Incidence Of Hepatitis B Virus And Risk Factors Biology Essay

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Hepatitis B virus is highly endemic in Pakistan. It may cause a wide variety of acute or chronic hepatic and extrahepatic diseases, as well as chronic carrier state.The purpose of this study was to identify HBV and its risks factors in the general population of Thall.


This study was conducted at Al-Rehman Medical Laboratory from February 2011 to April 2011. A total of 200 blood samples were tested for HBV. All blood samples were initially tested by using Immunochromatograhic Technique (ICT) and confirmed by using 3rd Generation Enzyme Immunosorbent Assay (ELISA). The results of ICT and ELISA were also compared.


Among 200 blood samples, 07 cases were positive for Hepatitis B surface antigen (HBsAg). These 07 cases had different histories of dental treatment (42.85%), traveled abroad (28.57%), surgery (14.28%) and blood transfusion. Out of these 07 cases, one sample was negative on Immunochromatographic Technique (ICT), while all 07 the samples were positive on ELISA 3rd Generation Method. These results suggested that the rapid immuno-chromatographic kit for HBsAg has only limited efficacy and must be confirmed through superior method like ELISA.


The number of positive cases of Hepatitis B virus were 07 (3.5%) in 200 blood samples collected from different areas of Thall. It can be concluded that 3rd Generation ELISA is more specific and sensitive than ICT and shows good results.Dental treatment (42.85%) was the main risk factor for transmission of HBV infection. The preventive strategies should be adopted for Hepatitis B virus which constitutes a major public health problem in Pakistan.

Abbreviations and Acronyms

HBV Hepatitis B Virus

HCV Hepatitis C Virus

RNA Ribo Nucleic Acid

ELISA Enzyme Linked Immunosorbent Assay

MEIA Microparticle Enzyme Immunoassay

ALT Alanine Aminotransfrase

PCR Polymerase Chain Reaction

WHO World Health Organization

CRF Chronic Renal Failure

TTVI Transfusion-Transmissible Viral Infection

DNA Deoxyribo Nucleic Acid

OR Odds Ratios

HBsAg Hepatitis B surface antigen

HBeAg Hepatitis B e antigen

Anti-HBe Anti- Hepatitis B e antigen

HBcAg Hepatitis B core antigen

EIA Enzyme immunoassay

RIBA Recombinant Immunoblot Assay

ICT Immunochromatographic Technique

ml Mili liters

µl Micro liters

TMB T Etramethyl-Benzidine

NAT Nucleic acid testing

N Number

IgG Immunoglobulin G

°C Centigrade


Certificate of Approval I

Declaration ii

Acknowledgement iv

Introduction v

Methods v

Results v

Conclusion vi

Abbreviations and Acronyms vii

Contents viii

Chapter 1 1


Chapter 2 4


Chapter 3 14




Chapter 4 18


Chapter 5 22


Chapter 6 25







Questionnaire 32


Consent form for the participants 33

Chapter 1


Hepatitis B is found worldwide but is particularly prevalent in Asia, globally about 350 million people are chronically infected with HBV and about 75% of them are Asian. Hepatitis B virus causes hepatitis infection which is complex structure double stranded DNA virus. The envelope contains a protein called the surface antigen (HBsAg). There are two other important antigens, which are the core antigen and the e antigen .The core antigen is known as (HBcAg) and the e antigen is known as (HBeAg). HbeAg is an important indicator of transmissibility because the e antigen is located on the core protein, which means the infectious particle must be present in the patient's blood (Warren Levinson, 2004).

The prevalence of hepatitis B virus have different range in different areas of the world (Lavanchy, 2004), In the middle east, sub-Saharan Africa and far east the prevalence show the high rate ranging to greater than 8% positivity while in the developed countries like Northern Europe, United States and in Australia the ratio is intermediate which is ranging from 2% to 7%. In some other countries like India and Japan and the areas of central Asia(Andre,2000).Study shows that prevalence rate in Pakistan is 35% to 38% in which 4% are count as the carrier (Qureshi, 2003). In Karachi during 2002 to 2006, 351309 samples of males blood were screened in which the prevalence rate of hepatitis B rate was 2% (Akhtar, 2005), the health care personals have 2.4% prevalence rate in Karachi health (Khichi and Channar, 2000), s In Abbottabad the prevalence rate was 2.68% in 5,207 donors (Ally et al., 2005). In Pakistan 903 afghan refugees were screened having 8.3% prevalence rate (Quddus et al., 2006)). According to Zuberi (1996) the 10% of healthy adults were found the HBV carriers.

There are three main modes of transmission, via sexual intercourse, parentally from mother to new born and blood transfusion. It is concluded that the 5% of patients with HBV infection become chronic carrier i.e. someone who has HBsAg persisting in their blood for at least six months (Warren Levinson, 2004). Vertical transmission from infected mother to their new born child is of great importance. Women's those having HBs Ag carriers can transmit the virus; however, but the degree of the infection is different from individual to individual. The transmission of HbeAg is greater in infected mothers as compared with HBe. HBV infection in infancy plays a key role which leads to chronicity. The infants with 2 to 3 months, goes to the carrier stage with the percentage ranging from 80% to 85% (Kazmi et al., 2003).

Risk factors by which the Hep B spread are the direct contact with Hep B infected person, unsafe sexual contacts, through intravenous drug uses, through different surgical and dental procedures (Sali et al., 2005). The chronic carrier of hepatitis B leads to serious diseases, like chronic hepatitis, liver cirrhosis and hepatocelluler carcinoma (Dragosics et al., 1987).

It has been observed in Nepal that Hepatitis B is an important public health issue in blood donors with acute hepatitis rate of 6%. About 1% of the populations are asymptomatic chronic HBsAg carrier, while 39% of patients suffering from chronic liver disease and 37% with hepatocelluler carcinoma are HBsAg seropositive (Karki et al., 2008).

In the same scenario, this study has been designed to identify HBV, acute infection (active) and chronic infection (carrier state) in blood donors in our local environment.


To identify Hepatitis B in blood donors.

To find out the active infection and carrier state in blood donors who are the source of infection for others.

To compare the results of ICT and ELISA Method for identification of HBV.

Chapter 2


Khan et al. 2010 conducted a study at the King Edward Medical University, Lahore during the period between November 2009 and January 2010. Four rapid immuno-chromatographic assays - ONE STEP HBsAg One Check, One step HBsAg test Accurate were evaluated for detection of hepatitis B surface antigen. Total of 57 tested for Hepatitis B by ELISA were stored at -20 °C and subsequently tested with rapid ICT tests (devices) (Accurate and One Check brands). Sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy of these tests were calculated using ELISA as the gold standard. This study was conducted to evaluate the performance of two commercially available rapid immuno-chromatographic testing (ICT) kits for HBsAg. Out of 38 positive samples by ELISA, 20 samples were positive for HBsAg by one check and 19 samples were positive by accurate. The sensitivity for one check was found to be 53% and 50 % for accurate. The negative predictive value for one check was 51% and 49% for Accurate. The positive predictive values for HBV were 100% for one check and 95% with Accurate.

Ali et al. 2009 evaluated that Pakistan carries one of the world's highest burdens of chronic hepatitis and mortality due to liver failure and hepatocellular carcinomas. However, national level estimates prevalence and risk factors for hepatitis B. They observed the medical and public health literature over a 13-year period (1994-2007) to evaluate the prevalence of active hepatitis B and chronic hepatitis C in Pakistan. They analyzed data separately for the general and high-risk populations for each of the four provinces. A weighted average of hepatitis B antigen prevalence among healthy adults (blood donors and non-donors) was 2.4% (range 1.4-11.0%). Data suggest a moderate to high prevalence of hepatitis B and hepatitis C in different areas of Pakistan. The published literature on the modes of transmission of hepatitis B in Pakistan implicate contaminated needle use in medical care and drug abuse and unsafe blood and blood product transfusion as the major causal factors.

Ogbu and Uneke, 2009 observed that Hepatitis B virus (HBV) is the most common cause of serious liver infection in the world. It is estimated that worldwide more than two billion people infected by HBV and 350 million people have chronic infection. The HBV is highly contagious and transmission occurs through percutaneous or permucosal routes, and infective blood or body fluids, through sexual contact or by contaminated needles. In sub-Saharan Africa, transfusion-transmitted HBV infection is increasingly becoming a major mode of transmission of HBV in the high-prevalence areas. The demand for blood transfusion services is high and increase the possibility of the transmission of HBV through contaminated blood Due to endemicity of infections causing anemia, malnutrition, and surgical and obstetrical emergencies associated with blood loss in the sub-Saharan Africa. Blood safety remains a big issue in transfusion medicine in this part of the world because national blood transfusion services and policies, appropriate infrastructure, trained personnel and financial resources are inadequate. As part of public health interventional measures, the transmission of HBV can be minimized by pre-screening donation, by evaluation of high-risk donors.

Alavian et al. 2008 observed that Hepatitis B virus (HBV) infection is a global problem. It is estimated that 400 million people are suffering from this infection. They reviewed to put all evidence on HBV infection in Iran and to make an accurate estimate of HBV infection prevalence in Iran for further planning to control the infection. Meta-analysis and survey data analysis of all national and international papers, theses, congresses, reports, Iranian medical universities projects, research centers, reports of Deputy for Health affairs (published or unpublished) Iranian general population with positive HBsAg in blood samples. All descriptive/analytical cross-sectional studies/surveys from April 2001 to March 2007 that have sufficiently declared objectives, proper sampling method with identical and valid measurement instruments for all study subjects and proper analysis methods regarding sampling design and demographic adjustments Presence of positive HBsAg in blood samples of study samples Fourteen studies met the inclusion criteria. They were from 7 (out of 30) provinces in which about 40 percent of the country population live. These provinces (HBsAg positive prevalence) were Golestan (6.3%), Tehran (2.2%), East Azarbaijan (1.3%), Hamedan (2.3%), Isfahan (1.3%), Kermanshah (1.3%) and Hormozgan (2.4%). The HBV infection prevalence in Iran is estimated to be 2.14 percent, in men and women 2.55 percent and 2.03 percent respectively. About 1.5 million people in Iran are living with HBV infection and it is estimated that 15% to 40% of them are at risk of developing cirrhosis and/or hepatocellular carcinoma (HCC) without intervention. The prevalence of HBV infection has been reported higher in more recent studies compared to the study in 2000-2001.

Behal et al. 2008 determined that Infection with hepatitis-B virus has been a significant cause of morbidity claiming more than a million lives every year. Epidemiological data reveals that there are 360 million carriers of hepatitis-B virus throughout the globe and 78% of the world populations' hail from Asia. Though several studies from Indian sub-continent have provided an estimate of the prevalence of this viral infection, there exist only few studies, which reflect the status in the general population. The present study was conducted to investigate the prevalence of hepatitis-B infection in North Indian general population. A total of 20,000 healthy blood donors were screened for hepatitis-B surface antigen (HBsAg) status using third generation ELISA kit. Seroprevalence rate of seropositive donors was calculated and classified by age, sex and blood groups. Statistical analysis was done by using tests of proportions, chi-square and confidence interval. Out of 20,000 donors, 450 (2.25%) were HBsAg positive (95% confidence interval (CI), 2.0445-2.4554). Higher prevalence of HBsAg was found among males (440/19235) than females (10/765).

In 2008, Tanveer et al. conducted a study in the new campus of Punjab University Lahore. In the study they checked the students and administrative staff for Hepatitis B antigens (Hbs Ag) and antibodies for Hepatitis C virus (anti- HCV). The prevalence rate of anti-HCV was found 1.48 % and HBsAg was 2.46 % respectively. it is essential that all donors should be screened for anti- HCV and HbsAg in order to prevent the transmission of HBV and HCV through blood transfusion. It is necessary to awake the awareness about it.

It was observed in 2007 by Alavian et al. that during the last decade, the prevalence of Hepatitis B virus (HBV) has decreased in Iranian population. The reason for this decrease in the prevalence of Hepatitis B is probably the increase in awareness about HBV risk factors, national vaccination program since 1993 for all neonates, and vaccination of high risk groups. Hepatitis B vaccination was included in the EPI in 1993 and after 13 years of its implementation, the coverage has reached considerably from 62% in 1993 to 94% in 2005. With immunization of high risk people for Hepatitis B virus, surveillance of individuals infected with hepatitis B and control on the health status of refugees will further lower the rate of Hepatitis B infection in Iran. It is very important to control all the possible routes of transmission of Hepatitis B virus in order to control its spread. Practice of Universal precautions in the health institutions plays a key role in the spread of Hepatitis B and other dangerous viral infections.

In 2007, it was reported by Cohen, that certain viruses called hepatotropic virus inflame the live of the subject, which is diffuse in nature and is called acute viral hepatitis. Other than nonspecific symptoms of viral diseases, there can be nausea, vomiting and even loss of appetite. Sometimes it is associated with fever as well as pain in the abdomen particularly in the right upper quadrant. Later on when other signs and symptoms disappear, yellow discoloration of the skin and sclera of the eyes appear which is called jaundice. Majority of the case resolve but in rare cases, chronic hepatitis develop.

Hepatitis B is diagnosed by the identification of virus in the body, via different tests on blood. Far prevention of the viral hepatitis, personal hygiene is very important. Pre exposure and post exposure prophylaxis may be possible using vaccines or serum globulins depending on specific virus. There is no specific treatment, just supportive measures are taken and symptoms are treated accordingly.

Globally acute viral hepatitis is very common. There is variety of causes of this viral infection.

In 2007, Essa and Farhan, 2007 pointed out that viral hepatitis is main problem in Pakistan and HBV is highly antigenic. In this study a total of 35,257 subjects of including males as well females were screened, where HBsAg has to be detected. The study lasted for eight years from which ended in 2006.

In many Asian countries Hepatitis B virus is endemic as observed by Liuac et al. in 2006. Transfusion is the most common route among many transmission routes that should be prevented. In early 1970s, hepatitis B surface antigen was implemented, which was no doubt a great success in enhancing the safety of transfusion. However, it was demonstrated that transmission of the said virus by blood negative for HBsAg can still be possible in the early stages of infection or during chronic stages of infection (i.e. "occult" HBV infection, OHB) (Liuac et al. in 2006). OHB is defined as the presence of HBV DNA in blood or liver tissues in patients negative for HBsAg, with or without any HBV antibodies. Because of limitations in current blood screening practices, OHB is an overlooked source of HBV transmission. New policy for screening should be formulated and imposed on the basis of available data or newly designed studies.

In order to measure the level of HBsAg on ELISA technique, a study was carried out in Jordan. The said study was conducted in 1997 on 247 children who did not get their vaccination. I in 2004 247 more kids, who had their vaccination done, were studied. A total of 8 persons are HBs-Ag positive and only one child (0.4) is positive from the vaccinated group.

In 2003, Bhatta et al. performed a study in Khatmandu in a tertiary care hospital. The total duration of study was nine months from April to December. The objective was to measure the prevalence of Hepatitis B in the patients coming to the teaching hospital for treatment purposes. 200 patients participated in the study. Blood from the participants was collected and tested for the presence of Hepatitis B virus. Amongst all, total 5 were tested positive all of whom were male of young age.

In 2003, in a study the researcher Roger et al. obtained epidemiologic and serologic data on 422 household contacts of 157 blood donor candidates in the Baltimore Regional Red Cross Blood Program who were asymptomatic chronic carriers of hepatitis B surface antigen (HBsAg) (index carriers), and on 318 household contacts of 157 blood donors who were negative for serologic markers of hepatitis B virus (index controls). Index carriers were matched to index controls and their household contacts were similar with respect to race, sex, and education level completed. HBsAg prevalence was 6.8 times higher, and antibody to HBsAg (anti-HBs) prevalence 2.7 times higher among the contacts of carriers. The combined prevalence for hepatitis B virus markers was 29.9% among the contacts of carriers compared to 8.8% among the contacts of controls (p < 0.001). Being non-white, the sibling of a carrier, of older age, of lower educational status, and a member of a small household were significantly associated with either a higher antibody or combined hepatitis B prevalence rate (Roger et al. 2003).The contacts of female carriers had an HBsAg prevalence rate 4.7 times greater than the contacts of male carriers (Roger et al. 2003). Children who had a mother as their index carrier had an eleven fold higher HBsAg prevalence than did children who had a father as their index carrier (18.0% vs. 1.6%, p <0.005) (Roger et al. 2003). Epidemiologic characteristics of the contacts or their Index carriers, such as the sharing of Items belonging to the index carrier, the type and frequency of sexual relations, and the frequency of potential exposure to body fluids of the index carriers were examined. Contacts of carriers who had had skin lesions in the past year had a higher prevalence of serologic markers for hepatitis B than contacts of carriers without such lesions. (Roger et al. 2003). No sex-related factors among spouses were significantly associated with an increased prevalence of serologic markers (Roger et al. 2003). These data give suggest that person-to-person modes of transmission are important in the spread of hepatitis B within households. The patterns of hepatitis B serologic markers observed here suggest that mother to child transmission is most important in accounting for the high HBsAg prevalence among the contacts; other presently unidentified but close types of contact best explain the patterns of antibody prevalence.

Hussain et al conducted a study in 1998 in Pakistan. According to his study, Hepatitis B virus (HBV) is very much endemic in Pakistan, especially in rural areas. It can be the cause of all types of Hepatitis, like acute hepatitis, chronic hepatitis. It may also result in fulminate hepatic failure or may even take the form into carrier state.

Franchis et al. found in 1993 that those carrying surface antigen of hepatitis b with normal liver function tests, took into account the carcinoma of hepatic cells and other serious live diseases. Majority (69) of the participants had normal findings on the basis of histology while 18 individuals had chronic hepatitis of serious nature and 5 were having mild disease of the live. Most the participants had Serum enzyme levels within normal limits which was 58 of 68 participants who had regular follow-up. Out of the all 68 participants, 3 had chemical changes which were associated to hepatitis B virus (HBV) infection (Franchis et al.) In one of these patients, a later histologic evaluation showed progression to chronic active hepatitis (Franchis et al.). One patient had developed alcoholic cirrhosis (Franchis et al.). Ten patients showed loss of HBsAg; 2 of these patients acquired antibody to hepatitis B surface antigen (anti-HBs) (Franchis et al.). All patients who did not have regular follow-up, except 1, were interviewed by telephone during 1990 (Franchis et al.). All of them were accepting that they had liver disease of any kind. None of the subjects had carcinoma of hepatic cells.

In a study in 2007 it was found that the rate of infection of hepatitis B virus is progressing alarmingly in Pakistan (Alam et al., 2007). A total of 1300 individuals were tested for HBV infection antigens including HBsAg, anti-HBsAg, HBeAg and anti-HBcAg. A comparison of these indicators was made with different characteristics of the participants for example residential area, age, gender and social and economic characteristics to make analysis weather there is any inter relation between these variables and disease state of the patient. A total of 52 were tested positive for HBsAg. 9.30%, 33.47% and 12% individuals had HBeAg, antibodies for HBsAg, and antibodies for HBcAg respectively (Alam et al., 2007. HBsAg seropositivity rate was significantly associated (p = 0.03) with the residing locality indicating high infection in rural areas (Alam et al., 2007. Antibodies titer against HBsAg decreased with the increasing age reflecting an inverse correlation (Alam et al., 2007). These results indicate high prevalence rate of Hepatitis B virus infection and nationwide vaccination campaigns along with public awareness and educational programs are needed to be practiced urgently (Alam et al., 2007).

In a recent study in 2007 conducted by Baig et al., it was found that HBV genotyping is generally used in order to find the relationship between various virus strains and origin of infection mostly in research studies (Baig et al., 2007). In the study 295 patients who were tested positive for HBsAg from the Pakistan Medical Research Council's (PMRC) outpatient clinics were included in the research study. There were 226 male while 69 were females. Amongst the total 295 subjects, 156 (53.2%) were having Acute, 71 (24.2%) were HBV Carriers. There were 54 (18.4%) who had Chronic hepatic disease. It was found that 14 of the total 226 had e Cirrhosis of the liver.

Chapter 3


The study was conducted at Al-Rehman Medical Laboratory from February to April 2011.The number of samples were 200 for this study. The data was collected on a structured proforma (Appendix A). Written consent for each participant and brief clinical history was recorded. Three ml of blood sample was collected in a disposable BD syringe. After clotting, Serum was separated in a clear sterilized container for testing. The initial testing was carried out by Immunochromatograhic Technique (Acon, USA) and confirmed by 3rd Generation ELISA Technique (EASE BN-96 TMB, Taiwan).



Principle of immunochromatography (ICT) is the same as ELISA method, the chromatographic paper is used for the detection of immunological reactions. For this system, two kinds of specific antibodies against antigen are used. The antibodies is impregnated on the chromatographic paper, the other one is called colloidal gold which is the infiltration into a pad of samples. The sample pad is attached at the end of the membrane for the completion of immunochromatography unit. When the serum is added to the sample pad, the immunocomplex is formed by the antigen and antibody reaction. The antigen found in the sample and the antibody in the colloidal gold. Due to the chromatographic phenomenon the complex move forward and association is made with the antibody which is already present on the membrane, thus the immuno- complex is formed with the immobilized antibody, which results to form a coloured red purple line.

The red purple line shows the antigen presence, which is an indication of antigen appearance. The reaction takes the time up to 15 minutes.


For the test purpose the device known as (Acon, USA) are used, the following technique was followed. Before the assay procedure, all test reagents and the collected samples were kept to the room temperature. The test strip uncovers from the foil pouch and brings it to the dry surface. After adding three drops of patient's samples, the colour changes in the form of band were interpreted with in 15 minutes.

Interpretation of Results


The purplish red colour on the test band and the control band shows the purplish red band also, the appetence of both the bands indicate the positive test.


The appearance of puplish red band only on the control chamber indicates the negative test.



The General Biological EASE BN-96 for HBsAg assay is based on Sandwich principle (antibody - * antigen - * antibody).


Before the procedure the test reagents and the patients samples were being kept to the room temperature. room temperature also denoted as the temperature from(20-30 °C).

Three wells coated with anti-HBs were taken and 100 µl of each positive control, 100µl negative control and 100µl sample were dispensed to into appropriate well respectively. The adhesive slip was applied to each well.

The wells containing samples and controls were incubated at 37 °C in incubator or in water bath for one hour.

After incubation, the adhesive slip was removed from wells and washed 3-5 times.

100 µl of Anti-HBs peroxidase solution into each reaction well except blank.

The adhesive slip was applied again to each well.

The wells containing samples and controls were incubated at 37 °C in incubator or in water bath for one hour.

The step No. 4 was repeated.

50 µl of colour solution A was added and then 50 µl of colour solution B was added into each well.

Then all the wells were kept in the dark at room temperature for 30 minutes after applying adhesive plastic slip.

Stop solution of 100 µl was added to stop the reaction.

The absorbance of controls and test specimens was determined within 15 minutes with a spectrophotometer.

The intensity of the yellow colour shows the severity of the antigens present in the patient sample, higher the colour show the high intensity of antigens in the sample. Wells containing samples negative for HBsAg remain colorless.

Interpretation of Results


The cutoff index value for Hepatitis B is 2.0, greater the value from 2.0 indicates the positive test (reactive).


The samples show the cutoff index value lower than 2.0 shows that the sample is negative for hepatitis B surface antigen.

Chapter 4


In this study 200 blood samples were tested for Hepatitis B Virus over a period of three months in Al-Rehman Medical Laboratory Thall. Among 200 blood samples, 07 (3.5%) were the positive cases for Hepatitis B surface antigen (HBsAg); (Table 4.1 and Figure 4.1).

Table 4.1: ALL Hepatitis B Positive Cases


Total Samples

Total Number of Negative Cases

Total Number of Positive Cases

Positive Percentage






Figure 4.1: Total Number of Hepatitis B Positive Cases

These 07 cases had different histories of dental treatment (42.85%), traveled abroad (28.57%), surgery (14.28%) and blood transfusion (14.28%); (Table 4.2 and Figure 4.2).

Table 4.2: The Estimated Percentage of Different Risks Factors among General Population for HBV

S. No.

Risk Factors

Number of Positive Cases

Percentage (%)


Through Dental Treatment




Travel Abroad








Blood Transfusion



Figure 4.2: Percentage of Different Risks Factors among General Population for HBV

Out of these 07 (3.5%) cases, one sample was negative on Immunochromatography Technique (ICT) having the absorbance value of 3.0, while all 07 samples were positive on 3rd Generation ELISA (Table 4.3 and Figure 4.3).

Table 4.3: Main Comparison of Result between ICT And ELISA

Total Positive cases

Positive Cases on ELISA

Total Positive Cases on ICT




Figure 4.3: Results Comparison of ELISA and ICT

Chapter 5


There are a number of research studies conducted in Pakistan in order to find out the prevalence rate of hepatitis B and other issues related to the epidemiology of the study. The present study is conducted among the general population. The sample was selected randomly.

Pakistan is a low endemic region for hepatitis B virus as reported by WHO. The prevalence rate of hepatitis B infection is 3% of the total infections (WHO, 2008).

In 1998 in a research conducted by Hussain et al., it was found that the incidence rate of hepatitis B in Pakistan was 8% with male to female ratio of 7:1.

In Islamabad in 2002 a study was conducted at AFIT, according to which it was concluded that t 3.3% blood donors from Northern Pakistan were HBsAg positive (Khattak et al., 2002).

In another conducted in 1999 Lahore, the prevalence of HBV was found to be 2.04% in 2.06% in healthy blood donors of Faisalabad (Hashimi et al., 1999). The frequency of hepatitis B antigen determined in healthy subjects and patients with liver disease was 2.9% while 33% patients with acute viral hepatitis, 20% with cirrhosis and 10% with hepatocellular carcinoma (HCC) were HBsAg positive (Zuberi et al., 1978). HBV prevalence rate of 2% had been reported in the male volunteer blood donors of Karachi (Akhtar et al., 2005). 55% of the chronic liver disease and hepatocellular carcinoma patients were positive for HBsAg (Tong et al., 1996).

In a study conducted recently in 2008 it was found that the major risk factors involved in an outbreak in District Larkana (Sindh province of Pakistan) were intravenous drug usage. The limitations of our study include the lack of information about HBV associated risk factors like multiple blood transfusions, surgical operations, dental procedures, unsafe sex practices and frequent barber visiting.

In this study, out of 07 (3.5%) positive cases, one sample was negative (Table 4.3) on Immunochromatographic Technique (ICT) having the absorbance value of 3.0, while all the 07 samples were positive on 3rd Generation ELISA Method. The reason might be the lower sensitivity of the screening test. ELISA method can detect small quantity of HBsAg in the serum due to high sensitivity, while ICT only detects HBsAg in the serum when present in large quantity. These observations clearly indicate the serious problem and raise many questions on the sensitivity of the test. Lower sensitivity of the test can give false negative result. Immunochromatographic Technique (ICT) missed one sample out of 07 and gave false negative result. It was observed in this study that ELISA Method is more specific than ICT for routine blood donors screening.

The similar observation has been made by Abdelbagi et al. 2007. He reported that ELISA method is more specific and sensitive than ICT method. Khan et al. 2010 and Mustafa et al. 2009 reported the same results by using ELISA Method. They found that the ELISA method is more specific and sensitive than ICT for screening blood tests.

Chapter 6



In our country Hepatitis B virus is very much endemic. Too many liver diseases may be caused by Hepatitis B virus for example, acute hepatitis, chronic hepatitis, fulminant hepatic failure and may even advance to carrier state

.The purpose of this study was to identify HBV and its risks factors in the general population of Thall. This study was conducted at Al-Rehman Medical Laboratory from February to April 2011.

A total of 200 blood samples were tested for HBV. All blood samples were initially tested by using Immunochromatograhic Technique (ICT) and confirmed by using 3rd Generation Enzyme Immunosorbent Assay (ELISA). The results of ICT and ELISA were also compared. Through this study it was found that seven cases were found positive for hepatitis B surface antigen out of the total 200 blood samples,

These 07 cases had different histories of dental treatment (42.85%), traveled abroad (28.57%), surgery (14.28%) and blood transfusion. Out of these 07 cases, one sample was negative on Immunochromatographic Technique (ICT), while all 07 the samples were positive on ELISA 3rd Generation Method. From this study it is concluded that ELISA method is superior as well as very much reliable as compared to immunochromatography technique.


The number of positive cases of Hepatitis B virus were 07 (3.5%) in 200 blood samples collected from different areas of Thall.

It can be concluded that 3rd Generation ELISA is more specific and sensitive than ICT and shows good results.

Dental treatment shows(42.85%) which is the main source for the transmission of HBV infection.


The above results of the study shows that the diagnosis of Hepatitis B on ELISA is more reliable technique to detect Hbs Ag in the blood. The preventive strategies should be adopted for Hepatitis B virus which is an important issue on public health aspect.

Health educational and promotional programmes should be formulated and implemented on the country level.

Vaccination for HBV should be adopted at national level.

The true prevalence rate of Hepatitis B is not yet known in Pakistan therefore it is suggested that further sponsored studies should be conducted throughout the country.