Relation To Blood Groups And Rhesus Biology Essay

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To study the frequency of Hcv genotype and their association with blood, which blood group is more Hcv positive and genotyping of that blood group to evaluate which genotype is comparatively high.

Introduction: Hepatitis C Virus (HCV) was isolated in 1989; is a major pathogen of hepatitis, An estimated 200 million people worldwide are infected with hepatitis C worldwide including approximately 17 million in Pakistan . Hepatitis C virus (HCV) is a small (55-65 nm in size), enveloped, positive sense single strand RNA virus in the family Flaviviredea. It is estimated that 170 million people are chronically infected with HCV and 3 - 4 million is newly infected .Each year. Overall frequency of HCV infection in general population of Pakistan ranges from 4 - 25% as shown by different studies. It is well known that incidence of certain diseases is related to blood group type. For example, type O "non-secretors" have about twice the incidence of duodenal ulcer than secretors of types A and B. On the other hand, type A carries a higher incidence of tumours of salivary glands, stomach and pancreas than do type O blood groups. Similarly individuals who lack Duffy system antigen are protected against infection by Plasmodium vivax. In Pakistan, it has been observed that frequency of blood group B was significantly higher and that of blood group O was lower in Diabetes mellitus type 2. . However, no study from Pakistan could be found showing distribution of hepatitis c (HCV) & genotyping- their relation to blood groups and rhesus (Rh) factor.

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Present study was carried out studies on the distribution of hepatitis c (HCV) & genotyping- their relation to blood groups and rhesus (Rh) factor in institute of public health.

Materials and Methods:

The data for the present study was collected from eleven thousand five hundred eighteen (11518) donors who came in blood bank for donate blood. These are screened for HCV antibodies. (Anti-Hcv) by rapid test devices based on immuno-chromatographic technique following the instruction given by the manufacturer

In the present study, devices manufactured by Acon, USA were used. The specimens

Reactive on screening by devices were confirmed on ELISA. Those samples are selected which are highest prevalence of Hcv infection was found in individuals with blood group B and Rh positive.

RNA is extracted from these samples using Qiagin kit. RNA is amplified by Real Time PCR. These amplified products are processed for Genotypes by Real Time Technique by using INVADER HCV Genotyping Assay. The results were subjected to chi-square. Analysis for determination of statistical variance between the values of different categories.

Result: The data result showed that there is high prevalence of 3a genotype among the patients with blood group B and Rh positive at I.P.H Lahore. The percentage of 3a was 80%, 1a is 18% and 3b is detected in this study.

Key word: HCV, RNA, PCR, HCC, EDTA, RT-PCR, Genotype.

CHAPTER 01

INTRODUCTION

1.1 Hepatitis C Virus:

Hepatitis C virus (HCV) is a small (55-65 nm in size), enveloped, positive-sense single-stranded RNA virus of the family Flaviviredea. Hepatitis C virus is the cause of hepatitis C in humans. The hepatitis C virus belongs to the genus Hepacivirus a member of the family Flaviviredea. Until recently it was considered to be the only member of this genus. However a member of this genus has been discovered in dogs - canine hepacivirus. (Kapoor A et al 2011). [1] 

1.2 Structure:

The hepatitis C virus particle consists of a core of genetic material (RNA), surrounded by an icosahedral protective shell of protein, and further encased in a lipid (fatty) envelope of cellular origin. Two viral envelope glycoproteins, E1 and E2, are embedded in the lipid envelope. (Op De Beeck A, Dubuisson J 2003). [2] 

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Figure No. 01

1.3 Transmission:

sexual activates and practice were initially identified as potential sources of exposure to the hepatitis C virus more recently studies question this route of transmission. Currently it is felt to be a means of rare transmission of hepatitis C infection .these are simple the current know modes of transmission and due to the nature of hepatitis there may be more ways that it is transmitted then the current know methods. [3] 

Injection drug use:

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Those who currently use or have used drug injection as their delivery route for drugs are at increased risk for getting hepatitis C because they may be sharing needles or other drug paraphernalia(includes cookers, cotton spoons, water etc) which may be contaminated with HCV-infected blood. An estimated 60% to 80% of intravenous recreation drug users in united state have been infected with Hcv. Harm reduction strategies are encouraged in many countries to reduce the spread of hepatitis C, through education, provision of clean needles and syringes, and safer injection technique .for that are not clear transmission by this route currently appears to be decling in USA. [4] 

Blood product:

Blood transfusion: the virus was first time isolated in 1889 and reliable test to screen for virus were not available until 1992.there fore, those who received blood or blood product prior to the implementation of screening the blood supply for Hcv may have been exposed to the virus .blood product included clotting factor and other blood product.

Iatrogenic medical or dental exposure:

People can be exposed to Hcv inadequately or improper sterilized medical or dental equipment. Equipment that may harbor contaminated blood if improperly sterilized includes needles or syringe, hemodialysis equipment, oral hygiene instruments, and jet air guns etc.

Sexual exposure:

Heterosexual or homosexual activity with multiple sexual partners has been clearly identified as a mode of transmission, but the exact risk is unknown. Because of the lack of sufficient information, persons in long-term, monogamous relationships are not advised to change sexual practices. Day-to-day contact with another household member that has hepatitis C has also been strongly implicated

Hepatitis C is believed to be transmitted only by blood. However, unlike many other blood borne viruses (like HIV) virtually any source of blood or blood products seems to be capable of carrying the virus, even if the source is indirect - like a used razor, for example. This makes hepatitis C far more transmissible than most other blood borne viruses - including HIV. Tattooing, as well as many body piercing practices, such as acupuncture and ear-piercing, have contributed significantly to the spread of HCV, even in industrial nations. Needle-stick injuries, contaminated medical equipment, and blood spills in health care settings are also responsible for many cases of HCV.

Body piercing and tattoos:

Tattoos dyes styles and piercing implement can transmit Hcv-infected blood from one person to another if proper sterilization techniques are not followed.

Shared personal care item:

Certain specialized risks have also been identified - such as manicures, shared toothbrushes, or straight razors in barber shops.

Vertical transmission:

. Maternal-infant transmission has also been documented as a mode of spread. Transmission occur among women who are Hcv Rna positive at the time of delivery; the risk of transmission in this setting is approximately 6 out of 100 .among women who are both Hcv and Hiv positive at the time of delivery the risk of transmitting Hcv ids increased to approximately 25 out 100.

Table No. 01

1.4 Replication:

Replication of HCV involves several steps. The virus replicates mainly in the hepatocytes of the liver, where it is estimated that daily each infected cell produces approximately fifty virions (virus particles) with a calculated total of one trillion virions generated. The virus may also replicate in peripheral blood mononuclear cells, potentially accounting for the high levels of immunological disorders found in chronically infected HCV patients. HCV has a wide variety of genotypes and mutates rapidly due to a high error rate on the part of the virus' RNA-dependent RNA polymerase. The mutation rate produces so many variants of the virus it is considered a quasispecies rather than a conventional virus species.

1.5 Hepatitis "C" Disease:

Hepatitis C virus (HCV) is a blood-borne disease that causes inflammation of the liver and to which there is currently no vaccine available. The World Health Organization (WHO) estimates that 3% of the world's populations, approximately 170 million people are infected with HCV and it is a leading cause of liver cirrhosis, end stage liver disease, hepatocellular carcinoma

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(HCC) and liver transplantation [5] 

Many people infected with hepatitis C have no symptoms. When symptoms are present, they can range from mild to severe. The most common early symptoms are mild fever, headache, muscle aches, fatigue, and loss of appetite, nausea, vomiting and diarrhea. Later symptoms may include dark coffee-colored rather than dark yellow urine, clay-colored stools, abdominal pain, and yellowing of the skin and/or whites of the eyes.

C:\Users\ACER\Desktop\HCV outcomes.jpg

Table No. 02

1.6 Genome:

Hepatitis C virus has a positive sense single-stranded RNA genome. The genome consists of a single open reading frame that is 9600 nucleotide bases long. This single open reading frame is translated to produce a single protein product, which is then further processed to produce smaller active proteins. At the 5' and 3' ends of the RNA are the UTR that are not translated into proteins but are important to translation and replication of the viral RNA. The 5' UTR has a ribosome binding site. (Jubin R 2001). [6] 

1.7 Global Prevalence:

An estimated 170 million people are infected with the Hepatitis C virus (HCV) worldwide, says the World Health Organization. The predominance of the Hepatitis C-Virus infection however, varies around the world. Just a decade ago, Frank et al reported that Egypt has the highest number of reported HCV infections, due to the frequent use of contaminated parenteral antischistomal therapy. The subsequent effect was a mean prevalence of HCV antibiotic in persons in Egypt, numbering up to 22%. Research estimates that 1.8% of the United States population is positive for Hepatitis C viral antibodies. This is according to the US Centers for Disease Control and Prevention (CDC). Three out of four sero-postive persons have also shown to be viremic, and thus this corresponds to an estimated 2.7 million people with active infections of HCV worldwide. Infection due to HCV accounts for 20% of all acute hepatitis cases, as well as an estimated 30,000 new acute infections; it has been credited for some 8,000 to 10,000 deaths per year in the United States [7] 

1.8 Prevalence of HCV in Pakistan:

Hepatitis C virus is one of the most common blood-borne viruses and is associated with significant morbidity and mortality. It affects 170 million people worldwide and 2.4%-6.5% people in Pakistan. Like the other countries of South-East Asia, the number of patients afflicted with hepatitis C in Pakistan is increasing day by day. Unfortunately, most studies assessing the burden of HCV are carried out in particular settings or among blood donors. Hence, their results cannot be generalized, the actual magnitude remains mostly concealed and there is variability in the reported prevalence throughout the country. A majority of the reported prevalence estimates for HCV range between 2.4%-6.5% among adults and 0.44-1.6% among children. Furthermore, among adults HCV seroprevalence was 4.57% in patients asymptomatic for liver disease in Buner, NWFP, 5.31% in Islamabad, 15.9% in Lahore and 23.8% in Gujranwala. Highest HCV seropositivity from Pakistan is 20.6% in Faisalabad and 44% from Darsano Channo, Punjab. Hence, the reported HCV prevalence in Pakistan is much higher when compared to the other countries of the region like India (0.9%), Indonesia (2.1%) and China (3.2%). Blood donors are generally considered a healthier segment of the community, and hence considered representative of the general population. The reported prevalence of HCV among blood donors from Pakistan ranged between 1.18% to 7.5%, whereas regional HCV prevalence among blood donors was reported as 4.1% in Peshawar,19 7.5% from the interior of Sindh, 1.18-3.6% from Karachi, 68% from Lahore and 4% from Rawalpindi. This difference in intra-regional HCV seropositivity amongst the donor population may have been due to different characteristics of the prevalent blood donors; 20% were paid blood donors and 2.4%-3.6% were replacement blood donors. Nevertheless, these studies have limitations; most of them included men, young or middle aged people, and therefore may not be representative of general population. Even then, these reported prevalence estimates among blood donors are much higher than those reported from India (1.85%-2.5%) and developed countries.

Healthcare professionals in Pakistan are also at higher risk of acquiring HCV then the general population. Even though the prevalence of HCV in Pakistan is 4-5.6% in healthcare professionals, the figure is 20-fold higher than global prevalence. Furthermore, among intravenous drug users who are at higher risk of acquiring blood-borne viruses, 60-94.3% is affected with hepatitis C, and this figure is very high when compared to the developed world. [8] 

1.9 Genotypes:

The Term genotype refers to different genetic variations or strains of hepatitis C. The variance in genetic differences is approximately 1/3 between the different genotypes. There are six major groups or genotypes numbered 1 to 6 although some experts believe that there may be as many as 11.

HCV genotypes and subtypes are distributed differently in different parts of the world, and certain genotypes predominate in certain areas. Genotypes 1-3 are widely distributed throughout the world. Subtype 1a is prevalent in North and South America, Europe, and Australia. Subtype 1b is more common in North America and Europe, and is also found in parts of Asia. Genotype 2 is present in most developed countries, but is less common than genotype 1. Some studies suggest that different genotypes and subtypes of HCV may be associated with different transmission routes. For instance, subtype 3a appears to be prevalent among injection drug users and it is believed that it was introduced into North America and the United Kingdom with the widespread use of heroin in the 1960s. [9] 

HCV Genotype Distribution

HCV Genotype Distribution

1, 2, 3

Worldwide

4

Middle East, Africa

5

South Africa

6

Southeast Asia

Table No. 02 Worldwide Distribution of HCV

ViralCancer15

Figure No. 02

The predominant HCV genotype in Pakistan is type 3a followed by 3b and 1a. The frequency of genotype 1 was observed to be increasing in this country without any increase in the frequency of genotype 3 that may be very dangerous in coming 15-20 years. Regional difference in genotypes was observed only in Baluchistan province of Pakistan. More than 70% of the cases in Pakistan are hospitals acquired. [10] 

HCV 3 was the leading genotype in Pakistan. Other major types were genotype 1 and mixed genotype. A high prevalence of variant of genotype 3 alone to be over 79.43% in Pakistan overall- alone or in combination with another genotype. Genotype 3 requires shorter duration of treatment as compared with genotype 1, with its associated reduced cost and side effects. The predominance of HCV genotype 3 in our population confirmed the predominance of HCV genotype 3 in the surrounding countries including India, Iran, Bangladesh and China. Genotype 1 is the second highest genotype in the country. Baluchistan shares a long border with Iran in the west where genotypes 1a and 3a are most prevalent. High prevalence of genotype 3a and 1b has been reported from China. It is quite possible that genotype 1 may have entered into Pakistan from these countries through local persons who cross borders for job and trade. [11] 

Meanwhile the distribution of blood group in Pakistan is that the blood B positive (RH) is higher

Then the other blood groups and the Hcv positive patient are also higher than other blood group.

2).Distribution of blood group worldwide:

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AIMS & OBJECTIVES:

Study on the distribution of hepatitis C (Hcv) & genotyping their relation to blood groups and rhesus (RH) factor .

CHAPTER 02

REVIEW OF LITERATURE

Part A: for HCV genotyping

Part B: Blood group and associate disease

Part (A) For Hcv genotyping

In 2012, Asher Zaman and his colleagues stated the Genotype3 was the most prevalent in 73 samples (65.17%) followed by genotype 1 in 24 (21.42%) and genotype 2 in 13 (11.60%) samples. Genotype3 had significantly high prevalence (P=0.000 2). The results showed that 48 (42.85%) samples were infected with HCV 3a; 25 (22.32%) with 3b; 14 (12.50%) with 1a; 10 (8.92%) with 1b; 11 (9.82%) with 2a; 2 (1.78%) with 2b; and 2 were untypable. The distribution ofHCVgenotypes in Mardan, Charsadda, Peshawar, Sawabi and Nowshehra districts was different. Use of unsterile equipment for medication, barbers and previous history of hospitalization were the main risk factors for HCV transmission [12] 

Anita Chakarvarti in 2011 stated the 300 chronic liver diseases patients screened for the presence of anti-HCV antibodies, 145 were positive. These HCV antibody positive patients were tested for the presence of HCV RNA and 71 patients were found to be HCV RNA positive. All HCV RNA positive samples were subjected to genotype determination. The analysis revealed the presence of genotypes 1, 2 and 3 using RFLP and type specific PCR followed by direct sequencing. The genotype 3 was observed in 45 (63.38%) patients. Of these, 31 showed infection with subtype 3a (68.8%), 12 had subtype 3b (26.6%) and 2 patients showed unique subtype 3i and 3f (4.44%). Genotype 1 was seen in 22 (30.98%) patients. Of these 22 cases, 8 had subtype 1a, 10 had subtype 1b and only 5 patients had subtype 1c infection. Genotype 2a was seen in four patients only [13] 

In 2011 Slawomir Chlabicz and his colleagues studied that Among 290 patients with HCV infection who had HCV genotype determined, there were 45 (15.5%) patients infected with genotype 4, 146 (50.3%) with genotype 1 and 99 (34.1%) with HCV 3. The majority of patients with genotype 4 had similar characteristics to those infected with other genotypes and were males (69%) aged between 31 and 50 years of age (66.7%) poorly educated (basic education or lower secondary 55.6%)). The mean age of patients with HCV 4 and those infected with other than HCV 4 genotype was not significantly different (34 and 36.9 years, respectively). Overall 60% of HCV 4 infections occurred in patients with prior intravenous drug use while 37.6% of infections with other genotypes were drug-related. In 119 patients whose source of infection was other than drug use, there were 16 (10.5%) HCV 4 cases. The majority (51%) of HCV 4 individuals were also HIV-positive (28.9% in case of other genotypes). (Among 94 patients with HCV/HIV co-infection, HCV 4 was present in 23 (24.5%) individuals. Thirteen of HCV 4-positive patients who were HIV-negative and never received treatment for HCV (9 men, 4 women, mean ± SD age of 41.6 ± 11.9 years) received combined antiviral treatment (Peg-IFN α-2a 120-180 µg/week, Peg-IFN α-2b 100-150 µg/week or natural IFN α 9 MU/week with ribavirin (800-1200 mg/day based on body weight). HCV RNA was assessed at baseline, and after 3, 12 and 18 months of start of the treatment. In three (23%) patients the treatment was stopped after three months due to lack of response (defined as at least 2 log10 reduction in HCV RNA at week 12). End-of-treatment viral response (defined as negative for HCV RNA at the end of 48 week therapy) was achieved in nine (69%) patients and sustained viral response (defined as negative for HCV RNA six months after end of therapy) in six (46%) patients. [14] 

Aamir et al, 2009 in a study stated that Hepatitis C Virus (HCV) is thought to be one of the major causative agent of viral hepatitis. A large number of HCV infected patients develop chronic hepatitis which often results in liver cirrhosis and even progress to hepatoma The HCV genotype is the strongest predictive parameter for sustained virological response (SVR). Patients with different HCV genotypes respond differently to antiviral therapy. Firm evidence has been established that patients with genotypes 2 and 3 are more likely to achieve SVR to combination therapy than Genotype 1 patient. [15] 

Wasim Jafri, Amna Subhan, 2008 stated that Hepatitis C virus is one of the most common blood-borne viruses and is associated with significant morbidity and mortality. It affects 170 million people worldwide and 2.4%-6.5% people in Pakistan. Therapeutic injections by contaminated, re-used syringes, transfusion of unsafe blood and re-use of razors are major factors responsible for the spread of hepatitis C in the general population. Genotype 3 is the most common genotype in Pakistan and is most responsive to interferon and ribavirin combination therapy. HCV is the leading cause of chronic liver disease and hepatocellular carcinoma in Pakistan. Appropriate steps need to be taken in the country to control factors responsible for the spread of hepatitis C [16] 

Muhammad Idrees and Sheikh Riazuddin, 2008 stated the most common HCV genotype in Pakistan is type 3a. Regional difference in genotypes was observed only in Balochistan province of Pakistan. More than 70% of the cases were acquired in hospitals through reuse of needles/syringes and major/minor surgery that is very common in this country. [17] 

(Tomoko Takeuchi, 2005) The establishment of a real-time detection system enables more accurate diagnosis of infection and monitoring of viral load in interferon-treated patients via quantification of viral genome. [18] 

(Patrizia Farci, 1991) Their data indicate that HCV RNA is a crucial marker for establishing a diagnosis of primary HCV infection early in the course of infection and discriminating between past and active HCV infection in patients with persistent or fluctuating antibody patterns. In addition, testing for HCV RNA has prognostic value; sustained clearance of HCV RNA correlates with resolution of the disease, whereas persistence of detectable HCV RNA predicts progression to a chronic form of hepatitis.

The clinical detection of HCV infection has evolved rapidly over the last 10 years, and today many of the latest diagnostic tools, including quantification of HCV-RNA levels, may prove invaluable in guiding therapeutic intervention and/or therapy customized for individual patients. Quantification of HCV-RNA levels, determination of HCV genotypes, and measurement of HCV quasispecies have been employed to characterize the natural history of HCV disease, and have opened up new areas for active investigation [19] .

104 patients (58%) had subtype 1a; 38 (21%) had subtype 1b; 4 (2%) had subtype 2a; 23 (13%) had subtype 2b; 8 (5%) had subtype 3a; and 2 (1%) had subtype 4a. Examination of the known risk factors for acquiring HCV showed no association between genotype and mode of acquisition (blood transfusion, injection drug use, employment at a health care facility) or histologic findings at presentation (mild active hepatitis, moderately active hepatitis, or cirrhosis). Sixty-eight percent of patients with genotype 1a, 80% of patients with genotype 1b, and 37% of patients with genotype 2a or 2b had severe hepatitis. Thirteen of 46 (28%) patients with genotype 1a and 4 of 15 (26%) patients with genotype 1b had a complete biochemical response after 6 months of interferon therapy. In contrast, 10 of 14 (71%) patients with genotype 2a or 2b had a complete response to interferon therapy. Five of 39 (13%) patients with genotype 1a, 1 of 14 (7%) patients with genotype 1b, and 2 of 11 (18%) patients with genotype 2a or 2b had a sustained biochemical response. [20] 

Fifty-seven (79.2%) of 72 patients with HCC had genotype 1b HCV, whereas 101 (77.1%) of 131 patients without HCC had genotype 1b, indicating that there was no significant difference in the prevalence of genotype 1b HCV between the patients with and without HCC. Furthermore, comparison of patients with HCC with genotype 1b HCV with those with genotype 2a who had a history of blood transfusion did not differ significantly in the number of years from blood transfusion to diagnosis of HCC. Levels of HCV-RNA were not significantly different among patients with liver diseases of various stages.

Part (B) Blood group and associate disease:

1. ABO blood groups have shown some association with various diseases. Although there are small studies in literature about association between ABO blood groups and chronic viral hepatitis, only few studies found relation between fibrosis severity in chronic viral hepatitis C (CVHC) and ABO blood groups. The aim of this survey is to determine the frequency of different blood groups and Rh (Rhesus) factor in chronic viral hepatitis B (CVHB) and C, to find out if such a host factor may play a role in trend of these viral infections. [21] 

A and B antigens are secreted by the cells and are present in the blood circulation. It seems that non secretors are susceptible to a variety of infections. The possible pathogenesis for this susceptibility is that as many organisms that may bind to polysaccharide on cells and soluble blood group antigens may block this binding (Jefferys et al., 2005) [22] 

study about fibrosis severity in chronic viral hepatitis C in association with blood groups showed that group A, B and AB, ABO were associated with more thrombotic events. They concluded that non-O blood groups may have an increasing risk of venous thrombosis (Armelle et al., 2006). In addition non - O blood groups were associated with increase fibrosis [23] 

In one study about ABO blood groups distribution in serum hepatitis (Hepatitis B) a disproportionate excess of blood group O was found in an outbreak of hepatitis B among patients and staff of a Hemodialysis Unit and more severe cases were also mostly of group O (Lewkonia, 1969). [24] 

THE study was performed based on presence of Rh and ABO blood groups which could be used as an index for studying prevalence of infection. The highest prevalence of HIV and HBV infection were found in individuals with blood group O and Rh positive. [25] 

MALIGNANT TUMORS OF THE DIGESTIVE TRACT IN BOSNIA AND HERZEGOVINA (a) THE men became ill from gastric cancer significantly more frequently than women; (b) the frequency of liver carcinoma was three times higher than the global frequency and the frequency neighbouring ethnic groups; and (c) patients with blood group B and patients with RhD (-) exhibited a significantly higher proportion of disease. [26] 

Significantly higher number of RhD positive donors had HCV infection as compared to RhD negative donors (8.25% vs. 3.66%). Conclusion: High frequency of HCV infection in blood donors needs implementation of strict screening policy for donors and public awareness campaigns about preventive measures to reduce the spread of this infection as well as other transfusion transmissible infections. Association of HCV infection with blood group types needs more studies to get more knowledge about this aspect [27] .

ABO blood groups are one set of agglutinogens, which are genetically determined carbohydrate molecules carried on the surface membranes of the red blood cells. ABO blood groups have shown to have some association with various non infectious and infectious diseases. In most people A and B antigens are secreted by the cells and are present in the blood circulation. It seems that non secretors are susceptible to a variety of infections. The possible pathogenesis for this susceptibility is that as many organisms that may bind to polysaccharide on cells and soluble blood group antigens may block this binding (Ahmed et al., 2004). [28] 

Frequency of Hepatitis B surface Antigen (HBsAg) and Hepatitis C virus Antibodies (Anti-HCV) among blood donors of Kolar and their association with blood group types will help us in understanding the pathogenesis of the disease better. [29] 

Chapter - 3

The research methodology

3.1 Introduction

The research bears meaningful and purpose oriented approach. In this way, researchers apply skillful method of science to the art of management. Uncertainty is key element in the game of business. It cannot be reduced by any specific method. Only in this way, the research methodology is used to minimize the probability of making a wrong choice amongst alternative course of action.

3.2 meaning of research

By research, we mean the phenomenon of arranging the attempts to provide the requisite information for making healthy management decisions.

3.3 role of research in important areas

The role of research in the important areas of management has been briefly covered, includes marketing, production, banking, material, Human Resource Development and Government.

3.4 process of research

The process of research involves the following steps.

Definition of problem

Design of research

Accumulation of data

Scrutiny of data

Briefing of outcomes

3.5 research design

The research is a powerful and meaningful activity. This sort of research is categorized as survey research which can only be conducted with the help of already attained material with relation of topics, from a lot of other sources like articles, journals and books written on the concerned research. The questionnaire distributed to various Health Care Facility Providers, Clinic and Hospital by attending the health care facilities personally.

The accumulation of data while bring questionnaires into used and analyzed on the ground of the objective. There is sufficient flexibility in attaining data as unavailability of response is quite low whereas the command and control is possible.

Various graphical and mathematical functions were used to get the complete results and interpreted accordingly. Biased approach is not fruitful for the functioning of research method hence this element must be wiped out.

3.6 the population

A statistical population is defined as "The aggregate or totality of all individual members or objects whether animate or inanimate concrete or abstract of some characteristic of interest".

In our research, all donors came in blood bank for donate blood .

3.7 the sample

The samples were selected on the basis of convenience and judgment. In this research, only Hcv positive donors are selected

3.8 sample size

The method employed in this research was based on interviewing the questionnaire on personal grounds. There is sufficient flexibility in obtaining data as unavailability of response is quite low whereas the command and control is possible.

100 Hcv positive cases is study .

3.9 sampling technique

3.9.1 cross sectional descriptive study

In this study we collect data in a specific time interval.

STUDY SETTING:

Department of Microbiology, IPH Lahore.

3.9.2 formal permission

Owing to be a researcher, working within the parameters of ethics, verbal permission was taken from every sample and also explained the purpose of research and the benefits that could be achieved.

3.9.3 the primary data collection

The primary data was collected by the above cited location. The process employed is given below.

3.10 method of data collection

The method used in this research was personal interview with questionnaire. There is sufficient flexibility in obtaining data as unavailability of response is quite low whereas the command and control is possible.

Time pressure was there overall positive responses were received on the all Health Care Facilities Providers.

3.11 the research instrument

Questionnaire was developed for this study. It was initially PRE-TESTED in order to find out any draw back or discrepancies. Later on a final questionnaire was prepared. The sequence of question was logical in order to achieve the results.

3.12 socio-economic and demographic status of the individual

It was helpful on Two to three accounts for individuals on following;

Age segment

Education

Marital status

Whether the individual was providing health related services in Public and private sector.

Income status for the individual.

Such sort of research was helpful in responding the quarries pertaining to the research.

3.13 secondary data

The researcher had endeavored up to his entire satisfaction to accumulate secondary data from reliable sources such as books, articles and journals. The researcher has chosen the necessary data with respect to desire research in these secondary sources.

3.14 INCLUSION CRITERIA

All patients which have blood group B (RH)positive& positive for HCV PCR.

3.15 EXCLUSION CRITERIA

The sample negative for HCV PCR