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 Table of Contents  
ORIGINAL ARTICLE
Year : 2019  |  Volume : 4  |  Issue : 3  |  Page : 81-86

Possible viral immunochemical status of children with elevated blood fibrinogen in some herbal homes and hospitals in Nigeria


Department of Medical Laboratory Science, Edo University, Iyamho, Edo State, Nigeria

Date of Submission21-May-2019
Date of Acceptance16-Sep-2019
Date of Web Publication27-Sep-2019

Correspondence Address:
Mathew Folaranmi Olaniyan
Department of Medical Laboratory Science, Edo University, Iyamho, Edo State
Nigeria
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ed.ed_20_19

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  Abstract 


Study Background: Viral infection in children can elicit acute-phase response which can cause significant alterations in the level of acute-phase proteins such as fibrinogen.
Aim and Objective: This work was designed to determine the possible viral immunochemical status of children aged 3–7 years with elevated blood fibrinogen of >6.0 g/L who received treatments as patients in herbal homes and hospitals in Nigeria.
Materials and Methods: Children with elevated blood fibrinogen >6.0 g/L were recruited from 10 herbal homes (n = 27; 3–7 years) and three hospitals (n = 27; 3–7 years) and children with normal blood fibrinogen (3.1 ± 1.0 g/L; n = 30; 3–7 years) were also studied. Anti-hepatitis C virus (HCV), hepatitis B surface antigen, and human immunodeficiency virus type 1 (HIV1) p24 antigen were determined in each of the children immunochemically by ELISA, while blood fibrinogen was assayed using the Clauss method. Acid-fast bacilli were determined in the sputum by the Ziehl–Neelsen stain, and Plasmodium spp. identification was carried out using Giemsa staining -thick blood film technique.
Results: The viral immunochemical status obtained in children with elevated blood fibrinogen who received treatments in herbal homes showed 3.7% (1) HIV mono-infection; 7.4% (2) HCV mono-infection; 18.5% (5) hepatitis B virus (HBV) mono-infection; 3.7% (1) HIV-HBV coinfection; and 7.4% (2) HCV-HBV coinfection with no coinfection of HIV-HCV and HIV-HCV-HBV, while those who received treatments in the hospitals showed only 11.1% (3) HBV mono-infection. The viral immunochemical status obtained in children with normal blood fibrinogen showed 3.3% (1) HBV mono-infection and 3.3% (1) HCV mono-infection with no HIV mono-infection and HIV-HBV, HIV-HCV, HCV-HBV, and HIV-HCV-HBV coinfections.
Conclusion: Viral seromarkers of HCV, HBV, HIV, HIV-HBV, and HCV-HBV were more in children with elevated blood fibrinogen who received treatments in herbal homes than those who received treatments in the hospital and those with normal blood fibrinogen, which suggests blood fibrinogen as a possible diagnostic indicator in viral infection in herbal homes.

Keywords: Children aged 3–7 years, elevated fibrinogen, herbal homes, hospital, viral immunochemical status


How to cite this article:
Olaniyan MF, Uwaifo F, Ojediran TB. Possible viral immunochemical status of children with elevated blood fibrinogen in some herbal homes and hospitals in Nigeria. Environ Dis 2019;4:81-6

How to cite this URL:
Olaniyan MF, Uwaifo F, Ojediran TB. Possible viral immunochemical status of children with elevated blood fibrinogen in some herbal homes and hospitals in Nigeria. Environ Dis [serial online] 2019 [cited 2022 Nov 29];4:81-6. Available from: http://www.environmentmed.org/text.asp?2019/4/3/81/268149




  Introduction Top


Fibrinogen is a glycoprotein known as blood coagulation factor 1 that circulates in the blood of human being. Tissue and vascular injury can cause enzymatic conversion fibrinogen to fibrin blood clot by thrombin.[1],[2],[3] The primary function of fibrinogen is to block blood vessels to stop excessive bleeding. Fibrin, the product of fibrinogen, can bind and act as antithrombin I to limit blood clotting by reducing the activity of thrombin. Fibrinogen is a positive acute-phase protein which blood levels increase in systemic inflammation, tissue injury, and certain other events.[4],[5],[6],[7]

Infectious viral agents such as human immunodeficiency virus (HIV) and hepatoviruses can trigger systemic inflammation, leading to acute-phase response. Increased acute-phase proteins from the liver may promote sepsis.[5],[7]

A virus is a small infectious particle that replicates only inside the living cells and tissues. Viruses can infect animals, plants, bacteria, and Archaea. Viral infections in human can elicit an immune response to eliminate the infecting viral agent. Immune responses can also be caused by vaccines that provide artificially acquired immunity to the specific viral agent. Viral particles such as HIV and hepatoviruses can evade innate immune responses to cause chronic infections.[8],[9],[10] Hepatitis B virus (HBV) is an hepatotrophic virus that causes inflammation of liver. HBV is a DNA virus transmitted through contact with infectious blood or body fluids. Infection around the time of birth or from contact with other people's blood during childhood is the most frequent method by which hepatitis B is acquired in areas where the disease is common.[11]

Hepatitis C virus (HCV) is an RNA infectious agent that infects and replicates in the liver. It spreads primarily by blood-to-blood contact, poorly sterilized medical equipment, needle stick injuries, transfusions, and from an infected mother to her baby during birth.[12]

HIV, an RNA viral agent, causes HIV infection which may lead to acquired immunodeficiency syndrome. It can elicit cell-mediated immunity.[13],[14] It is a sexually transmitted infection through contact with or transfer of blood, preejaculate, semen, and vaginal fluids. HIV transmission can occur from an infected mother to her infant during pregnancy and during childbirth through contact with her blood or vaginal fluid and breast milk. HIV infects all organs and immune system.[15],[16],[17]

The coagulation and fibrinolysis physiology of newborn and young children differ from older children and adults as a result of the immaturity of the hemostasis system of a newborn, are functionally balanced. Healthy newborns exhibit fewer symptoms of bleeding disorders and thrombosis.[18],[19]

This work was, therefore, designed to determine the possible viral immunochemical status of children aged 3–7 years with elevated blood fibrinogen of >6.0 g/L who received treatments as patients in herbal homes and hospital in Nigeria.


  Materials and Methods Top


Study area

The study area was Saki-West local government area located at the northern part of Oyo state in Nigeria. It shares boundary with Kwara state in Nigeria and Burkina Faso. It hosts a resettlement center of the Second Mechanized Division of Nigerian Army, The Oke-Ogun Polytechnic, Baptist Medical Centre, Muslim Hospital, Baptist School of Nursing, School of Medical Laboratory Technology, Oyo State Hospital, and a Technical College. Saki, Nigeria, is also one of the largest cities in Oyo state.

Study population

Children with elevated blood fibrinogen >6.0 g/L were recruited as test subjects from 10 herbal homes (n = 27; 3–7 years) and three major hospitals (two faith based and a state hospital) (n = 27; 3–7 years), while children with normal blood fibrinogen (3.1 ± 1.0 g/L; n = 30; 3–7 years) in Saki-West, Nigeria, not infected with Plasmodium and who were acid-fast bacilli (AFB)-Ziehl–Neelsen negative were studied as control subject.

Fibrinogen level of children who received treatment in herbal homes and the hospital was determined and those with elevated blood fibrinogen >6.0 g/L were recruited while children with normal blood fibrinogen of 3.1 ± 1.0 g/L were also recruited as controls. All participants who tested negative to Plasmodium infection and AFB-Ziehl–Neelsen staining were included in the study. Samples were taken at the completion of the treatment.

Methods of assay

Fibrinogen measurement using RANDOX kit

Fibrinogen, the most abundant clotting factor in human plasma, is synthesized in the liver. Fibrinogen was analyzed using Randox kit by Clauss method. A high concentration of thrombin is added to dilute test plasma, and the clotting time is measured. The test result was compared with a standard calibration curve.

Identification of Plasmodium spp.

This was carried out by Giemsa-thick blood film technique as described by Cheesbrough.[20]

Giemsa stain is a mixture of azur, methylene blue, and eosin dye. A thick film was prepared. The thick film was air-dried. The Giemsa stain was diluted (5%). The diluted Giemsa stain was allowed to react with the thick blood film on a slide for 30 min and then washed gently in tap water and air-dried. This was examined microscopically using immersion oil objective. The cytoplasm of the parasite is stained with methylene blue while the chromatin dot retains the color of eosin.

Identification of acid-fast bacilli

This was carried out by Ziehl–Neelsen staining technique as described by Cheesbrough.[20]

AFB in a smear of sputum were flooded with strong carbol fuchsin and heated until steam rises to break the waxy cell wall (high lipid cell wall) of the organism to permit the stain. The preparation was decolorized using 3% acid alcohol to differentiate and remove excess stain. This was followed by the addition of malachite green. The stained smear was air-dried, and the back of the slide was cleaned and examined using oil immersion objective. The organism appears as a red slender rod against a green background.

Human immunodeficiency virus 1 p24 ELISA

This was carried out using ABCAM kit. The manufacturer's instructions were strictly adhere to.

On the microtiter tube is attached an affinity tag-labeled antibody and a conjugated detector antibody which binds with HIV1p24 in the sample, standard, and control. The mixture was incubated and washed to remove unbound material. 3,3′, 5, 5′-Tetramethylbenzidine (TMB) substrate was added, and during incubation, this is catalyzed by horseradish peroxidase (HRP) to form blue color. The reaction is stopped by adding stop solution completing any color change from blue to yellow. The intensity is measured at 450 nm.

Hepatitis B surface antigen ELISA assay

This was carried out using Diagnostic Automation/Cortez Diagnostics Inc., Kit. The manufacturer's instructions were strictly adhere to.

The microtiter plate has polystyrene wells coated with mouse monoclonal antibodies specific for hepatitis B surface antigen (HBsAg). When sample, standard, and control containing HBsAg were added to the anti-HBs antibody-coated wells together with the anti-HBs peroxidase (horseradish) antibody and incubated, it led to the formation of antibody-HBsAg-antibody-peroxidase complex on the wells. After washing the microtiter plate to remove unbound material, a solution of TMB substrate was added to the wells and incubated. Color develops in proportion to the amount of HBsAg bound to anti-HBs. The peroxidase-TMB reaction is stopped by the addition of sulfuric acid. The optical density of developed color is read with a suitable photometer at 450 nm.

Anti-hepatitis C virus ELISA assay

This was carried out using AccuDiag™ HCV antibody ELISA kit. The manufacturer's instructions were strictly adhere to.

Sample, standard, and control were added to the polystyrene solid-phase wells precoated with HCV antigens. Moreover, they were incubated and washed followed by the addition of rabbit anti-human immunoglobulin M (IgM) antibodies (anti-IgM) conjugated to HRP-conjugate. The mixture was incubated again for the labeled antibodies to bind with any antigen-IgM complexes previously formed. The mixture was washed to remove unbound HRP conjugate followed by the addition of chromogen solutions containing TMB and urea peroxide to form a blue-colored solution in the presence of the antigen-antibody-anti-IgM (HRP) immunocomplex and when the colorless chromogens are hydrolyzed by the bound HRP conjugate. The reaction was stopped by the addition of sulfuric acid, which changed the blue color to yellow.

Method of statistical analysis

The results obtained were subjected to statistical analysis to determine mean, standard deviation, frequency, and percentage using the SPSS version 18.0 (IBM, Armonk, New York, USA). The frequency of occurrence and percentage values were used in the comparative analysis.

Ethical consideration

The proposal of this work was reviewed and approved by the Research and Ethical Committee of Baptist Medical Centre, Saki-Nigeria, before the commencement of the work. Consent of parents of each of the subjects was also obtained.


  Results Top


The viral immunochemical status obtained in children with elevated blood fibrinogen showed 3.7% (1) HIV mono-infection; 7.4% (2) HCV mono-infection; 18.5% (5) HBV mono-infection; 3.7% (1) HIV-HBV coinfection; and 7.4% (2) HCV-HBV coinfection with no coinfection of HIV-HCV and HIV-HCV-HBV, whereas the viral immunochemical status in hospital children showed only 11.1% (3) HBV mono-infection [Table 1] and [Figure 1].
Table 1: Viral immunochemical status of children aged 3-7 years with elevated and normal blood fibrinogen

Click here to view
Figure 1: (a) Stacked line chart of viral immunochemical status of children aged 3–7 years with elevated fibrinogen in herbal homes, hospitals, and children with normal blood fibrinogen. (b) Column chart of viral immunochemical status of children aged 3–7 years with elevated fibrinogen in herbal homes, hospitals, and children with normal blood fibrinogen

Click here to view


The viral immunochemical status obtained in children with normal blood fibrinogen showed 3.3% (1) HBV mono-infection and 3.3% (1) HCV mono-infection with no HIV mono-infection and HIV-HBV, HIV-HCV, HCV-HBV, and HIV-HCV-HBV coinfections [Table 1] and [Figure 1].


  Discussion Top


The viral immunochemical status obtained in children with elevated blood fibrinogen who received treatments in herbal homes showed 3.7% (1) HIV mono-infection; 7.4% (2) HCV mono-infection; 18.5% (5) HBV mono-infection; 3.7% (1) HIV-HBV coinfection; 7.4% (2) HCV-HBV coinfection with no coinfection of HIV-HCV and HIV-HCV-HBV compared with only 11.1% (3) HBV mono-infection in hospital children and in children with normal blood fibrinogen which showed 3.3% (1) HBV mono-infection and 3.3% (1) HCV mono-infection with no HIV mono-infection and HIV-HBV, HIV-HCV, HCV-HBV, and HIV-HCV-HBV coinfections.

Elevated blood fibrinogen in children could be associated with the fact that the coagulation and fibrinolysis physiology of newborn and young children significantly differ from older children and adults because of the immaturity of the hemostasis system of newborns. Healthy newborns also exhibit fewer symptoms of bleeding disorders and thrombosis.[18],[19]

The findings of this work as stated above could be attributed to the following that pathogenic microbes are triggers of inflammation. This immune response can cause increase or decrease in acute-phase proteins. Blood fibrinogen is a positive acute-phase protein whose blood levels increase in systemic inflammation as a result of pathogenic infections such as viral agents.[4],[5],[6],[7]

The HIV, HCV, HBV, and their coinfections found in children with elevated blood fibrinogen can be associated with immune defense activities of fibrinogen, which include trapping of the invading viral agents in the blood clots which might have induced liver for massive production and release of fibrinogen as a positive acute-phase protein as fibrinogen is made and secreted into the blood primarily by the liver hepatocytes.[1],[2],[3],[4],[5],[6],[7]

Viral seromarkers of HCV, HBV, HIV, HIV-HBV, and HCV-HBV were more in children with elevated blood fibrinogen than those with normal blood fibrinogen, which suggests blood fibrinogen as a possible diagnostic indicator in viral infection.

This difference could be linked with the explanation that pathogenic microbes can elicit inflammatory response indicated by elevated blood fibrinogen which may also reveal the degree of viral infection as shown in the pattern of viral seromarkers in tests and controls.[10],[11],[13]

The viral immunochemical status obtained in children with elevated blood fibrinogen who received treatments in herbal homes showed 3.7% (1) HIV mono-infection; 7.4% (2) HCV mono-infection; 18.5% (5) HBV mono-infection; 3.7% (1) HIV-HBV coinfection; and 7.4% (2) HCV-HBV coinfection with no coinfection of HIV-HCV and HIV-HCV-HBV compared with only 11.1% (3) HBV mono-infection obtained in the hospital children.

This could be attributed to the fact that hospital is a healthcare establishment providing patient treatment with specialized medical staff, equipment, and special consideration for control of infectious agents aimed at protecting healthcare givers and patients compared to herbal homes practice.[21]


  Conclusion Top


Viral seromarkers of HCV, HBV, HIV, HIV-HBV, and HCV-HBV were more in children with elevated blood fibrinogen who received treatments in herbal homes than those who received treatments in the hospital and those with normal blood fibrinogen, which suggests blood fibrinogen as a possible diagnostic indicator in viral infection in herbal homes.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

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Casjens S. Papillomaviruses of Animals. In: Mahy BW, Van Regenmortel MH. Desk Encyclopedia of General Virology. Boston: Academic Press; 2010. p. 167.  Back to cited text no. 10
    
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UNAIDS, WHO. 2007AIDS Epidemic Update (PDF). UNAIDS, WHO; 2007. p. 10. https://www.google.com/search?client=firefox-b-d&q=data.unaids.org+%E2%80%BA+pub+%E2%80%BA+epislides+%E2%80%BA+2007+%E2%80%BA+2007_epiupdate_en. [Last retrieved on 2008 Mar 12].  Back to cited text no. 14
    
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Revel-Vilk S. The conundrum of neonatal coagulopathy. Hematology Am Soc Hematol Educ Program 2012;2012:450-4.  Back to cited text no. 18
    
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Monagle P, Barnes C, Ignjatovic V, Furmedge J, Newall F, Chan A, et al. Developmental haemostasis. Impact for clinical haemostasis laboratories. Thromb Haemost 2006;95:362-72.  Back to cited text no. 19
    
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Cheesbrough M. District Laboratory Practice in Tropical Countries Part 2, 2nd ed. United States, America, New York: Cambridge University Press; 2006. Available from: http//www.cambridge.org/9780521676311.[Last accessed on 2019 Apr 12].  Back to cited text no. 20
    
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