Incidence Of Hepatitis B Virus And Risk Factors Biology Essay

Hepatitis B virus is extremely endemic in Pakistan. It may do a broad assortment of ague or chronic hepatic and extrahepatic diseases, every bit good as chronic bearer state.The intent of this survey was to place HBV and its hazards factors in the general population of Thall.


This survey was conducted at Al-Rehman Medical Laboratory from February 2011 to April 2011. A sum of 200 blood samples were tested for HBV. All blood samples were ab initio tested by utilizing Immunochromatograhic Technique ( ICT ) and confirmed by utilizing 3rd Generation Enzyme Immunosorbent Assay ( ELISA ) . The consequences of ICT and ELISA were besides compared.

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Among 200 blood samples, 07 instances were positive for Hepatitis B surface antigen ( HBsAg ) .

These 07 instances had different histories of dental intervention ( 42.85 % ) , traveled abroad ( 28.57 % ) , surgery ( 14.28 % ) and blood transfusion. Out of these 07 instances, one sample was negative on Immunochromatographic Technique ( ICT ) , while all 07 the samples were positive on ELISA 3rd Generation Method. These consequences suggested that the rapid immuno-chromatographic kit for HBsAg has merely limited efficaciousness and must be confirmed through superior method like ELISA.


The figure of positive instances of Hepatitis B virus were 07 ( 3.5 % ) in 200 blood samples collected from different countries of Thall.

It can be concluded that 3rd Generation ELISA is more specific and sensitive than ICT and shows good results.Dental intervention ( 42.85 % ) was the chief hazard factor for transmittal of HBV infection. The preventative schemes should be adopted for Hepatitis B virus which constitutes a major public wellness job in Pakistan.

Abbreviations and Acronyms

HBV Hepatitis B VirusHCV Hepatitis C VirusRNA Ribo Nucleic AcidELISA Enzyme Linked Immunosorbent AssayMEIA Microparticle Enzyme ImmunoassayALT Alanine AminotransfrasePCR Polymerase Chain ReactionWHO World Health OrganizationCRF Chronic Renal FailureTTVI Transfusion-Transmissible Viral InfectionDeoxyribonucleic acid Deoxyribo Nucleic AcidOR Odds RatiosHBsAg Hepatitis B surface antigenHBeAg Hepatitis B vitamin E antigenAnti-HBe Anti- Hepatitis B vitamin E antigenHBcAg Hepatitis B nucleus antigenEIA Enzyme immunoassayRIBA Recombinant Immunoblot AssayICT Immunochromatographic Techniquemilliliter Mili litresAµl Micro litresTMB T Etramethyl-BenzidineNAT Nucleic acid provingN NumberIgG Immunoglobulin GA°C Centigrade


Certificate of Approval IDeclaration twoAcknowledgement fourIntroduction VMethods VConsequences vDecision sixAbbreviations and Acronyms sevenContentss viiiChapter 1 1Introduction 1Chapter 2 4LITERATURE REVIEW 4Chapter 3 14METHODOLOGY 14IMMUNOCHROMATOGRAPHY TECHNIQUE 14Enzyme-linked-immunosorbent serologic assay FOR THE DETECTION OF HBsAg 16Chapter 4 18RESULTS 18Chapter 5 22DISCUSSION 22Chapter 6 25SUMMARY, CONCLUSION AND RECOMMENDATIONS 25SUMMARY 25Decision 26RECOMMENDATIONS 26Mentions 27APPENDIX-A 32Questionnaire 32APPENDIX-B 33Consent signifier for the participants 33

Chapter 1


Hepatitis B is found worldwide but is peculiarly prevailing in Asia, globally about 350 million people are inveterate infected with HBV and approximately 75 % of them are Asiatic. Hepatitis B virus causes hepatitis infection which is complex construction two-base hit stranded DNA virus. The envelope contains a protein called the surface antigen ( HBsAg ) .

There are two other of import antigens, which are the nucleus antigen and the vitamin E antigen.The nucleus antigen is known as ( HBcAg ) and the vitamin E antigen is known as ( HBeAg ) . HbeAg is an of import index of transmissibility because the vitamin E antigen is located on the nucleus protein, which means the infective atom must be present in the patient ‘s blood ( Warren Levinson, 2004 ) .The prevalence of hepatitis B virus have different scope in different countries of the universe ( Lavanchy, 2004 ) , In the in-between E, sub-Saharan Africa and far east the prevalence show the high rate runing to greater than 8 % positiveness while in the developed states like Northern Europe, United States and in Australia the ratio is intermediate which is runing from 2 % to 7 % . In some other states like India and Japan and the countries of cardinal Asia ( Andre,2000 ) .Study shows that prevalence rate in Pakistan is 35 % to 38 % in which 4 % are count as the bearer ( 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 wellness attention personals have 2.4 % prevalence rate in Karachi wellness ( Khichi and Channar, 2000 ) , s In Abbottabad the prevalence rate was 2.

68 % in 5,207 givers ( Ally et al. , 2005 ) . In Pakistan 903 afghan refugees were screened holding 8.3 % prevalence rate ( Quddus et al. , 2006 ) ) .

Harmonizing to Zuberi ( 1996 ) the 10 % of healthy grownups were found the HBV bearers.There are three chief manners of transmittal, via sexual intercourse, parentally from female parent to new born and blood transfusion. It is concluded that the 5 % of patients with HBV infection become chronic bearer i.e. person who has HBsAg prevailing in their blood for at least six months ( Warren Levinson, 2004 ) .

Vertical transmittal from infected female parent to their new born kid is of great importance. Women ‘s those holding HBs Ag bearers can convey the virus ; nevertheless, but the grade of the infection is different from single to single. The transmittal of HbeAg is greater in septic female parents as compared with HBe. HBV infection in babyhood plays a cardinal function which leads to chronicity. The babies with 2 to 3 months, goes to the bearer phase with the per centum runing from 80 % to 85 % ( Kazmi et al. , 2003 ) .Hazard factors by which the Hep B spread are the direct contact with Hep B infected individual, insecure sexual contacts, through endovenous drug utilizations, through different surgical and dental processs ( Sali et al.

, 2005 ) . The chronic bearer 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 of import public wellness issue in blood givers with acute hepatitis rate of 6 % . About 1 % of the populations are symptomless chronic HBsAg bearer, while 39 % of patients enduring from chronic liver disease and 37 % with hepatocelluler carcinoma are HBsAg seropositive ( Karki et al. , 2008 ) .In the same scenario, this survey has been designed to place HBV, acute infection ( active ) and chronic infection ( bearer province ) in blood givers in our local environment.


To place Hepatitis B in blood givers.To happen out the active infection and bearer province in blood givers who are the beginning of infection for others.To compare the consequences of ICT and ELISA Method for designation of HBV.

Chapter 2


Khan et al. 2010 conducted a survey at the King Edward Medical University, Lahore during the period between November 2009 and January 2010. Four rapid immuno-chromatographic checks – ONE STEP HBsAg One Check, One measure HBsAg trial Accurate were evaluated for sensing of hepatitis B surface antigen.

Sum of 57 tested for Hepatitis B by ELISA were stored at -20 A°C and later tested with rapid ICT trials ( devices ) ( Accurate and One Check trade names ) . Sensitivity, specificity, positive prognostic value, negative prognostic value and diagnostic truth of these trials were calculated utilizing ELISA as the gilded criterion. This survey was conducted to measure the public presentation 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 cheque and 19 samples were positive by accurate. The sensitiveness for one cheque was found to be 53 % and 50 % for accurate. The negative prognostic value for one cheque was 51 % and 49 % for Accurate. The positive predictive values for HBV were 100 % for one cheque and 95 % with Accurate.Ali et Al.

2009 evaluated that Pakistan carries one of the universe ‘s highest loads of chronic hepatitis and mortality due to liver failure and hepatocellular carcinomas. However, national degree estimations prevalence and hazard factors for hepatitis B. They observed the medical and public wellness literature over a 13-year period ( 1994-2007 ) to measure the prevalence of active hepatitis B and chronic hepatitis C in Pakistan. They analyzed informations individually for the general and bad populations for each of the four states.

A leaden norm of hepatitis B antigen prevalence among healthy grownups ( blood givers 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 countries of Pakistan. The published literature on the manners of transmittal of hepatitis B in Pakistan implicate contaminated needle usage in medical attention and drug maltreatment and insecure blood and blood merchandise 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 universe. It is estimated that worldwide more than two billion people infected by HBV and 350 million people have chronic infection.

The HBV is extremely contagious and transmittal occurs through transdermal or permucosal paths, and morbific blood or organic structure fluids, through sexual contact or by contaminated acerate leafs. In sub-Saharan Africa, transfusion-transmitted HBV infection is progressively going a major manner of transmittal of HBV in the high-prevalence countries. The demand for blood transfusion services is high and increase the possibility of the transmittal of HBV through contaminated blood Due to endemicity of infections doing anaemia, malnutrition, and surgical and obstetrical exigencies associated with blood loss in the sub-Saharan Africa. Blood safety remains a large issue in transfusion medical specialty in this portion of the universe because national blood transfusion services and policies, appropriate substructure, trained forces and fiscal resources are unequal. As portion of public wellness interventional steps, the transmittal of HBV can be minimized by pre-screening contribution, by rating of bad givers.Alavian et Al. 2008 observed that Hepatitis B virus ( HBV ) infection is a planetary job. It is estimated that 400 million people are enduring from this infection.

They reviewed to set all grounds on HBV infection in Iran and to do an accurate estimation of HBV infection prevalence in Iran for farther planning to command the infection. Meta-analysis and study informations analysis of all national and international documents, theses, Congresss, studies, Persian medical universities undertakings, research centres, studies of Deputy for Health personal businesss ( published or unpublished ) Persian 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 aims, proper trying method with indistinguishable and valid measuring instruments for all survey topics and proper analysis methods sing trying design and demographic accommodations Presence of positive HBsAg in blood samples of survey samples Fourteen surveies met the inclusion standards. They were from 7 ( out of 30 ) states in which about 40 per centum of the state population live. These states ( 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 per centum, in work forces and adult females 2.55 per centum and 2.03 per centum severally.

About 1.5 million people in Iran are populating with HBV infection and it is estimated that 15 % to 40 % of them are at hazard of developing cirrhosis and/or hepatocellular carcinoma ( HCC ) without intercession. The prevalence of HBV infection has been reported higher in more recent surveies compared to the survey in 2000-2001.

Behal et Al. 2008 determined that Infection with hepatitis-B virus has been a important cause of morbidity claiming more than a million lives every twelvemonth. Epidemiological information reveals that there are 360 million bearers of hepatitis-B virus throughout the Earth and 78 % of the universe populations ‘ hail from Asia. Though several surveies from Indian sub-continent have provided an estimation of the prevalence of this viral infection, there exist merely few surveies, which reflect the position in the general population. The present survey was conducted to look into the prevalence of hepatitis-B infection in North Indian general population. A sum of 20,000 healthy blood givers were screened for hepatitis-B surface antigen ( HBsAg ) position utilizing 3rd coevals ELISA kit.

Seroprevalence rate of seropositive givers was calculated and classified by age, sex and blood groups. Statistical analysis was done by utilizing trials of proportions, chi-square and assurance interval. Out of 20,000 givers, 450 ( 2.25 % ) were HBsAg positive ( 95 % assurance 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 survey in the new campus of Punjab University Lahore. In the survey they checked the pupils 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 % severally. it is indispensable that all givers should be screened for anti- HCV and HbsAg in order to forestall the transmittal of HBV and HCV through blood transfusion. It is necessary to wake up the consciousness about it.It was observed in 2007 by Alavian et Al. that during the last decennary, the prevalence of Hepatitis B virus ( HBV ) has decreased in Persian population. The ground for this lessening in the prevalence of Hepatitis B is likely the addition in consciousness about HBV hazard factors, national inoculation plan since 1993 for all newborns, and inoculation of high hazard groups. Hepatitis B inoculation was included in the EPI in 1993 and after 13 old ages of its execution, the coverage has reached well from 62 % in 1993 to 94 % in 2005. With immunisation of high hazard people for Hepatitis B virus, surveillance of persons infected with hepatitis B and control on the wellness position of refugees will farther take down the rate of Hepatitis B infection in Iran.

It is really of import to command all the possible paths of transmittal of Hepatitis B virus in order to command its spread. Practice of Universal safeguards in the wellness establishments plays a cardinal function in the spread of Hepatitis B and other unsafe viral infections.In 2007, it was reported by Cohen, that certain viruses called hepatotropic virus inflame the live of the topic, which is diffuse in nature and is called acute viral hepatitis. Other than nonspecific symptoms of viral diseases, there can be nausea, purging and even loss of appetency. Sometimes it is associated with febrility every bit good as hurting in the venters peculiarly in the right upper quarter-circle. Subsequently on when other marks and symptoms disappear, xanthous stain of the tegument and sclerotic coat of the eyes appear which is called icterus. Majority of the instance decide but in rare instances, chronic hepatitis develop.Hepatitis B is diagnosed by the designation of virus in the organic structure, via different trials on blood.

Far bar of the viral hepatitis, personal hygiene is really of import. Pre exposure and station exposure prophylaxis may be possible utilizing vaccinums or serum globulins depending on specific virus. There is no specific intervention, merely supportive steps are taken and symptoms are treated consequently.Globally acute viral hepatitis is really common. There is assortment of causes of this viral infection.In 2007, Essa and Farhan, 2007 pointed out that viral hepatitis is chief job in Pakistan and HBV is extremely antigenic. In this survey a sum of 35,257 topics of including males every bit good females were screened, where HBsAg has to be detected.

The survey lasted for eight old ages from which ended in 2006.In many Asiatic states Hepatitis B virus is endemic as observed by Liuac et Al. in 2006.

Transfusion is the most common path among many transmittal paths that should be prevented. In early 1970s, hepatitis B surface antigen was implemented, which was no uncertainty a great success in heightening the safety of transfusion. However, it was demonstrated that transmittal of the said virus by blood negative for HBsAg can still be possible in the early phases of infection or during chronic phases of infection ( i.e.

“ supernatural ” 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 restrictions in current blood showing patterns, OHB is an unmarked beginning of HBV transmittal.

New policy for testing should be formulated and imposed on the footing of available informations or freshly designed surveies.In order to mensurate the degree of HBsAg on ELISA technique, a survey was carried out in Jordan. The said survey was conducted in 1997 on 247 kids who did non acquire their inoculation. I in 2004 247 more childs, who had their inoculation done, were studied.

A sum of 8 individuals are HBs-Ag positive and merely one kid ( 0.4 ) is positive from the vaccinated group.In 2003, Bhatta et Al. performed a survey in Khatmandu in a third attention infirmary.

The entire continuance of survey was nine months from April to December. The aim was to mensurate the prevalence of Hepatitis B in the patients coming to the instruction infirmary for intervention intents. 200 patients participated in the survey. Blood from the participants was collected and tested for the presence of Hepatitis B virus.

Amongst all, entire 5 were tested positive all of whom were male of immature age.In 2003, in a survey the research worker Roger et Al. obtained epidemiologic and serologic informations on 422 family contacts of 157 blood giver campaigners in the Baltimore Regional Red Cross Blood Program who were symptomless chronic bearers of hepatitis B surface antigen ( HBsAg ) ( index bearers ) , and on 318 family contacts of 157 blood givers who were negative for serologic markers of hepatitis B virus ( index controls ) . Index bearers were matched to index controls and their family contacts were similar with regard to race, sex, and instruction degree completed.

HBsAg prevalence was 6.8 times higher, and antibody to HBsAg ( anti-HBs ) prevalence 2.7 times higher among the contacts of bearers. The combined prevalence for hepatitis B virus markers was 29.9 % among the contacts of bearers compared to 8.

8 % among the contacts of controls ( P & lt ; 0.001 ) . Bing colored, the sibling of a bearer, of older age, of lower educational position, and a member of a little family were significantly associated with either a higher antibody or combined hepatitis B prevalence rate ( Roger et al. 2003 ) .The contacts of female bearers had an HBsAg prevalence rate 4.

7 times greater than the contacts of male bearers ( Roger et al. 2003 ) . Children who had a female parent as their index bearer had an 11 crease higher HBsAg prevalence than did kids who had a male parent as their index bearer ( 18.0 % vs. 1.6 % , P & lt ; 0.005 ) ( Roger et al.

2003 ) . Epidemiologic features of the contacts or their Index bearers, such as the sharing of Items belonging to the index bearer, the type and frequence of sexual dealingss, and the frequence of possible exposure to organic structure fluids of the index bearers were examined. Contacts of bearers who had had skin lesions in the past twelvemonth had a higher prevalence of serologic markers for hepatitis B than contacts of bearers without such lesions. ( Roger et al.

2003 ) . No sex-related factors among partners were significantly associated with an increased prevalence of serologic markers ( Roger et al. 2003 ) .

These informations give suggest that person-to-person manners of transmittal are of import in the spread of hepatitis B within families. The forms of hepatitis B serologic markers observed here suggest that female parent to child transmittal is most of import in accounting for the high HBsAg prevalence among the contacts ; other soon unidentified but close types of contact best explain the forms of antibody prevalence.Hussain et al conducted a survey in 1998 in Pakistan.

Harmonizing to his survey, Hepatitis B virus ( HBV ) is really much endemic in Pakistan, particularly in rural countries. It can be the cause of all types of Hepatitis, like acute hepatitis, chronic hepatitis. It may besides ensue in fulminate hepatic failure or may even take the signifier into bearer province.Franchis et Al. found in 1993 that those transporting surface antigen of hepatitis B with normal liver map trials, took into history the carcinoma of hepatic cells and other serious unrecorded diseases.

Majority ( 69 ) of the participants had normal findings on the footing of histology while 18 persons had chronic hepatitis of serious nature and 5 were holding mild disease of the unrecorded. Most the participants had Serum enzyme degrees within normal bounds which was 58 of 68 participants who had regular followup. Out of the all 68 participants, 3 had chemical alterations which were associated to hepatitis B virus ( HBV ) infection ( Franchis et al. ) In one of these patients, a ulterior histologic rating showed patterned advance 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 non hold regular followup, except 1, were interviewed by telephone during 1990 ( Franchis et al. ) .

All of them were accepting that they had liver disease of any sort. None of the topics had carcinoma of hepatic cells.In a survey in 2007 it was found that the rate of infection of hepatitis B virus is come oning alarmingly in Pakistan ( Alam et al. , 2007 ) .

A sum of 1300 persons were tested for HBV infection antigens including HBsAg, anti-HBsAg, HBeAg and anti-HBcAg. A comparing of these indexs was made with different features of the participants for illustration residential country, age, gender and societal and economic features to do analysis conditions there is any inter relation between these variables and disease province of the patient. A sum of 52 were tested positive for HBsAg. 9.30 % , 33.47 % and 12 % persons had HBeAg, antibodies for HBsAg, and antibodies for HBcAg severally ( Alam et al.

, 2007. HBsAg seropositivity rate was significantly associated ( p = 0.03 ) with the shacking vicinity bespeaking high infection in rural countries ( Alam et al. , 2007. Antibodies titre against HBsAg decreased with the increasing age reflecting an reverse correlativity ( Alam et al.

, 2007 ) . These consequences indicate high prevalence rate of Hepatitis B virus infection and countrywide inoculation runs along with public consciousness and educational plans are needed to be practiced desperately ( Alam et al. , 2007 ) .In a recent survey in 2007 conducted by Baig et al. , it was found that HBV genotyping is by and large used in order to happen the relationship between assorted virus strains and beginning of infection largely in research surveies ( Baig et al. , 2007 ) .

In the survey 295 patients who were tested positive for HBsAg from the Pakistan Medical Research Council ‘s ( PMRC ) outpatient clinics were included in the research survey. There were 226 male while 69 were females. Amongst the entire 295 topics, 156 ( 53.2 % ) were holding Acute, 71 ( 24.2 % ) were HBV Carriers. There were 54 ( 18.4 % ) who had Chronic hepatic disease.

It was found that 14 of the entire 226 had e Cirrhosis of the liver.

Chapter 3


The survey was conducted at Al-Rehman Medical Laboratory from February to April 2011.The figure of samples were 200 for this survey. The information was collected on a structured proforma ( Appendix A ) . Written consent for each participant and brief clinical history was recorded.

Three milliliter of blood sample was collected in a disposable BD syringe. After coagulating, Serum was separated in a clear sterilised container for proving. 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 sensing of immunological reactions. For this system, two sorts of specific antibodies against antigen are used. The antibodies is impregnated on the chromatographic paper, the other 1 is called colloidal gold which is the infiltration into a tablet of samples. The sample tablet is attached at the terminal of the membrane for the completion of immunochromatography unit. When the serum is added to the sample tablet, 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 frontward and association is made with the antibody which is already present on the membrane, therefore the immuno- composite is formed with the immobilized antibody, which consequences to organize a colored ruddy purple line.The ruddy purple line shows the antigen presence, which is an indicant of antigen visual aspect. The reaction takes the clip up to 15 proceedingss.


For the trial purpose the device known as ( Acon, USA ) are used, the undermentioned technique was followed. Before the assay process, all trial reagents and the gathered samples were kept to the room temperature. The trial strip uncovers from the foil pouch and brings it to the dry surface.

After adding three beads of patient ‘s samples, the coloring material alterations in the signifier of set were interpreted with in 15 proceedingss.

Interpretation of Consequences


The purple ruddy coloring material on the trial set and the control set shows the purple ruddy set besides, the appetite of both the sets indicate the positive trial.


The visual aspect of puplish ruddy set merely on the control chamber indicates the negative trial.

Enzyme-linked-immunosorbent serologic assay FOR THE DETECTION OF HBsAg


The General Biological EASE BN-96 for HBsAg check is based on Sandwich rule ( antibody – * antigen – * antibody ) .


Before the process the trial reagents and the patients samples were being kept to the room temperature. room temperature besides denoted as the temperature from ( 20-30 A°C ) .

Three Wellss coated with anti-HBs were taken and 100 Aµl of each positive control, 100Aµl negative control and 100Aµl sample were dispensed to into appropriate good severally. The adhesive faux pas was applied to each well.The Wellss incorporating samples and controls were incubated at 37 A°C in brooder or in H2O bath for one hr.After incubation, the adhesive faux pas was removed from Wellss and washed 3-5 times.100 Aµl of Anti-HBs peroxidase solution into each reaction good except space.The adhesive faux pas was applied once more to each well.The Wellss incorporating samples and controls were incubated at 37 A°C in brooder or in H2O bath for one hr.The measure No.

4 was repeated.50 Aµl of color solution A was added and so 50 Aµl of color solution B was added into each well.Then all the Wellss were kept in the dark at room temperature for 30 proceedingss after using adhesive plastic faux pas.Stop solution of 100 Aµl was added to halt the reaction.The optical density of controls and trial specimens was determined within 15 proceedingss with a spectrophotometer.The strength of the xanthous coloring material shows the badness of the antigens nowadays in the patient sample, higher the coloring material show the high strength of antigens in the sample. Wells incorporating samples negative for HBsAg remain colorless.

Interpretation of Consequences


The cutoff index value for Hepatitis B is 2.

0, greater the value from 2.0 indicates the positive trial ( 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 survey 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 instances for Hepatitis B surface antigen ( HBsAg ) ; ( Table 4.1 and Figure 4.1 ) .

Table 4.1: ALL Hepatitis B Positive Cases


Entire SamplesEntire Number of Negative CasesEntire Number of Positive CasesPositive PercentageHBsAg200193073.5 %

Figure 4.

1: Entire Number of Hepatitis B Positive Cases

These 07 instances had different histories of dental intervention ( 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.

Hazard 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 % ) instances, one sample was negative on Immunochromatography Technique ( ICT ) holding the optical density 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

Entire Positive instancesPositive Cases on ELISA

Entire Positive Cases on ICT


Figure 4.

3: Consequences Comparison of ELISA and ICT

Chapter 5


There are a figure of research surveies conducted in Pakistan in order to happen out the prevalence rate of hepatitis B and other issues related to the epidemiology of the survey. The present survey is conducted among the general population. The sample was selected indiscriminately.Pakistan is a low endemic part for hepatitis B virus as reported by WHO. The prevalence rate of hepatitis B infection is 3 % of the entire 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 survey was conducted at AFIT, harmonizing to which it was concluded that t 3.

3 % blood givers 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 givers of Faisalabad ( Hashimi et al.

, 1999 ) . The frequence of hepatitis B antigen determined in healthy topics 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 voluntary blood givers 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 survey conducted late in 2008 it was found that the major hazard factors involved in an eruption in District Larkana ( Sindh state of Pakistan ) were endovenous drug use. The restrictions of our survey include the deficiency of information about HBV associated hazard factors like multiple blood transfusions, surgical operations, alveolar consonant processs, insecure sex patterns and frequent Barber visiting.In this survey, out of 07 ( 3.5 % ) positive instances, one sample was negative ( Table 4.

3 ) on Immunochromatographic Technique ( ICT ) holding the optical density value of 3.0, while all the 07 samples were positive on 3rd Generation ELISA Method. The ground might be the lower sensitiveness of the screening trial. ELISA method can observe little measure of HBsAg in the serum due to high sensitiveness, while ICT merely detects HBsAg in the serum when nowadays in big measure. These observations clearly indicate the serious job and raise many inquiries on the sensitiveness of the trial. Lower sensitiveness of the trial can give false negative consequence. Immunochromatographic Technique ( ICT ) missed one sample out of 07 and gave false negative consequence.

It was observed in this survey that ELISA Method is more specific than ICT for everyday blood givers testing.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 consequences by utilizing ELISA Method. They found that the ELISA method is more specific and sensitive than ICT for testing blood trials.

Chapter 6



In our state Hepatitis B virus is really much endemic. Too many liver diseases may be caused by Hepatitis B virus for illustration, acute hepatitis, chronic hepatitis, fulminant hepatic failure and may even progress to bearer province.The intent of this survey was to place HBV and its hazards factors in the general population of Thall.

This survey was conducted at Al-Rehman Medical Laboratory from February to April 2011.A sum of 200 blood samples were tested for HBV. All blood samples were ab initio tested by utilizing Immunochromatograhic Technique ( ICT ) and confirmed by utilizing 3rd Generation Enzyme Immunosorbent Assay ( ELISA ) . The consequences of ICT and ELISA were besides compared. Through this survey it was found that seven instances were found positive for hepatitis B surface antigen out of the entire 200 blood samples,These 07 instances had different histories of dental intervention ( 42.85 % ) , traveled abroad ( 28.57 % ) , surgery ( 14.28 % ) and blood transfusion.

Out of these 07 instances, one sample was negative on Immunochromatographic Technique ( ICT ) , while all 07 the samples were positive on ELISA 3rd Generation Method. From this survey it is concluded that ELISA method is superior every bit good as really much reliable as compared to immunochromatography technique.


The figure of positive instances of Hepatitis B virus were 07 ( 3.5 % ) in 200 blood samples collected from different countries of Thall.It can be concluded that 3rd Generation ELISA is more specific and sensitive than ICT and shows good consequences.

Dental intervention shows ( 42.85 % ) which is the chief beginning for the transmittal of HBV infection.


The above consequences of the survey shows that the diagnosing of Hepatitis B on ELISA is more dependable technique to observe Hbs Ag in the blood. The preventative schemes should be adopted for Hepatitis B virus which is an of import issue on public wellness facet.Health educational and promotional programmes should be formulated and implemented on the state degree.Inoculation for HBV should be adopted at national degree.The true prevalence rate of Hepatitis B is non yet known in Pakistan therefore it is suggested that farther sponsored surveies should be conducted throughout the state.


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