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Liver Enzyme Profile of Hepatitis B Positive Blood Donors in Enugu

State University Teaching Hospital

Humphrey Nwobodo

, Uzoma Vision Onyinyechi

, Somtochukwu Chukwunweike Ezenwalie

*, Ozioma

Peace Israel

, Lydia Chinenye Madukaife

, Juliet Chinaza Ezeachogu

Enugu State University of Science and Technology, Nigeria

Nnamdi Azikiwe University, Nigeria

DOI: https://doi.org/10.51244/IJRSI.2025.120800353

Received: 05 Sep 2025; Accepted: 12 Sep 2025; Published: 14 October 2025

ABSTRACT

Hepatitis B virus (HBV) remains a major global health problem. This is particularly seen through blood

donation, where many carriers are asymptomatic and can unknowingly transmit the virus. This study addresses

a gap in local data by investigating the liver enzyme profiles of HBV-positive blood donors at the Enugu State

University Teaching Hospital. We conducted a descriptive, cross-sectional study from September to November

2024, enrolling 170 voluntary blood donors. Blood samples were screened for Hepatitis B surface antigen

(HBsAg) using a rapid diagnostic strip, and the liver enzymes—aspartate aminotransferase (AST), alanine

aminotransferase (ALT), and alkaline phosphatase (ALP)—were measured using standard biochemical assays.

Questionnaires were also used to collect socio-demographic data and assess HBV awareness.

The study found a 17.5% HBsAg seroprevalence among the 97 donors included in the final analysis. A high

level of HBV awareness (81.4%) was observed, with most knowledge gained from health workers. The donor

population was predominantly male (89.7%), and the highest prevalence of HBV-positive cases (54.6%) was

in the 26–35 age group. The liver enzyme profiles of the 17 HBsAg-positive donors showed mildly elevated

levels, particularly for ALP. The mean values were: AST 17.82 U/L (range 5.0–38.0), ALP 62.02 U/L (range

22.1–108.1), and ALT 11.12 U/L (range 4.0–26.0). These mild elevations suggest potential subclinical liver

damage, consistent with either a mild acute or chronic viral hepatitis, emphasizing that even asymptomatic

carriers can have subtle liver pathology. The findings highlight the critical need for continued rigorous

screening and further clinical investigation of seropositive donors, including comprehensive follow-up and

advanced testing (e.g., HBV DNA), to ensure transfusion safety and mitigate long-term health risks.

Keywords: Hepatitis B Virus, Liver Enzyme Profile, Blood Donors, Seroprevalence, Transfusion Safety

INTRODUCTION

Hepatitis B is a potentially life-threatening liver infection caused by Hepatitis B virus and it is a major global

health problem (WHO, 2025). The World Health Organization estimates that hundreds of millions of people

are living with chronic HBV, which can lead to severe liver diseases, including cirrhosis and hepatocellular

carcinoma (WHO, 2021). The asymptomatic nature of the infection in many of its early stages is particularly

problematic, as many carriers are unaware of their clinical status and can unknowingly transmit the virus,

making them a potential source of infection to others (Bosch, 2019). This poses a critical challenge in

transfusion medicine, where ensuring a safe blood supply is paramount. Despite rigorous screening protocols

for Hepatitis B surface antigen (HBsAg), asymptomatic carriers with subclinical or even undetectable liver

enzyme changes may still contribute to the transmission of the virus (Hoofnagle, 2012).

Infection with HBV primarily targets hepatocytes, triggering an immune response that, while fighting the

virus, can also cause liver cell damage (WHO, 2021). Over time, this can lead to progressive liver

inflammation and damage, potentially resulting in liver fibrosis, cirrhosis, and cancer (Thad et al., 2019). Liver

enzymes, particularly alanine aminotransferase (ALT) and aspartate aminotransferase (AST), are crucial

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biomarkers of liver cell injury and are often used to monitor liver function (Ganem, 2014). Globally, HBV

infection contributes to approximately 68,600 annual deaths and is a leading cause of liver cancer, with over

300,000 attributed deaths per year (WHO, 2015; W.H.O.E. Region, 2017).

In Nigeria, a country with high endemicity, the seroprevalence of HBV among blood donors ranges from 8%

to 15%, with variations depending on geographical location (Ejele et al., 2021). While a great deal of research

has focused on the prevalence of HBsAg among blood donors, there is a critical knowledge gap concerning the

liver health of these seropositive, yet asymptomatic, individuals. It is ethically and clinically imperative that a

seropositive result prompts further investigation beyond simple discharge from the blood donation center. A

comprehensive assessment, including liver enzyme profiling, is necessary to determine the extent of liver

involvement and to guide appropriate follow-up care.

This study was designed to bridge this gap by investigating the liver enzyme profiles of HBsAg-positive blood

donors at the Enugu State University Teaching Hospital. By providing a detailed analysis of these biomarkers,

this research will offer insights into the subclinical effects of HBV infection on liver function in this key

population. The findings will not only reinforce the need for meticulous screening but will also provide a

robust scientific basis for implementing comprehensive post-donation counseling and long-term management

strategies for seropositive donors. Ultimately, this research aims to enhance the safety of blood transfusions

and contribute to the broader efforts of HBV control and management in Nigeria.

Statement of the Problem

Hepatitis B virus (HBV) infection is the most common cause of liver disease and liver cancer, despite

education and availability of an efficacious vaccine(Tan et al., 2020). Screening for hepatitis B virus infection

is simple and relatively inexpensive. Yet it is underused in everyday practice, leaving some HBV-positive

patients unaware and at risk for serious health consequences, including cirrhosis, liver failure, and

hepatocellular carcinoma (Rizzo et al., 2022). While routine screening for Hepatitis B surface antigen (HBsAg)

is standard practice, there is a lack of data on the liver health status of these asymptomatic, HBsAg-positive

donors, particularly in the Enugu State region of Nigeria.

This study aims to address this critical knowledge gap by determining the liver enzyme profiles of HBsAg-

positive blood donors at the Enugu State University Teaching Hospital. Liver enzymes, such as aspartate

aminotransferase (AST) and alanine aminotransferase (ALT), are key indicators of liver cell damage and

function. By analyzing these biomarkers, we seek to uncover potential subclinical liver pathology in seemingly

healthy donors.

Justification of the Study

Early detection of liver damage, especially in Hepatitis B positive individuals, is critical in preventing the

progression to more severe liver diseases. Blood donation is an essential part of healthcare systems globally,

providing lifesaving treatments for individuals requiring transfusions (Alao et al., 2019). Despite rigorous

screening protocols, asymptomatic carriers of Hepatitis B with mild or undetectable liver enzyme changes may

still contribute to the transmission of the virus (Hoofnagle, 2012).

The findings from this research will not only highlight the importance of rigorous screening but will also

provide a robust scientific basis for implementing comprehensive post-donation follow-up and management

strategies for seropositive donors. Ultimately, this study's contribution is to enhance the safety of blood

transfusions and contribute to the ongoing efforts to control and manage HBV infection in Nigeria.

Significance of the Study

This study will provide valuable insights into the liver health of HBV-positive blood donors in Enugu State

University Teaching Hospital, potentially leading to improved screening protocols and safer blood transfusion

practices. The findings could also guide clinical management of HBV in blood donors and help mitigate the

risks associated with liver damage in these individuals.

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Aim

To Determine Liver Enzyme Profile of Hepatitis B Positive Blood Donors in Enugu State University Teaching

Hospital, G.R.A Enugu.

Specific Objectives:

1. To Screen Blood Donors for Hepatitis B Virus attending Enugu State University Teaching Hospital.

2. To determine their Liver Enzyme Profile Particularly (AST, ALT and ALP) of Hepatitis B Positive Blood

donors in Enugu State University Teaching Hospital.

3. To Asses awareness among blood donors attending Enugu State University Teaching Hospital.

METHODOLOGY

Study Design

A descriptive, cross-sectional study was conducted using a consecutive sampling method, enrolling all eligible

voluntary blood donors who presented at the Blood Bank Centre of Enugu State University Teaching Hospital

from July to September 2024. This a hospital is a major reference hospital for the whole southeastern region of

Nigeria and so was a good representation.

Study Area

The study was carried out in Enugu State University Teaching Hospital (Parklane). G.R.A Enugu Urban, the

capital city of Enugu state, Nigeria.

Study Population

People who Tested Sero-Positive for Hepatitis B virus among Blood donors attending ESUTH, Parklane,

during the study period.

Inclusion Criteria

 Patients Tested Sero-Positive with Hepatitis B virus.

 Individuals who provided informed consent to Participate in this Study.

 All eligible blood donors aged 18-65 years.

 Hemoglobin level within the normal range for male ≥13.5 mg/dl; for female ≥12 mg/dl

Exclusion Criteria

 Donors with known liver disease or other co-infections (HIV, Hepatitis C).

 Blood donors who refused to give their consent.

Sample Size determination

The sample size was calculated using the standard formula for calculating the minimum sample size (Charan J,

2013). Sample size (n) is given by Crochan’s formula

n= Z^2pq

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d^2

Where;

n = Minimum sample size =?

Z = Standard normal deviation at 95% level of confidence = 1.96

P = (the percentage of target population estimated to have a particular characteristic) = 50% (0.5)

q = 1-p = 1-0.5 = 0.5

d (margin of error) = 10% (0.1)

Therefore, n = (1.96^2 * 0.5 * 0.5)

0.1^2

Sample size = 97.

The sample size targeted at 97 subjects.

Sample Collection

The blood sample was taken from each participant aseptically for the serological with a sterile disposable

syringe and needle, after disinfection of the selected venipuncture site with 70% alcohol in an expanding

circular scrub from the center to the periphery of the needle insertion. About 5ml of blood was collected by

venipuncture and was dropped into a plain bottle labeled with corresponding sample number, the plasma was

separated and used for serology For Hepatitis b virus screening.

Laboratory procedure

Serum was separated from the collected blood samples in a test tube by centrifuging at 2000 rpm for 2

minutes. The separated blood samples were serologically investigated for viral infection of hepatitis B by

Palmitic Hepatitis B Rapid Diagnostic strip. All blood collection was done following laboratory protocols

ensuring the accuracy and reliability of the biochemical analysis. The 17 hepatitis B positive (HBsAg positive)

serum samples were studied for liver function tests, estimation of levels of alanine aminotransferase (ALT),

aspartate aminotransferase (AST) and alkaline phosphatase (ALP) by using test kits (RANDOX company, UK

and Human Germany).

Serology (Hepatitis B Screening) Materials

Sterile needle, HBsAg rapid test strip, Sample (whole blood, serum, or plasma, Buffer solution (if required),

Dropper or pipette.

Interpretations of Result

Positive: Two lines appear, one line should always appear in the control line region (C) and another one

apparent colored line should appear in the test line region.

Negative: Once colored line appears in the control region (C). No apparent colored line appear in the test line

region.

Invalid: Control line fails to appear; insufficient specimen volume or incorrect procedural techniques are the

most likely reasons for control line failure. Review the procedure and repeat the test with a new strip. If the

problem persists, discontinue using the test kit immediately and contact your local distributor.

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Alanine Transaminase

Method and Materials

Spectrophotometry (Reitman-Frankel). Buffer/Substrate, 2-4 dinitrophenylhydrazine, water (distilled water),

0.4N NaOH (Sodium Hydroxide solution), Water bath at 37℃, test tube and control serum. The buffer is

called Tris’s buffer and is contained with substrate which is 2-oxoglutarate.

Aspartate Transaminase.

Method and Material

Spectrophotometry was used. Buffer/Substrate, 2-4 dinitrophenylhydrazine, water (distilled water), 0.4N

NaOH (Sodium Hydroxide solution), Water bath at 37℃, test tube and control serum. The buffer is called

Tris’s buffer and is contained with substrate which is alpha ketoglutarate and is measured at 540nm.

Alanine Phosphatase.

Method & Materials

Spectrophotometry (according the standardized method described by DGKC). Buffer/Substrate, 2-4

dinitrophenylhydrazine, water (distilled water), 0.4N NaOH (Sodium Hydroxide solution), Water bath at 37℃,

test tube and control serum. ALP is measured at 405nm.

Procedure

 Into a cuvette was added 0.5 ml of working reagent and 0.01 ml of sample, it was mixed and initial

absorbance read at 405 nm and a stop watch started

 The absorbance was further read at 1, 2, and 3 minutes respectively.

 The change in absorbance was then multiplied with factor 2760 to get the activity of the alkaline

phosphatase enzyme.

Data Collection and Analysis.

The socio-demographic data and other relevant information of each participant were obtained using a self-

administered questionnaire. Data obtained from this study was analyzed using the statistical package for social

sciences (SPSS). Data was presented as mean and standard deviations

Ethical consideration

Ethical clearance for the study was sought and obtained from the ethical committee of Enugu State University

Teaching Hospital (ESUTH), Parklane, with the reference number, ESUTH/HREC/2024/10/201.

RESULTS

Table 1: Sociodemographic Data (n = 97)

Variables

Frequency

Percentage (%)

Gender

- Male

89.7

-Female

10.3

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Age

-18–25

22.7

-26–35

54.6

-36–45

17.5

-46 and above

5.2

Marital Status

-Single

59.2

-Married

36.7

-Divorced

4.1

-Widowed

Level of Education

- No formal education

10.3

- Primary school

17.5

- Secondary school

43.3

- Tertiary Education

28.9

- Occupation

- Student

26.8

- Employed

17.5

- Self-employed

36.1

- Unemployed

19.6

Have you donated blood before?

- Yes

82.5

- No

17.5

Table 1 presents the demographic characteristics of the respondents. The study's participants were

predominantly male (89.7%), with the largest age group being 26–35 years old (54.6%). The majority were

single (59.2%) and had completed secondary school (43.3%). In terms of occupation, most were self-employed

(36.1%), followed by students (26.8%). A significant majority of participants (82.5%) reported a history of

previous blood donation.

Table 2: Awareness of Hepatitis B Virus (n = 97)

Variables

Frequency

Percentage (%)

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Have you ever heard of Hepatitis B Virus (HBV)

- Yes

81.4

- No

18.6

If yes, where did you learn about HBV?

- Health workers

72.4

- Media (TV, radio, internet)

12.6

- Friends/Relatives

5.6

Do you know that HBV is a serious liver infection?

- Yes

38.1

- No

61.9

What do you think are the main ways HBV is transmitted?

- Blood transfusion

23.2

- Unprotected sexual intercourse

20.7

- Sharing needles/syringes

20.7

- Mother-to-child during birth

15.6

-Contact with contaminated items (e.g., razors, toothbrushes)

19.8

Are you aware that HBV can be prevented through vaccination?

- Yes

78.2

- No

21.8

Do you believe screening for HBV before blood donation is important?

- Yes

100

- No

Table 2 presents the awareness of Hepatitis B Virus (HBV) among respondents. Awareness of HBV was high,

with 81.4% of respondents having heard of the virus. The primary source of information was health workers

(72.4%). While 38.1% of participants considered HBV a serious infection, all respondents (100%) supported

pre-donation screening. The most recognized transmission routes were blood transfusion, unprotected sexual

intercourse, and sharing needles.

Table 3: Prevalence of Hepatitis B among Blood donors at ESUTH Parklane.

Variables

Prevalence

Blood donors screened

100.0

Hepatitis B positive donors

17.5

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Table 3 shows the prevalence of Hepatitis B among blood donors at ESUTH Parklane. Out of 97 blood donors

screened, 17.5% tested positive for Hepatitis B, indicating a moderate level of the infection within the donor

population.

Table 4: Liver Enzyme Profile of Hepatitis B Positive Blood donors

Minimum

Maximum

Mean

Std. Deviation

AST

5.0

38.0

17.82

10.80

ALP

22.1

108.1

62.02

27.56

ALT

4.0

26.0

11.12

6.13

Table 4 shows the Liver Enzyme Profile of Hepatitis B Positive Blood donors. For aspartate aminotransferase

(AST) showing a narrower range, values range from a minimum of 5.0 to a maximum of 38.0 with a mean of

17.82 and a standard deviation of 10.80. Alkaline phosphatase (ALP) measurements shows a significant range,

from a minimum of 22.1 to a maximum of 108.1, with a mean of 62.02 and a standard deviation of 27.56.

Alanine aminotransferase (ALT) levels, with a mean of 11.12 and a smaller standard deviation of 6.13, range

from 4.0 to 26.0.

Figure 1: Prevalence of Hepatitis B Positive Cases Among Donors by Gender

Figure 1 illustrates the prevalence of Hepatitis B among male and female blood donors. The prevalence was

higher in male donors (82.4%) than in female donors (17.6%).

Figure 2: Prevalence of Hepatitis B Positive Cases Among Donors by Age

82.40%

17.60%

Male

Female

18–25

26–35

36–45

46 and above

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This chart illustrates the prevalence of Hepatitis B positive cases among different age groups of blood donors.

The 26–35 age group had the highest frequency, with 6 positive cases. The 36–45 age group followed with 4

positive cases. The 18–25 and 46 and above age groups each had 4 and 3 positive cases, respectively.

Figure 3: Prevalence of Hepatitis B Positive Cases Among Donors based on Marital Status

This chart shows the Prevalence of Hepatitis B positive cases based on the marital status of blood donors. The

majority of positive cases were found among single donors, with a frequency of 10 positive cases. Married

donors had a lower frequency, with 7 positive cases. There were no positive cases reported among divorced

donors.

Figure 4: Prevalence of Hepatitis B Positive Cases Among Donors based on Awareness of Hepatitis B

This chart displays the prevalence of Hepatitis B positive cases based on whether blood donors have heard of

Hepatitis B Virus (HBV). Among donors who have heard of HBV, 11 individuals tested positive for Hepatitis

B. In contrast, among those who have not heard of HBV, 6 individuals tested positive.

DISCUSSION

The result of this study showed that A moderate HBsAg seroprevalence of 10% was observed among the 170

screened blood donors, with 17 individuals testing positive for HBV. This prevalence is considered moderate

when compared to other regional findings, such as an 18.1% prevalence reported in Maiduguri, Borno State,

Nigeria (Seto et al., 2014). The study's discussion suggests that this comparatively lower prevalence might be

attributed to the high level of HBV awareness among the donor population. Indeed, a significant 81.4% of

Single Married Divorced

Yes No

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respondents reported having heard of HBV, primarily through health workers (72.4%). While awareness of the

virus was high, only 38.1% of participants acknowledged HBV as a serious liver infection. Nevertheless, all

respondents (100%) emphasized the importance of screening for HBV before blood donation, highlighting a

strong public understanding of its role in transfusion safety. The most recognized transmission routes included

blood transfusion, unprotected sexual intercourse, and sharing needles/syringes. This underscores the critical

importance of rigorous screening to prevent the transmission of this potentially life-threatening virus through

blood donations, as asymptomatic carriers can unknowingly transmit the virus.

Demographic analysis of the study participants revealed a strong male dominance (89.7%) in the donor

population. This finding contradicts some previous reports (Uneke, 2005) but is consistent with others that

found higher blood donation rates among males in both rural and urban areas, including studies in Lagos,

Nigeria (Balogun, 2010), and Ibadan, Nigeria (Lukhwareni, 2009; Mehmet, 2005). The highest prevalence of

HBV-positive cases (54.6%) was observed in the 26–35 age group. This aligns with studies noting higher HBV

prevalence in younger individuals aged 20–29 years (Buseri, 2009), possibly due to more active sexual

activities and other risk behaviors. This finding contrasts with earlier reports suggesting a higher prevalence in

older subjects (Lawal, 2009; Luka, 2008). The majority of HBV-positive cases were found among single

donors, with 10 cases, compared to 7 among married donors. Most respondents had completed secondary

education (43.3%) and were self-employed (36.1%). A significant majority (82.5%) reported a history of

previous blood donation.

The liver enzyme profiles of the 17 HBsAg-positive donors showed mild elevations in Aspartate

aminotransferase (AST), Alanine aminotransferase (ALT), and Alkaline phosphatase (ALP). Specifically, AST

ranged from 5.0-38.0 U/L (mean 17.82 U/L), ALP from 22.1-108.1 U/L (mean 62.02 U/L), and ALT from 4.0-

26.0 U/L (mean 11.12 U/L). These values suggest subtle liver damage, consistent with either a mild acute or

chronic viral hepatitis.

It is important to note that AST is not a specific marker solely for liver damage, as its mild elevation can be

attributed to various factors, including intense exercise. Given AST's shorter half-life of 17 hours, a slight

elevation might indicate an "acute phase" of viral infection. In contrast, ALT is considered a more specific

marker for liver damage, and its longer half-life (48 hours) often associates elevations with chronic liver

conditions. The relatively lesser elevations in both ALT and AST compared to ALP in this study, while still

suggestive of viral hepatitis, indicate either mild acute viral hepatitis or diffuse and focal chronic liver diseases.

Elevated ALP levels typically reflect impaired biliary tract function and could be indicative of a recent

hepatitis B attack. Although liver function test results can be normal in HBsAg carriers, severe cases of liver

disease like cirrhosis or fulminant liver failure commonly show AST and ALT values exceeding 1000 U/L.

The observed mild increases, particularly in ALP, are crucial in suggesting subclinical liver damage, even in

asymptomatic carriers.

CONCLUSION

The study confirms that HBV infection significantly affects liver enzyme profiles, leading to subtle liver

damage. The findings shows the critical importance of rigorous screening, including routine liver function tests

for HBV seropositive individuals, and the necessity for further clinical investigation of seropositive donors.

These measures are vital to mitigate long-term health impacts on individuals and enhance transfusion safety by

preventing the spread of HBV through blood donations

RECOMMENDATION

1. It is recommended that all blood donors undergo comprehensive HBV screening, including HBsAg and,

where feasible, additional markers or HBV DNA testing, as part of the routine blood donation process,

particularly during their first visit, to ensure maximum transfusion safety

2. HBV seropositive individuals should undergo regular and comprehensive clinical follow-up, including

routine liver function tests to monitor enzyme levels and detect liver damage early. This follow-up should

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ideally include HBV DNA testing and other serological markers to assess disease activity and stage (acute vs.

chronic), guiding appropriate clinical management and mitigating long-term health impacts.

3. Increased public health campaigns are needed to educate individuals about the Causes of HBV, their

transmission route, prevention and treatment.

ACKNOWLEDGEMENTS

The Dean of faculty of Medical Laboratory Science and the head of the laboratory for approval of the use of

the faculty laboratory facility; all participants for volunteering to be part of the study.

Competing Interests

Authors have declared that no competing interests exist.

Consent

All authors declare that written informed consent was obtained from the participants for publication of this

case report and accompanying images. A copy of the written consent is available for review by the Editorial

office/Chief Editor/Editorial Board members of this journal.

Ethical Approval

All authors hereby declare that all experiments have been examined and approved by the appropriate ethics

committee and have therefore been performed in accordance with the ethical standards laid down in the 1964

Declaration of Helsinki.

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