Prevalence of HCV-Antibodies in Various Groups Screened for HBV at Khartoum Hospital

85 Pages·2003·1.11 MB·English

Prevalence of HCV-Antibodies

in Various Groups Screened for
HBV at Khartoum Hospital

By
Magda Mustafa Helmi
B. V. Sc. Faculty of Veterinary Sciences
University of Khartoum
June 2000

Supervisor:-Awad Al Karim A. Ibrahim
B. V. Sc. University of Khartoum
M. Sc. A'dam University, The Netherlands
Ph. D. Utrech State University, The Netherlands

Co-supervisor:-Shamsoun K. Kafi
M.B.B.S. University of Khartoum
M. D. University of Khartoum

A Thesis Submitted to the University of Khartoum
in Partial Fulfillment of the Requirements for Master
Degree of Microbiology

June 2003
ﻢﻴﺣﺮﻟا ﻦﻤﺣﺮﻟا ﷲا ﻢﺴﺑ

ﻰﻟ ﺮﺴﻱ و , ىرﺪﺻ ﻰﻟ حﺮﺷا بر لﺎ(cid:1127)ﻗ ) : ﻰﻟﺎ(cid:1127)ﻌﺗ لﺎ(cid:1127)ﻗ

( ﻰﻟﻮﻗ اﻮﻬﻘﻔﻱ , ﻰﻧﺎﺴﻟ ﻦﻡ ةﺪﻘﻋ ﻞﻠﺣا و , ىﺮﻡا

ﻢﻴﻈﻌﻟا ﷲا قﺪﺻ
ﻪﻃ ةرﻮﺱ 28-25 تﺎﻱﻻا

2
DEDICATION

To My Father, Mother, Sister,

And Who Care for Me.

3
ACKNOWLEDGMENT

Praise to Allah Who gave me health and power
to do this work.
I am heavily indebted to my supervisor Dr.
Awad El Karim A. Ibrahim, head of Microbiology
department, University of Khartoum; and my co-
supervisor Dr. Shamsoon K Kafi, head of Virology
department, National Health Laboratory, for their
help, encouragement, support and patience,
without whom this research would have never been
done.
I would like to thanks all those at the Virology
department, at the National Health Laboratory,
for their generous hospitality and co-operation
which was very important to complete this work.
Thanks are also due to those at Khartoum
Center for Kidney Transplant and Dialysis; and
those at the blood bank of Khartoum's Hospital
for their help.
Special thanks to Professor Hassan Sid
Ahmed for his assistance at the beginning phase
of this research.
At last and not least, great thanks to my
relatives, friends and my colleagues at the
Microbiology department, University of Khartoum,
for their encouragement.

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CONTENTS
Dedication………………………………………………………………..3
Acknowledgements ……………………………………………………...4
Contents ………………………………………………………………….5
List of Tables …………………………………………………………….7
List of Figures …………………………………………………………...8
Abstracts ………………………………………………………………....9
Arabic abstracts ………………………………………………………...10
Chapter One: Introduction ……………………………………………..11
Chapter Two: Literature Review ……………………………..13
2.1 Definition ………………………………………………………….13
2.2 Aetiology …………………………………………………………..13
2.3 Viral hepatitis ………………………………………………………14
2.3.1 Hepatitis type A ………………………………………………….14
2.3.2 Hepatitis type B ………………………………………………….16
2.3.3 Hepatitis type D ………………………………………………….16
2.3.4 Hepatitis type E …………………………………………………..17
2.3.5 Hepatitis type G…………………………………………………..17
2.4 Hepatitis type C ……………………………………………………17
2.5 HCV classification …………………………………………………19
2.6 Genetic structure of HCV ………………………………………….21
2.7 HCV replication cycle ……………………………………………..23
2.8 Epidemiology of HCV ……………………………………………..24
2.8.1 Modes of transmission……………………………………………24
2.8.1.1 Blood …………………………………………………………..24
2.8.1.2 Vertical transmission …………………………………………..25
2.8.1.3 Sexual transmission…………………………………………….25
2.8.1.4 Occupational exposure …………………………………………25
2.8.1.5 Haemodialysis ………………………………………………….26
2.8.1.6 Body fluids……………………………………………………...26
2.8.1.7 Other modes of transmission …………………………………..27
2.8.2 Seroprevalence …………………………………………………...27
2.8.3 Genotypic prevalence ……………………………………………31
2.8.4 HCV prevalence in Sudan ……………………………………….33
2.9 Clinical presentation ……………………………………………….34
2.9.1 Laboratory feature and biochemical abnormalities ……………...36
2.9.2 Pathological features ……………………………………………..38
2.9.2.1 Classical acute hepatitis ………………………………………..38
2.9.2.2 Acute hepatitis with bridging necrosis ………………………...40
2.9.2.2.1 Acute hepatitis with massive necrosis ………………………40
2.9.2.3 Progression to chronic liver disease ……………………………40
2.9.2.4 Liver Failure ……………………………………………………41
2.9.2.5 Histologic abnormalities ………………………………………45
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2.9.2.6 Hepatocellular carcinoma ………………………………………47
2.10 Diagnosis of HCV infection ………………………………………48
2.10.1 Immunodiagnostic tests ………………………………………...48
2.10.1.1 The anti-hepatitis C test ………………………………………48
2.10.1.2 Recombinant Immuno Blot Assay ……………………………51
2.10.2 Nucleic acid probe test ………………………………………….52
2.10.2.1 Polymerase Chain Reaction …………………………………..52
2.10.2.2 Genotype test …………………………………………………53
2.10.2.3 Viral Load Test ………………………………………………53
2.10.3 Other laboratory and clinical tests ……………………………..54
2.10.3.1 Ultrasound scan of liver ………………………………………54
2.10.3.2 Liver function tests …………………………………………...54
2.10.3.2.1 Transaminase values ………………………………………..54
2.10.3.2.2 Bilirubin ……………………………………………………55
2.10.3.2.3 Albumin …………………………………………………….55
2.10.3.2.4 Prothrombin Time (PT) …………………………………….55
2.10.3.3 Full blood count ………………………………………………55
2.11 Treatment …………………………………………………………55
2.11.1 Side effects ……………………………………………………..56
2.12 Prevention ………………………………………………………..57
Chapter Three: Material and Methods …………………………………58
3.1 Collection of samples ……………………………………………...58
3.2 Indirect ELISA for the detection of HCV-Abs ……………………59
3.2.1 ELISA kits ……………………………………………………….59
3.2.2 Other materials not included in the kits………………………….60
3.3 Storage of the ELISA kits ………………………………………….60
3.4 Assay procedure ……………………………………………………60
3.5 Test procedure………………………………………………………61
3.6 Reading of the test …………………………………………………62
3.7 Interpretation of the test ……………………………………………63
Chapter Four: Results ………………………………………………….64
4.1. Blood donors group ……………………………………………….64
4.2. Travelers group ……………………………………………………64
4.3. Haemodialysis group ……………………………………………...64
4.4. Jaundiced patients group ………………………………………….65
4.5 Optical Density obtained……………………………………………65
4.6 Risk factors that contribute to HCV transmission …………………65
chapter Five: Discussion ………………………………………………70
5.1 Conclusion ………………………………………………………...73
5.2 Recommendations …………………………………………………74
References ……………………………………………………………..75
Appendix ………………………………………………………………83

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List of Tables
Table (1): Characteristics of hepatitis viruses (Jawetz et al 1998) …….15
Table (2): Nomenclature and definitions of hepatitis viruses, antigens and
antibodies (Jawetz et al1998)...............................................18
Table (3): Classification of HCV isolates based on phelogenetic analysis
(Forms &Bukh 1999)...............................................................20
Table (4): Epidemiologic and clinical features of viral hepatitis types A, B
and C (Jawetz et al1998)......................................................35
Table (5): Aetiological types of chronic active hepatitis (Mowat et al
2001)………………………………………………………… 42
Table (6): Total HCV positive & HCV negative in tested roups………66
Table (7): Optical Density of each HCV positive sample……………..66
Table (8): Data obtained from the uestionnaire………………………..66
Table (9): Risk factors found with HCV positive amples……………...68
Table (10): Relative Risk for each risk factor…………………………....69

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List of Figures
Fig. (1): Genetic organization of the HCV genome (Jawetz et al 1998).22
Fig. (2): Hepatitis C virus (HCV) prevalence in the world (Cohen1999)28
Fig. (3): Worldwide distribution of Various Genotypes of Hepatitis C
Virous (forms & Bukh 999)……………………………………28
Fig. (4a): Liver is divided histologically into lobules (Website )………39
Fig. (4b): Collapse of the liver parenchyma with viral hepatitis C
(Website 4 )…………………………………………………39
Fig. (5a): Viral hepatitis C with extensive fibrosis ( Website 4 )……....43
Fig. (5b): The regenerative nodules of hepatocytes are surrounded by
Fibrous connective tissue (Website 4 )………………………43
Fig. (6a): Viral hepatitis C which leads to chronic liver disease (Website
4 )……………………………………………………………46
Fig. (6b): Malignant cells of hepatocellular carcinoma.(Website 4)….46
Fig. (7): Algorithm for the serologic diagnosis of HAV, HBV, HCV and
HDV.(Jawetz et al 1998 )………………………………………49
Fig. (8):Optical Density variation between HCV positive samples……66

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ABSTRACT
This study was done to evaluate the seroprevalence of HCV-Abs in
Khartoum area. Four hundred samples were screened for the presence of
HCV-Abs by indirect ELISA using recombinant HCV-Ags. Samples were
collected randomly from different groups including: 150 blood samples
from blood donors at the blood bank of Khartoum Hospital; 100 blood
samples were collected from travelers coming to the National Health
Laboratory being tested for other viral infections; 40 blood samples were
collected from haemodiolysis patients at Khartoum Center for Kidney
Transplant and Dialysis and 110 blood sample from jaundiced patients
visiting the Central National Health Laboratory. From this group 60
samples were previously diagnosed as HBV positive and 50 samples were
HBV negative.
All blood donors and travelers were found negative for HCV-Abs In
contrast 15 patients out of the 40 of the haemodialysis group were found
positive. This represents 37.5% of this group and 3.75% of the total number
of samples being investigated as positive for the presence of HCV-Ab. In
jaundiced patients only 3 patients were found positive for HCV-Abs. This
gave 2.7% of this group and 0.75% of the total samples tested. When
compared with the risk factors it was found that 83.33% of the HCV-Abs
positive patients have been exposed to haemoclialysis machines while the
other 16.66% of HCV-Abs patients were exposed to other risk factors.
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ﺔﺣﻭﺮﻃﻷﺍ ﺺﺨﻠﻣ
ﺏﺎﻬﺘﻟﻹﺍ ﺽﺭﻤﻟ ﺓﺩﺎﻀﻤﻟﺍ ﻡﺎﺴﺠﻷﺍ ﺭﺎﺸﺘﻨﺇ ﻱﺩﻤ ﺔﻓﺭﻌﻤﻟ ﻡﻭﻁﺭﺨﻟﺍ ﺔﻘﻁﻨﻤ ﻲﻓ ﺔﺴﺍﺭﺩﻟﺍ ﻩﺫﻫ ﺕﻴﺭﺠﺃ
ﺽﺭﻤﻟ ﺓﺩﺎﻀﻤﻟﺍ ﻡﺎﺴﺠﻷﺍ ﺩﻭﺠﻭ ﺔﻓﺭﻌﻤﻟ ﻡﺩ ﺔﻨﻴﻋ 400 ﻲﻠﻋ ﺭﺎﺒﺘﺨﻹﺍ ﻱﺭﺠﺃ .(ﺝ) ﻉﻭﻨ ﻥﻤ ﻱﺩﺒﻜﻟﺍ
ﺱﻭﺭﻴﻔﻟﺍ ﺕﺎﻨﻴﺠﺘﻨﺃ لﺎﻤﻌﺘﺴﺎﺒ ﺭﺸﺎﺒﻤﻟﺍ ﺭﻴﻏ ﺍﺯﻴﻟﻹﺍ ﺭﺎﺒﺘﺨﺍ ﺔﻁﺴﺍﻭﺒ (ﺝ) ﺱﻭﺭﻴﻓ ﻱﺩﺒﻜﻟﺍ ﺏﺎﻬﺘﻟﻹﺍ
ﺕﺎﻋﻭﻤﺠﻤ ﻥﻤ ﺎﹰﻴﺌﺍﻭﺸﻋ ﺕﺎﻨﻴﻌﻟﺍ ﻊﻤﺠ ﻡﺘ .(Recombinant-Ag ﻥﻴﺠﻫ) ﺎﹰﻴﺜﺍﺭﻭ ﺓﺭﻀﺤﻤﻟﺍ (ﺝ)
ﺔﻨﻴﻋ 100 ؛ﻡﻭﻁﺭﺨﻟﺍ ﻲﻔﺸﺘﺴﻤ ﻲﻓ ﻡﺩﻟﺍ ﻙﻨﺒ ﻲﻓ ﻡﺩﻟﺎﺒ ﻥﻴﻋﺭﺒﺘﻤﻟﺍ ﻥﻤ ﻡﺩ ﺔﻨﻴﻋ 150 لﻤﺸﺘ ﺔﻔﻠﺘﺨﻤ
ﺽﺍﺭﻤﻷ ﻡﻬﺼﺤﻓ ﻡﺘ ﻥﻴﺫﻟﺍ ﻱﺯﻜﺭﻤﻟﺍ ﻲﻤﻭﻘﻟﺍ ﻲﺤﺼﻟﺍ لﻤﻌﻤﻠﻟ ﻥﻴﺭﺌﺍﺯﻟﺍ ﻥﻴﺭﻓﺎﺴﻤﻟﺍ ﻥﻤ ﺕﻌﻤﺠ ﻡﺩ
ﺔﻋﺍﺭﺯﻟ ﻡﻭﻁﺭﺨﻟﺍ ﺯﻜﺭﻤ ﻲﻓ ﻱﻭﻠﻜﻟﺍ لﺸﻔﻟﺍ ﻲﻀﺭﻤ ﻥﻤ ﺕﻌﻤﺠ ﻡﺩ ﺔﻨﻴﻋ 40 ؛ﻱﺭﺨﺃ ﺔﻴﺴﻭﺭﻴﻓ
ﻥﻴﺭﺌﺍﺯ ﻲﻀﺭﻤ ﺽﺭﻌﻜ ﻥﺎﻗﺭﻴﻟﺍ ﻥﻤ ﻥﻭﻨﺎﻌﻴ ﻲﻀﺭﻤ ﻥﻤ ﺕﻌﻤﺠ ﻡﺩ ﺔﻨﻴﻋ 110 ﻭ ﻲﻠﻜﻟﺍ لﻴﺴﻏﻭ
،(ﺏ) ﺱﻭﺭﻴﻓ ﻱﺩﺒﻜﻟﺍ ﺏﺎﻬﺘﻟﻹﺍ ﺽﺭﻤ ﺩﻀ ﺎﹰﻘﺒﺴﻤ ﺎﻬﺼﺤﻓ ﻡﺘ ﻱﺯﻜﺭﻤﻟﺍ ﻲﻤﻭﻘﻟﺍ ﻲﺤﺼﻟﺍ لﻤﻌﻤﻠﻟ
ﺕﺩﺠﻭ ﺔﻨﻴﻋ 50 ﺎﻤﻨﻴﺒ (ﺏ) ﺱﻭﺭﻴﻓ ﻱﺩﺒﻜﻟﺍ ﺏﺎﺘﻟ ﻹﺍ ﺽﺭﻤﻟ ﺔﺒﺠﻭﻤ ﺎﻬﻌﻴﻤﺠ ﺕﺩﺠﻭ ﻡﺩ ﺔﻨﻴﻋ 60
.ﻪﻟ ﺔﺒﻟﺎﺴ ﺎﻬﻌﻴﻤﺠ
ﻲﻓ .(ﺝ) ﺱﻭﺭﻴﻓ ﻱﺩﺒﻜﻟﺍ ﺏﺎﻬﺘﻟﻹﺍ ﺽﺭﻤﻟ ﺔﺒﻟﺎﺴ ﺕﺩﺠﻭ ﻥﻴﺭﻓﺎﺴﻤﻟﺍﻭ ﻡﺩﻟﺍ ﻲﻋﺭﺒﺘﻤ ﺕﺎﻨﻴﻋ ﻊﻴﻤﺠ
ﺱﻭﺭﻴﻓ ﻱﺩﺒﻜﻟﺍ ﺏﺎﻬﺘﻟﻹﺍ ﺽﺭﻤﻟ ﺔﺒﺠﻭﻤ ﺕﺩﺠﻭ ﻱﻭﻠﻜﻟﺍ لﺸﻔﻟﺍ ﻲﻀﺭﻤ ﻥﻴﺒ ﻥﻤ ﺔﻨﻴﻋ 15 لﺒﺎﻘﻤﻟﺍ
ﺕﺎﻨﻴﻌﻟﺍ ﻉﻭﻤﺠﻤ ﻥﻤ %3.75 لﺜﻤﺘ ﻥﻴﺤ ﻲﻓ ﺔﻋﻭﻤﺠ ﻤﻟﺍ ﻩﺫﻫ ﻥﻤ %37.5 لﺜﻤﺘ ﺔﺠﻴﺘﻨﻟﺍ ﻩﺫﻫ .(ﺝ)
ﺩﻘﻓ ﻲﻀﺭﻤ ﺽﺭﻌﻜ ﻥﺎﻗﺭﻴﻟﺍ ﻥﻤ ﻥﻭﻨﺎﻌﻴ ﻥﻴﺫﻟﺍ ﻲﻀﺭﻤﻠﻟ ﺔﺒﺴﻨﻟﺎﺒ . ﺔﺴﺍﺭﺩﻟﺍ ﻩﺫﻫ ﻲﻓ ﺎﻬﺼﺤﻓ ﻡﺘ ﻲﺘﻟﺍ
ﻥﻤ %2.7 ﺔﺒﺴﻨ لﺜﻤﺘ ﻩﺫﻫ ،(ﺝ) ﺱﻭﺭﻴﻓ ﻱﺩﺒﻜﻟﺍ ﺏﺎﻬﺘﻟﻹﺍ ﺽﺭﻤﻟ ﺔﺒﺠﻭﻤ ﻁﻘﻓ ﺕﺎﻨﻴﻋ 3 ﺕﺩﺠﻭ
لﻤﺍﻭﻌﻟﺎﺒ ﺔﻨﺭﺎﻘﻤﻟﺍ ﺩﻨﻋ . ﺎﻬﺼﺤﻓ ﻡﺘ ﻲﺘﻟﺍ ﺕﺎﻨﻴﻌﻟﺍ ﻉﻭﻤﺠﻤ ﻥﻤ %0.75 ﺔﺒﺴﻨﻭ ﺔﻋﻭﻤﺠﻤﻟﺍ ﻩﺫﻫ
ﺽﺭﻤﻟ ﻲﺒﺠﻭﻤ ﻲﻀﺭﻤﻟﺍ ﻉﻭﻤﺠﻤ ﻥﻤ %83.33 ﻥﺃ ﺩﺠﻭ ﺩﻘﻓ ﺽﺭﻤﻟﺍ ﺭﺎﺸﺘﻨﺇ ﻲﻠﻋ ﺓﺩﻋﺎﺴﻤﻟﺍ
ﻉﻭﻤﺠﻤ ﻥﻤ %16.66 ﺎﻤﻨﻴﺒ ﻱﻭﻠﻜﻟﺍ لﻴﺴﻐﻟﺍ ﺕﺎﻨﻴﻜﺎﻤﻟ ﺍﻭﻀﺭﻌﺘ ﺩﻗ (ﺝ) ﺱﻭﺭﻴﻓ ﺩﺒﻜﻟﺍ ﺏﺎﻬﺘﻟﻹﺍ
.ﺔﻔﻠﺘﺤﻤ ﻱﺭﺨﺃ لﻤﺍﻭﻌﻟ ﺍﻭﻀﺭﻌﺘ ﺩﻗ ﻲﻀﺭﻤﻟﺍ
10