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Preliminary Study of Molecular Epidemiology on Hepatitis B among Certain Regions in Anhui Province

Author: WangJuan
Tutor: YeDongQing;LiZuoQing
School: Anhui Medical University,
Course: Epidemiology and Biostatistics,
Keywords: Hepatitis B virus Seroepidemiology Survey Integrin alpha V polymorphisms genetic susceptibility Genotypes
CLC: R512.62
Type: Master's thesis
Year: 2011
Downloads: 32
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BackgroundHepatitis B is an infectious disease caused by hepatitis B virus (HBV), characterized by various inflammatory liver diseases and multiple organs damage. It’s prevalent all over the world, and trends to develop chronic hepatitis, cirrhosis and hepatocellular carcinoma (HCC), and threats heath. According to a report from World Health Organization (WHO), 2 billion persons have been infected with hepatitis B virus, about 1 million persons died from cirrhosis, HCC, etc. Consequently, HBV infection is a serious public health problem that seriously affects health.As a matter of fact, prevalance of HBV infecion differed greatly in distinct areas. According to HBsAg carrying rates in humans,the world is divided into three prevalent areas from low carrying rate to high carrying rate, China belongs to the high prevalent area, and different areas in the country also has corresponding prevalence. Once an investigation in the Anhui province showed that it’s prevalent in the Wanjiang territory with rate of 9.8%. Maybe it’s owing to the backward foundation, shortage of knowledge and resource,relatively bad quality of vaccination,as a result,the current prevalance might be severe in the province. Consequently,we will perform an epidemiological investigation in order to comprehend the prevalence of hepatitis B and the state of the art in the rural region in this province.As is known, individuals have different results after HBV infection, besides the effect of the virus,genetic predisposition might be the prominent factor. Integrins are transmembrane glycoprotein receptor families, located on the cell membrane surface, play important roles in the physiological and pathological processes. Several researches suggest that expression of integrins and ECM protein in the liver are related to HBV infection and HCC progression. While in the impaired liver, activated stellate cells produced upregulated integrins. IntegrinαⅤ( ITGAV) belongs to the integrin family, a study on hepatitis B patients in Korea indicates that ITGAV gene polymorphism might be associated with the susceptibility of HBV infection and HCC, which has not been demonstrated in Chinese. Consequently, study on the chronic mechanism of HBV infection in a Chinses Han pupulation, and from the individual perspective of genetic susceptibility to approach the reason of distinct result of HBV infection might have theoretical and pratical significance. In addition, more and more evidences have implied that HBV genotypes are associated with the progression of a patient’s condition, disease’s mode of transmission and clinical turnover. Therefore, we will also detect the genotypes of the patients so as to understand the distribution of the HBV genotype and dissect the relationship between genotype and disease activity as well as progression of the disease. Thus, two parts named current hepatitis B prevalance and genetic susceptibility studies form the whole study.PartⅠA Prevalence and Knowledge Survey on Hepatitis B among A Population in Rural Areas in A City of Anhui ProvinceObjective To investigate the epidemic status of hepatitis B and the status of cognition about hepatitis B in certain rural areas in Anhui province.Methods A cluster sampling method was conducted to perform questionnaire on 2038 persons who were selected randomly from rural population of Hefei and 3ml of the vein blood was drawn from all the individuals engaged in the study. HBsAg, anti-HBs of the samples were tested with ELISA.Results Among 2038 people, the prevalence of HBsAg, anti-HBs infection were 6.44% and 46.85% respectively. The prevalence of HBsAg was not significantly different between males and females (P>0.05). The prevalence rate of HBsAg was 9.06% in rural population, which was higher than that of other groups, the rate was the lowest (2.70%) in students group. The prevalence rate of HBsAg among people who were born after 1992 was significantly lower than that of people born before 1992 (P<0.05).The vaccination rate was 38.87%. The vaccination rate was the highest in student group (85.41%) and the lowest in rural population (12.19%).The average score of rural population was the lowest. The three accesses to Hepatitis B knowledge were television , radio and print media .Conclusions In the study population of the district, the prevalence rate of HBsAg is 6.44%, which is lower than that of nation-wide. As the prevalence rates of anti-HBs and the knowledge on hepatitis B among rural population is relatively low, it is essential to strength health education and widespread immunization with hepatitis B vaccine in this population. Objective 1. To investigate the association of ITGAV(rs1839123) polymorphisms with susceptibility to HBV infection in a Chinese Han population. 2. To investigate the distribution of hepatitis B virus (HBV) genotypes in hospital patients with HBsAg positive in Jieshou city, Anhui province of China, and the relationship between the genotype and clinical patterns and progression of the disease.Methods 1. 314 hepatitis B patients and 476 healthy volunteers matched by sex and age were recruited from Infectious Department of Anhui Provincial Hospital and Jieshou People’s hospital. Diagnosis of all patients with positive for hepatitis B surface antigen (HBsAg) was made on the basis of ELISA. Patients with other types of liver diseases, such as alcoholic liver disease, autoimmune liver disease and metabolic liver diseases were excluded. We also excluded subjects who were positive for anti-HCV. Every respondent in our study was given informed consent. The study protocol was approved by the Department of Epidemiology and Biostatistics at Anhui Medical University. We also excluded subjects who were positive for anti-HCV. Design a survey to collect clinical and laboratory and epidemiologic data of all respondents. Forearm venous blood samples from each subject were collected in 5 ml vacuum tube containing EDTA and citrate, the plasma were excluded from the cell pellet by pipetting after being centrifuged at 2,000 revolutions for 10 min. Genomic DNA was extracted from the cell pellet in whole blood (QIAamp blood kit; Qiagen) and stored at -80℃before test. The polymorphisms of the SNP (rs1839123) were genotyped by RT-PCR (corporation ABI, 7300) with Taqman fluorescent probe. Assess the level of differences in distributions for allelic and genotype frequencies between those with and without chronic hepatitis B. The observed frequencies of the genotypes were compared with the frequencies under Hardy-Weinberg equilibrium byχ2 tests. All statistical analyses were done by SPSS 10.01. All P values were calculated based on 2-sided tests, with statistical significance defined as P value≤0.05.2. The HBV genotypes were tested by polymerase chain reaction (PCR) and fluorescence probe technique,as the serum marker of HBV tested by CMIA and the HBV DNA level by real-time fluorescence quantitative detection.Results 1. (1)HWE test No deviations from HWE were observed in neither the patients with chronic hepatitis B or the controls in each polymorphism.(χ2=0.099, P=0.754;χ2=0.464,P=0.496) (2)Association analysis of ITGAV(rs1839123) and HBV infection Analysis of genotypes of ITGAV (rs1839123) revealed no significant deviation from Hardy–Weinberg equilibrium in any group. The frequency of A and G allele was 17.68% and 82.32% in patients with HBV infection. The genotype frequencies of the SNP in patients were listed as follows: GG, GA and AA were 67.52%, 29.62%, and 2.86%, respectively. Frequency analysis showed that there’s no association between ITGAV (rs1839123) polymorphisms and HBV infection (P>0.05).2. Of the 48 patients, 1 (2.08%) was genotype B, 35 (72.92%) were genotype C, 12 (25.00%) were failed to detect the genotypes. Genotype B,C combination and D was undiscovered. No significant difference was found in sex, age between HBV genotype B and C (P>0.05). There was no significant difference between B and C genotype distribution among patients with HBV carriers, chronic hepatitis B, liver cirrhosis and hepatocellular carcinoma (P>0.05). The positive rate of HBeAg was 42.86% in genotype C group. A significant difference was found in the level of HBV-DNA positive among the genotypes (P<0.05).Conclusions1.No deviations from HWE were observed in either the patients with HBV infection or the controls in polymorphism for SNP rs1839123. No significant differences existed between the patients with HBV infection and the controls, which suggests that the polymorphisms of ITGAV(rs1839123) might not contribute to HBV infection susceptibility in the Chinese population.2. The predominant HBV genotype in the city was confirmed to be genotype C. As there was significant difference in the HBV-DNA level among the genotypes, genotype C may be associated with the progression of liver diseases.

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CLC: > Medicine, health > Internal Medicine > Infectious disease > Viral infections > Viral Hepatitis > Hepatitis B
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