the blood stream for weeks to months, even years without causing an overt

symptom; therefore, apparently healthy carriers may pass blood donation selection criteria

and accepted as suitable to donate (Odiabara et al., 2020).

Even though there are routine tests for demonstrating the presence or otherwise of this

infection, the greatest risk are blood donations made in the infectious window period, which

is the time between development of infectious viraemia and/or reactivity to routine

serological or nucleic acid technology (NAT) based screening tests (Ezeonu et al., 2019). The
safety of blood can be guaranteed by application of assay technologies with high specificity

and sensitivity so as to be able to identify all true positive blood units and true negative blood

units (Yakubu et al., 2016; Abdella et al., 2018).

Screening of blood donors is a critical issue as the outcome of the test if flawed for

whatever reason can result in serious consequences for either the blood service or the blood

donor. False positive result can lead to a larger number of blood donors being deferred, while

a false negative testing may jeopardize blood safety (Ezeonu et al., 2019). In Nigeria, the

prevalence of HBV infection has been speculated to vary with regard to the screening method

used with prevalence rates varying from 12.3% by enzyme linked immunosorbent assay,

17.5% by immuno-chromatography and 13.6% by HBV DNA polymerase chain reaction

(Musa et al., 2015).

 MATERIALS AND METHOD

STUDY DESIGN

The study design used will be a cross-sectional study

STUDY AREA

The research will be carried out among prospective blood donors in Ikare Town, also

called Ikare-Akoko town, Ondo State, southwestern Nigeria. It lies in the Yoruba Hills, at the

intersection of roads from Owo, Okene, Kabba and Ado-Ekiti. The geographical coordinates

are 7˚ 31ˈ 0̎ North, 5˚ 45ˈ 0̎ East. An agricultural trade centre for the local Yoruba people.

Ikare has an estimated population of 1.1 million and an area of 540km 2. Ikare has the tropical

savanna climate. In Ikare, the wet season is warm, oppressive, and overcast and the dry

season is hot, muggy and partly cloudy.

ETHICAL APPROVAL AND INFORMED CONSENT

Ethical approval will be obtained from Research Ethics Committees of State

Specialist Hospital, Ikare Town, Ondo State. Informed consents of the prospective blood

donors will be obtained before enrolment after due explanation of the aims and procedures of

the research and the participants will be enrolled consecutively. All eligible participants will

sign an informed consent form before the administration of questionnaire and sample

collection. Privacy and confidentiality of the entire participants will be guaranteed by coding

the data to ensure anonymity of the participants.

INCLUSION AND EXCLUSION CRITERIA

Prospective blood donors at the time of this study from the target population at State

Specialist Hospital, Ikare Town that are adult between the ages of 18-65years, weight of 50kg
and above except if a smaller (250ml) blood donor bag is to be used, hemoglobin

concentration above 13g/dl in men, >12g/dl in women, minimum donation interval of 12

weeks and maximum of three donations per year, and will give informed consent will be

included.

Individuals from the target population that will not give their consent, pregnant

women, extremely anemic patients (<70g/L), and any individual with a cardiovascular,

respiratory, gastrointestinal, renal, or epileptic disorder or those that are currently on

medication will be excluded.

DATA COLLECTION

A self-administered simplified questionnaire will be used to collect information on

demographic and biological variables. The questionnaire will be checked for errors and a trial

run of the questionnaire will be done to resolve any vague areas.

SAMPLE SIZE DETERMINATION

A sample is a representative subgroup of the population that meets researchers’

criteria. A minimum sample size was obtained using sample size calculation for proportion
(formula as shown below)

The minimum sample size required for this study was estimated based on the prevalence of

7.4% for HBV in prospective blood donors in Akure, Ondo State as reported by (Shittu et al.,

2014).

Where:
n= The minimum sample size required

P= The prevalence of 7.4% for HBV in prospective blood donors = 7.4% = 0.074

q= 1-p = 1-0.074 = 0.926

Z= corresponds to significance level (1.96 for 0.05)

d= absolute error or precision= 5% (0.05)

Calculation:

n= (1.96)2 × 0.074× (1-0.074)/ (0.05)2

n= 3.8416 × 0.074× 0.926/0.0025

n= 0.2632/0.0025

n= 105.3

Thus, the estimated required minimum sample size for this study is 105 participants

SAMPLE COLLECTION/ ANALYSIS

Five (5) milliliters of venous blood will be collected using a plain bottle from the

participants after obtaining written informed consent and filling the questionnaire from each

participant. The blood collected will be separated immediately by centrifugation at 3000rpm

for 15minutes using a laboratory bench centrifuge (Searchtek Instruments). The sera will be

stored at -30oC for HBsAg screening using enzyme linked immunosorbent (ELISA) assay kit.

ANALYTICAL METHODS

Presence of HBsAg in serum will be determined using enzyme linked immunosorbent

assay (ELISA) as described by (Ezeonu et al., 2019)

STATISTICAL ANALYSIS

The data generated will be coded, entered, validated and analysed using Statistical

Package for Social Science (SPSS) version 25.0. Subgroup analysis will be done and the

difference will be compared using chi-square. The level of significance will be taken at 95%

confidence interval and fixed at P<0.05.

EXPECTED OUTCOMES

1. The prevalence of HBV among prospective blood donors in Ikare Town

2. The serosurveillance of HBsAg among prospective blood donors in Ikare Town

3. The association between demographic, biologic characteristics and the outcome of

the HBsAg screening.

REFERENCES

Abdella, Y., Riedner, G., Hajjeh, R. & Sibinga, C.T.S. (2018). Blood transfusion and
hepatitis: What does it take to prevent new infections? Eastern Mediterranean Health
Journal, 24, 595–597.

Afolabi, A. Y., Oladipo, E. K., and Fagbami, A. H. (2015). Prevalence of hepatitis B virus
among potential blood donors in Ibadan, Nigeria. International Research Journal of
Medicine and Medical Sciences, 3(2), 40-45.
Agbesor, N. I., Amala, S. E., and Zaccheaus, A. J. (2015). Hepatitis B virus profile among
blood donors in Federal Capital Territory Abuja, Nigeria. International Journal of
Science Research, 5(7): 2319.
Ajuwon, B.I., Yujuico, I., Roper, K., Richardson, A., Sheel, M. & Lidbury, B.A. (2021).
Hepatitis B virus infection in Nigeria: a systematic review and meta-analysis of data
published between 2010 and 2019. BMC Infectious Diseases, 21, 1120 – 1135.

Al-Qahtani, A. A., Al-Anazi, M. R., Nazir, N., Abdo, A. A., Sanai, F. M., Al-Hamoudi, W.
K., Alswat, K. A., Al-Ashgar, H. I., Khan, M. Q., & Albenmousa, A. (2018). The
correlation between hepatitis B virus Precore/core mutations and the progression of
severe liver disease. Frontiers in Cellular and Infection Microbiology, 8, 355.

Alqahtani, S. M., Alsagaby, S., Mir, S. A., Alaidarous, M., Bin Dukhyil, A., Alshehri, B.,
Banawas, S., Alturaiki, W., Alharbi, N. K. & Azad, T. A. (2021). Seroprevalence of
Viral Hepatitis B and C among Blood Donors in the Northern Region of Riyadh
Province, Saudi Arabia. Healthcare, 9(934), 1-11.

Anastasiou, O. E., Widera, M., & Westhaus, S. (2019). Clinical outcome and viral genome
variability of hepatitis B virus-induced acute liver failure. Hepatology, 69, 993–1003.

Arikan, A., Sayan, M., Sanlidag, T., Suer, K., Akcali, S. & Guvenir, M. (2019). Evaluation of
the pol/S gene overlapping mutations in chronic hepatitis B patients in Northern
Cyprus. Poland Journal of Microbiology, 68(3), 317–322.

Burgdorf, K. S., Simonsen, J., Sunddby, A., Rostgaard, K., Pedersen, O. B., Serensen, E.,
Nielsen, K. R., Bruun, M. T., Frisch, M., Edgren, G., Erikstrup, C., Hjalgrim, H. &
Ullum, H. (2017). Socio-demographic charateristics of Danish blood donors, PloS
ONE, 12(2), 1-6.

Centers for Disease Control and Prevention, (2022)."Hepatitis B | Disease Directory |

Travelers' Health | CDC" Available from
https://wwwnc.cdc.gov/travel/diseases/hepatitis-b. Retrieved 25th January, 2023.

Eko Mba, J. M, Bisseye, C., Ntsame Ndong, J. M., Mombo, L. E., Bengone, C. & Mouelet
Migolet, G. (2018) Prevalent hepatitis B surface antigen among first-time blood
donors in Gabon. PLoS ONE, 13(4), 1-10.
Ezeonu, C. M., Garba, S. A., Adabara, N. U. & Kuta, F. A. (2019). Prevalence of hepatitis B
virus infections among blood donors in Federal Capital Territory, Abuja, Nigeria.
African Journal of Clinical and Experimental Microbiology, 20(3), 182-194.

Fasakin, K. A., Muhibi, M. A., Ibijola, A. A., Ajayi, O. D., Onyema, M. I., Obazee, Y.,
Adeniyi, A. A., Ajetunmobi, W. A., Adebara, I. O., Adebayo, A., Okunlola, A. I.,
Yusuf, M., Awoyinka, B. S., Ajetunmobi, O. A., Bakare, A., Adeyemo, O. T.,
Adewara, O. E., Anyahara, N., Onyema, C. E., Bolaji, O. B., Ayankunle, O. M.,
Aduloju, O. P., Ogundare, E. O., Babatola, A. O., Adefisan, A. S., Olofinbiyi, B. A. &
Jeremiah Z. A. (2022). Sero-Epidemiology of Transfusion Transmissible Hepatitis B,
C and E among Blood Donors in Ekiti, Southwestern Nigeria: A Cross-sectional
Study. Journal of Advances in Medicine and Medical Research, 34(13), 61-72.

Garthwaite, E., Reddy, V., Douthwaite, S., Lines, S., Tyerman, K. & Eccles, J. (2019).
"Clinical practice guideline management of blood borne viruses within the
haemodialysis unit". BMC Nephrology, 20(1), 388.

Hundie, G. B., Stalin Raj, V., Gebre M. D., Pas, S. D., Koopmans, M. P., Osterhaus, A. D.
(2017). "A novel hepatitis B virus subgenotype D10 circulating in Ethiopia". Journal
of Viral Hepatitis, 24(2), 163–173.
Ikobah, J., Okpara, H., Elemi, I., Ogarepe, Y., Udoh, E. & Ekanem, E. (2016). The
prevalence of hepatitis B virus infection in Nigerian children prior to vaccine
introduction into the National Programme on Immunization schedule. Pan African
Medicine Journal, 23, 128.

Karnsakul, W., & Schwarz, K. B. (2017). Hepatitis B and C. Pediatric Clinics of North
America, 64, 641–658.

Kim, H., Lee, S. A., Do, S. Y., & Kim, B. J. (2016). Precore/core region mutations of
hepatitis B virus related to clinical severity. World Journal of Gastroenterology,
22(17), 4287–4296.

Kırdar, S., Yaşa, M. H., Sayan, M. & Aydın, N. (2019). [HBV pol/S gene mutations in
chronic hepatitis B patients receiving nucleoside/nucleotide analogues treatment] (in
Turkish). Mikrobiyoloji Bulteni, 53(2), 144–155.

Kmet Lunacek, N., Poljak, M., Meglic-Volkar, J., Rajter, M., Prah, J., Lesnicar, G., Selic
Kurincic, T., Baklan, Z., Remec, T., & Pal, E. (2017). Epidemiological, virological
and clinical characteristics of hepatitis B virus genotypes in chronically infected
persons in Slovenia. Hepatitis Monthly, 17(3), 1-8.

La Torre, G., & Saulle, R. (2016). Risk factors from HBV infection among blood donors: A
systematic review. Asia Pacific Journal of Tropical Biomedicine, 6(4): 344–349.

Liang, T. J., Block, T. M., McMahon, B. J., Ghany, M. G., Urban, S., Guo, J. T., Locarnini,
S., Zoulim, F., Chang, K. M., & Lok, A. S. (2015). Present and future therapies of
hepatitis B: from discovery to cure. Hepatology. 62(6), 1893–1908.

Liu, Z., Zhang, Y., Xu, M., Li, X., & Zhang, Z. (2021). Distribution of hepatitis B virus
genotypes and subgenotypes: a meta-analysis. Medicine, 100(50), 1- 9.
McNaughton, A. L., D’Arienzo, V., Ansari, M. A., Lumley, S. F., Littlejohn, M., Revill, P.,
McKeating, J. A., & Matthews, P. C. (2019). Insights from deep sequencing of the
HBV genome-Inique,tiny, and misunderstood. Gastroenterology, 156(2), 384–399.

Menéndez-Arias, L., Álvarez, M. & Pacheco, B. (2014). "Nucleoside/nucleotide analog

inhibitors of hepatitis B virus polymerase: mechanism of action and resistance".
Current Opinion in Virology, 8, 1–9.

Musa, B. M., Bussell, S., Borodo, M. M., Samaila, A. A., and Femi, O. L. (2015). Prevalence
of hepatitis B virus infection in Nigeria, 2000-2013: A systematic review and meta-
analysis. Nigerian Journal of Clinical Practice, 18(2), 162-172.

Obionu, C. N., & Uzochukwu, B. S. C. (2013). National Survey of Viral Hepatitis B and C
Infection in Nigeria. Nigeria: Federal Ministry of Health; 1-25.

Odiabara, K. K., Nwachuku, E. O., Boisa, N. & Ebirien-Agana, S.B. (2020). Determination
of Hepatitis B Virus Carriers among Voluntary Blood Donors in Some Parts of
Nigeria. Scholars Journal of Applied Medical Sciences, 8(3), 775-782.

Ojevwe, D. D., Fajimi, V. O., Ifemeje, O. C., Ali, H. A., Gana, D. F., Abdulsalam, J. O.,
Adegoke, A. S., Agaga, F., & Maduka, M.K. (2021). Prevalence of hepatitis B
infection among voluntary donor in Wuse District Hospital, Abuja. GSC Biological
and Pharmaceutical Sciences, 14(01), 109–115.

Ozaras, R., Inanc Balkan, I., Yemisen, M. & Tabak, F. (2015). Epidemiology of HBV
subgenotypes D. Clinics and Research in Hepatology Gastroenterology, 39, 28–37.

Sanni, T.A. and Olorunfemi E.A. (2019). Prevalence and Risky Health behaviours Associated
with Hepatitis B and C Infection among Blood Donors in Ogun State, Southwest,
Nigeria. Texila International Journal of Clinical Research, 2, 1-9.

Shittu, M. O., Adekola, A.A., Ajao, K.O., Adeniji, T.W. & Awe, C.O. (2014).
Seroprevalence of Hepatitis B Surface Antigenemia and Hepatitis C Virus among
Intending Blood Donors at Mother and Child Hospital, Akure, Nigeria. International
Journal of Medicine and Medical Sciences, 47(2), 1-5.

Sozzi, V., Shen, F. & Chen, J. (2018). In vitro studies identify a low replication phenotype for
hepatitis B virus genotype H generally associated with occult HBV and less severe
liver disease. Virology, 519, 190–196.

Terrault, N. A., Lok, A. S. F., McMahon, B. J., Chang, K. M., Hwang, J. P., Jonas, M. M.,
Brown, R. S. Jr., Bzowej, N. H. & Wong, J. B. (2018). Update on prevention,
diagnosis, and treatment of chronic hepatitis B: AASLD 2018 hepatitis B guidance.
Hepatology, 67(4), 1560–1599.

Van Damme, P., Ward, J. & Shouval, D. (2018). Hepatitis B Vaccines. In: Plotkin S,
Orenstein, W, Offit P.,eds. Plotkin’s Vaccines. 7th ed. Elsevier; 342–374.
Wang, W., Shu, Y., Bao, H., Zhao, W., Wang, W., Wang, Q., Lei, X., Cui, D. & Yan, Z.
(2019). Genotypes and hot spot mutations of hepatitis B virus in northwest Chinese
population and its correlation with disease progression. BioMed Research
International 10, 1–9.

World Health Organisation, (2022). "Hepatitis B" Available from https://www.who.int/news-

room/fact-sheets/detail/hepatitis-b. www.who.int. Retrieved 2 January,2023.

Wu, X., Wei, S. F., Xu, N., Su, Q., Li, X. & Zhang, Z. H. (2017). Meta-analysis on
distribution of hepatitis B virus genotypes and related clinical outcomes in China [in
Chinese]. Journal of Clinical Hepatology, 2017, 271–275.

Yakubu, A., Sahabi, D. M., Umar, A., Saidu, Y., & Magaji, U.F. (2016). Prevalence of
HBsAg among Prospective Blood Donors and Pregnant Women in Kebbi State,
Nigeria. Nigerian Journal of Basic and Applied Sciences, 24(1), 57-63.

Zhang, Z. H., Wu, C. C., Chen, X. W., Li, X., Li, J., & Lu, M. J. (2016). Genetic variation of
hepatitis B virus and its significance for pathogenesis. World Journal of
Gastroenterology, 22, 126–144.

APPENDIX I

QUESTIONNAIRE

SECTION A: SOCIO-DEMOGRAPHIC FACTORS OF THE PARTICIPANTS

Please kindly complete this questionnaire; all information will be kept confidential

Date:………………………………..

Serial Number:…………………………

Age
15-24 25-34 35-44 45-54 55-64 >64yrs
Sex
Male Female
Occupation

Educational level
Non-formal
 Primary
Secondary
Tertiary
Marital status
Single Married Divorced Widowed
Tribe

Yoruba Igbo Hausa/Fulani

Other
 SECTION B: ASSOCIATED RISK FACTORS

Have you donated blood before?

Yes No

Why are you donating blood? Voluntary

Commercial

Family/Relative Replacement

Have you been given blood product?

Yes No

If single, are you sexually active?

Yes No

Number of sexual partners in the last five years

1 2 3 >3

Have you had any sexually transmitted diseases before?

Yes No

Do you use intravenous drugs?

Yes No

If yes, do you share needles with your friends

Yes No

Do you smoke cigarettes?

Yes No

Do you consume alcohol?

Yes No

Do you have history of unprotected casual sex?

Yes No

Yes No

APPENDIX II
 CONSENT FORM

I, ………………………………………………….., hereby give my consent to participate in

the research ‘Serosuveillance Of HIV P-24 Antigens And Antibodies Among Prospective

Blood Donors’ In Ore, Ondo State.

I understand that a blood sample shall be taken from me and that no harm will come to me as

a result of the procedure or the study.

I also understand that, I shall not be victimized in any way if I choose not to participate in this

study.

I have noted the contact details of the researcher, and acknowledge that my privacy shall be

protected.

Sign……………………………..………………..

Date………………………………………………