Viral immunochemical status of HBeAg, HBeAb, Anti-HIV, and Anti-HCV in hepatitis B surface antigen-seronegative anicteric malaise patients

Mathew Folaranmi Olaniyan; Opeoluwa Oluwabamise Jegede

doi:10.4103/bbrj.bbrj_120_18

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ORIGINAL ARTICLE

Year : 2018 | Volume : 2 | Issue : 4 | Page : 286-289

Viral immunochemical status of HBeAg, HBeAb, Anti-HIV, and Anti-HCV in hepatitis B surface antigen-seronegative anicteric malaise patients

Mathew Folaranmi Olaniyan¹, Opeoluwa Oluwabamise Jegede²
¹ Department of Medical Laboratory Science, Edo University, Iyamho, Nigeria
² Department of Medical Laboratory Science, Achievers University, Owo, Nigeria

Date of Submission	10-Sep-2018
Date of Decision	15-Oct-2018
Date of Acceptance	20-Oct-2018
Date of Web Publication	11-Dec-2018

Correspondence Address:
Dr. Mathew Folaranmi Olaniyan
Department of Medical Laboratory Science, Edo University, Iyamho
Nigeria

Source of Support: None, Conflict of Interest: None

DOI: 10.4103/bbrj.bbrj_120_18

Abstract

Introduction: Malaise involves general discomfort, uneasiness, or pain which could indicate microbial infection or other diseases which could be as a result of immune response. Methods: The study population was 200 malaise anicteric hepatitis B surface antigen (HBsAg)-seronegative patients (females – 100; males – 100) aged 5–76, and 111 apparently healthy anicteric nonmalaise age-matched subjects who were HBsAg-seronegative (females – 56 and males – 55) were recruited as test and control subjects, respectively. Immunochemical assay of anti-HBe, anti-HCV, HBsAg, and HBeAg was carried out in all the participants by enzyme-linked immunosorbent assay, while anti-HIV was determined in the participants by immunochromatographic and Western blot assay. Results: The results obtained in HBsAg-seronegative anicteric malaise patients revealed the frequency of occurrence of 3% (6) HBeAg; 9.5% (19) HBeAb; 3.5% (7) anti-HIV; 6.5% (13) anti-HCV; 1% (2) anti-HIV + HBeAg; 0.5% (1) anti-HIV + HBeAb; 0.5% (1) anti-HIV + anti-HCV; and 0.5% (1) anti-HIV + anti-HCV + HBeAg, with 9.5% (19) HBeAb found to be most frequent followed by 6.5% (13) anti-HCV; 3.5% (7) anti-HIV and 3% (6) HBeAg were more in female than male. About 25% (50/200) malaise HBsAg-seronegative patients expressed at least one of the HIV, HCV, and HBV serologic markers compared with 9.9% (11/111) in the control subjects. There was a low frequency of coexpression of viral biomarkers (0.5%–1%). Conclusion: Malaise in HBsAg-seronegative patients could be associated with the expression of serologic biomarkers of HCV, HIV, and HBV.

Keywords: Anicteric, hepatitis B surface antigen seronegative, malaise, viral immunochemical biomarkers

How to cite this article:
Olaniyan MF, Jegede OO. Viral immunochemical status of HBeAg, HBeAb, Anti-HIV, and Anti-HCV in hepatitis B surface antigen-seronegative anicteric malaise patients. Biomed Biotechnol Res J 2018;2:286-9

How to cite this URL:
Olaniyan MF, Jegede OO. Viral immunochemical status of HBeAg, HBeAb, Anti-HIV, and Anti-HCV in hepatitis B surface antigen-seronegative anicteric malaise patients. Biomed Biotechnol Res J [serial online] 2018 [cited 2023 Jun 10];2:286-9. Available from: https://www.bmbtrj.org/text.asp?2018/2/4/286/247242

Introduction

Malaise occurs when an individual has a feeling of general discomfort, fatigue, and pain which may be an indication of infection.^[1]

Individuals who do not possess antibody to HBc, HBs, HBe, HBe antigen, and HBs antigen are not infected with HBV but are susceptible to the infection. Those who are immune due to natural infection will not express HBs antigen or HBe antigen but will possess antibody to HBc, HBe, and HBs. Individuals who have immunity to hepatitis B as a result of vaccination will not express of anti-HBe, HBeAg, anti-HBc, and hepatitis B surface antigen (HBsAg) but anti-HBs in their serum. The following is expressed in active infection: HBsAg, anti-HBc, anti-HBe, HBeAg, immunoglobulin M (IgM) anti-HBc, and anti-HBs.^[2]

Infection of hepatitis B virusis associated with malaise that starts with general ill health, loss of appetite, nausea, vomiting, body aches, mild fever, itchy skin, dark urine, and jaundice.^[3]

Expression of antibodies HCV and HIV indicates HCV and HIV infection, respectively.^[4],[5],[6]

Signs of viral infection include fatigue, loss of appetite, weight loss, fevers, night sweats, chills, aches, viral conjunctivitis or “pink eye,” itchy, burning sensation, jaundice, and pains.^[7]

An individual could be HBsAg seronegative due to clearance of the antigen from the body or when such an individual is not infected. Malaise is a nonspecific sign/symptom which includes general discomfort and pain which used to be the first indication of an infection or disease. There is little information on the frequency of HBeAg, HBeAb, anti-HIV, and anti-HCV in HBsAg-seronegative anicteric malaise patients. This work determined the pattern of HBeAg, HBeAb, anti-HIV, and anti-HCV in HBsAg-seronegative anicteric malaise patients.

Methods

Study area

Ado Ekiti is the state capital of Ekiti state in South Western zone of Nigeria. It hosts Ekiti State University Ado-Ekiti, Afe Babalola University, Federal Polytechnic, Ado-Ekiti, Crown polytechnic, Nigerian Television Authority, Ekiti State Television, Radio Ekiti, and Progress FM and Voice FM radio stations. The city is a major trade center for yams, cassava, grain, cotton, and tobacco.

Study population

The study population included 200 malaise anicteric HBsAg-seronegative patients (females – 100 and males – 100) aged 5–76 years attending Clinical Outpatient of Ekiti State Teaching Hospital, Ado-Ekiti, Nigeria, and 111 apparently healthy nonmalaise age-matched subjects who were HBsAg seronegative (females – 56 and males – 55) were recruited as test and control subjects, respectively.

Immunochemical assays

Determination of hepatitis B viral biomarkers:

Patients were pre- and posttest counseled for the tests
HBsAg test was carried out in all the volunteers using the reagent kit of BIO–RAD Raymond Poincare, Marnes-la Coquette.

Principle

This is a sandwich enzyme-linked immunosorbent assay that uses three monoclonal antibodies that could bind to their corresponding subtypes of HBsAg. The solid phase is polystyrene microwells coated with the first monoclonal antibody, while the other two monoclonal antibodies are bound to the enzyme peroxidase. They bind their corresponding HBsAg to generate an observable reaction in form of color formation.

The assay procedure includes the following reaction steps:

Distribution of samples into the wells of the microplates: This distribution can be visually controlled; there is a clear difference of coloration between the empty well and well with the sample. This distribution can also be controlled automatically by reading at 450/620–700 nm (optional)
Distribution of the conjugate into the wells: This distribution can also be visually controlled; the conjugate which is initially orange becomes red after addition into the well. It is possible to control automatically this distribution by spectrophotometric reading at 450/620–700 nm (optional). The sample deposition can also be controlled at this step of the manipulation by automatic reading at 450/620–700 nm
Incubation at 37°C
Washing and development of the enzyme activity-bound solid phase by the addition of substrate
Stopping development, then reading of the optical densities at 450/620–700 nm and interpretation of the results
HBeAg and anti-HBe tests were determined in the plasma of the subjects using the reagent kit of DIA. PRO, Diagnostic Bioprobes Srl Via Columella, Milano, Italy.

Principle of the test

HBeAg: Hepatitis B envelope antigen (HBeAg) in the plasma binds with its corresponding anti-HBeAg antibodies after incubation and washing this complex in turn binds the peroxidase enzyme added to the microplate. The reaction is made observable by the addition of chromogen substrate
Anti-HBeAg: anti-Hbe in the plasma or serum competes with recombinant HBeAg for the binding site on anti-HBeAg antibody on the microplate wells. Two anti-HBeAg monoclonal antibodies bind to peroxidase and chromogen–substrate complex is added for the formation of color.

HIV tests

HIV screening was carried out on the participants using Abbott Determine HIV-1/2, a reagent kit of Abbot Laboratories Co., Ltd., Japan.

HIV confirmatory test

The HIV confirmatory test was carried out on all the participants by Western blot assay, using reagent kit of Immunoetics, Inc., 27 Dryclock Avenue, Boston, USA. http//www.immunoetics.com

Anti-HCV test

Anti-HCV test was determined on all the participants using the reagent kit of DIA.PRO, Diagnostic Bioprobes Srl Via Columella, Milano, Italy.

Principles of the test microplates are attached with HCV-specific antigens which bind with the antibody from the plasma or serum. The reaction is made observable by the addition of antiglobulin and IgM antibody, labeled with peroxidase.

Ethical consideration

The proposal was reviewed and approved by the Research and Ethical committee of Ekiti state Teaching Hospital, Ado-Ekiti, Nigeria, before the commencement of the work. Consent of each of the participants was also obtained. The Ethics Committee Approval number was Ado-Ekiti, Nigeria. 2017 and date was 2016 -2018.

Data analysis

The results obtained were subjected to statistical analysis to evaluate the mean, frequency, and percentage using the Statistical Package for the Social Sciences version 18.0.

Results

The results obtained showed a frequency of occurrence of 3% (6) HBeAg; 9.5% (19) HBeAb; 3.5% (7) anti-HIV; 6.5% (13) anti-HCV; 1% (2) anti-HIV + HBeAg; 0.5% (1) anti-HIV + HBeAb; 0.5% (1) anti-HIV + anti-HCV; and 0.5% (1) Anti-HIV + Anti-HCV + HBeAg [Table 1] and [Figure 1].

Table 1: Pattern of immunochemical viral seromarkers obtained in the participants

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Figure 1: The frequency of HIV, HBV, and HCV seromarkers obtained in the participants

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About 9.5% (19) HBeAb was most frequent, followed by 6.5% (13) anti-HCV; 3.5% (7) anti-HIV and 3% (6) HBeAg were more in females than males. There was a low frequency of coexpression of viral biomarkers: 1% (2) anti-HIV + HBeAg; 0.5% (1) anti-HIV + HBeAb; 0.5% (1) anti-HIV + anti-HCV; and 0.5% (1) anti-HIV + anti-HCV + HBeAg [Table 1] and [Figure 1].

About 25% (50/200) malaise HBsAg-seronegative patients expressed at least one of the HIV, HCV, and HBV serologic markers compared with 9.9% (11/111) in the control subjects [Table 1].

Discussion

This study was used to determine the pattern of HBe antigen, HBe antibody, HIV antibody, and HCV antibody in HBsAg-seronegative anicteric malaise patients. The results obtained showed a frequency of immunochemical viral biomarkers as follows: 3% (6) HBeAg; 9.5% (19) HBeAb; 3.5% (7) anti-HIV; 6.5% (13) anti-HCV; 1% (2) anti-HIV + HBeAg; 0.5% (1) anti-HIV + HBeAb; 0.5% (1) anti-HIV + anti-HCV; and 0.5% (1) anti-HIV + anti-HCV + HBeAg. Approximately 9.5% (19)HBeAb was most frequent, followed by 6.5% (13) anti-HCV; 3.5% (7) anti-HIV and 3% (6) HBeAg were more in females than the males. HBeAb indicates clearance of the envelope antigen to hepatitis B virus and convalescence though the patient is hepatitis B virus infected. HBeAg indicates active hepatitis B virus infection and replication. The presence of antibodies to HIV and HCV is an indication of HIV and HCV infection, respectively. Antibody to HBe which is most prevalent in this work could be attributed to high prevalence of Hepatitis B virus relative to the prevalence of HIV and HCV in Nigeria^[8] which Otegbayo et al.^[9] reported as 11.9%, HBsAg and 4.8%, anti-HCV. Otegbayo et al.^[9] also reported gender difference in HCV and HBV serologic markers. HBsAg was more common among males than females, while anti-HCV was found in a similar proportion in males and females.

There was a low frequency of coexpression of viral biomarkers: 1% (2) nti-HIV + HBeAg; 0.5% (1) anti-HIV + HBeAb; 0.5% (1) nti-HIV + nti-HCV; and 0.5% (1) nti-HIV + nti-HCV + HBeAg. This finding agrees with the report of Muriuki et al.^[10] on the prevalence of hepatitis B and C viral coinfections among HIV-1-infected individuals in Nairobi, Kenya. Muriuki et al.^[10] reported low levels of coinfection with all three viruses, but there could be higher prevalence rates specifically in high-risk populations. The low frequency of viral coinfections found in this study could also be higher but possibly due to high-death rate associated with viral coinfections as HIV-positive individuals coinfected with HBV and/or HCV have increased rates of progression to chronic hepatitis infection, fibrosis, cirrhosis, hepatocellular carcinoma, and end-stage liver disease.^{[1],[11],[12],[13],[14]} About 25% (50/200) malaise HBsAg-seronegative patients expressed at least one of the HIV, HCV, and HBV serologic markers compared with 9.9% (11/111) in the control subjects. This finding could be explained as malaise involves general discomfort, uneasiness, or pain, which is the first thought as an indication of infection or other diseases as signs and symptoms of viral infection of the viruses in this work include fatigue, nausea, muscle or joint pains, general ill-health, loss of appetite, nausea, vomiting, body aches, and mild fever.^[3],[6],[15]

Conclusion

The work also showed a frequency of occurrence of 3% (6) HBeAg; 9.5% (19) HBeAb; 3.5% (7) anti-HIV; 6.5% (13) anti-HCV; 1% (2) anti-HIV + HBeAg; 0.5% (1) anti-HIV + HBeAb; 0.5% (1) anti-HIV + anti-HCV; and 0.5% (1) anti-HIV + anti-HCV + HBeAg, with 9.5% (19) HBeAb found to be most frequent followed by 6.5% (13) anti-HCV; 3.5% (7) anti-HIV and 3% (6) HBeAg were more in females than the males. About 25% (50/200) malaise HBsAg-seronegative patients expressed at least one of the HIV, HCV, and HBV serologic markers compared with 9.9% (11/111) in the control subjects. There was a low frequency of coexpression of viral biomarkers (0.5%–1%).

Routine evaluation of HBeAg, HBeAb, anti-HIV, and anti-HCV in HBsAg-seronegative anicteric malaise patients will provide useful direction for effective management of malaise patients.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

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Figures

[Figure 1]

Tables

[Table 1]

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