Seroepidemiological And Molecular Epidemiological Studies Of Viral Hepatitis B After20Years From The Inroduction Of Hepatitis B Vaccine In Insular Regions Of Southeast China

This study was divided into two parts.Part A:Hepatitis B seroepidemiologic investigation in island regionsObjectiveWe expect to learn about the epidemic situation of HBV among all-aged population in the island regions in Zhejiang Province through this seroepidemiologic investigation. Meanwhile we can learn how the expanded program immunization (EPI) carried out in1992affected the epidemic situation of HBV in this region. Furthermore, we compare our result with epidemic situation in plain region in HBV-DNA, hepatic function, AFP, B ultrasound. We learn the epidemic situation of HBeAg in different regions and the relevance between serum marks, HBV-DNA and ALT. This research has vital theoretical and practical significance in evaluating the effects of prophylaxis and vaccination, adjusting of the HBV immunization strategy in time, guiding the government work in the12th Five-Year Plan in Zhejiang province.MethodsStratified clustered-random sampling in different stages was employed in this study. A total of14997residents all-aged were surveyed and blood tested. HBsAg, antibody of HBsAb, and anti body of HBcAb were detected with ELLSA method. ALT and AFP was tested in all the participants and those who were HBsAg positive. ResultsIn14997residents surveyed, we find1763residents are HBsAg positive, the adjusted positive rate is10.4%. The adjusted HBcAb positive rate is33.1%.The adjusted positive rate of HBsAg, HBsAb, HBcAb of male is13.51%、56.9%、39.3%. The adjusted positive rate of HBsAg, HBsAb, HBcAb of female is10.6%、57.0%.40.2%. The rate of elevated ALT is8.2%(12.0%in male,5.8%in female). The rate of elevated ALT among the group over30years old with HBsAg positive is0.6%. The epidemic situation between different islands has significant diversity.ConclusionAccording to three national Viral hepatitis serum epidemiological investigations in1979,1992and2006separately, Chinese Hepatitis B surface antigen(HbsAg) carrier has a downward trend with years. Comparing to the result of the investigation in2006, the hepatitis B prevalence of Zhejiang province also has a obvious decline. The hepatitis B prevalence of island areas (Zhoushan and Yuhuan), however, is still beyond average of Zhejiang province. For purpose of droping national hepatitis B prevalence to low level, the government should give vaccination to more people, put more money down on the vaccination, especially to people whose age are above20and have negative result of HBV markers, as well as high risk group who haven’t already vaccinated since1992. What’s more, comparing to plain region of China, the government ought to focus more on Hepatitis b transmission condition on active floating population in island regions. Part B:Hepatitis B seroepidemiological survey and stratification analysis Object:The hepatitis B vaccine immunization program has been carried out for20years. To understand the prevalence of serological markers among hepatitis B infections in Zhejiang Province, we made a Hepatitis B seroepidemiological survey and further did stratification analysis among the screened hepatitis B infections in insular areas, controlled them to those in plain areas (further detect hepatitis B three-line quantitative HBV-DNA, liver function, alpha-fetoprotein of AFP, B super). Our aim is to understand the epidemic status and the influencing factors of e antigen between different geographical backgrounds and the correlation between serological markers and HBV-DNA and ALT, which is in order to get the latest data to guide the treatment and prognosis of the groups.Method:(1) Stratified clustered-random sampling in different stages were adopted in this study. First, in Shaoxing County and Yuhuan County, which are on behalf of the two major landforms (plains and islands) in Zhejiang Province, we divided all township/street into three layers of high, medium and low, based on the economic level. Select one township/street in each layer randomly. Secondly, select three villages/communities randomly again in every town/street, based on the economic level. In totally, six townships/streets were taken, in which18villages/communities were selected. A total of8439effective objects were surveyed.(2) The questionnaire survey used the unified questionnaire by the method combined concentrated and households, carried out by the professional investigators face-to-face. Serum samples of each participant were collected at the same time, separated. Then send to test related indicators in cryopreservation.(3) Using fluorescence quantitative PCR, real-timely, dynamicly monitor the HBV-DNA load in gene diagnostic laboratory which is certificated by the Ministry of Health. The machine is the FO-33A PCR instrument offered by Hangzhou Bo Japan Science and Technology Co., Ltd. and reagents were purchased from ACON Biotech (Hangzhou) Co., Ltd.. Meanwhile, serological markers were measured by enzyme-linked immunosorbent assay (ELISA). The enzyme-linked immunosorbent assay kits were produced by Hangzhou aicon Biotechnology Company. Operation was in strict accordance with the kit instructions and the results were read and record by the microplate reader (MULTISKAN MK3). In addition, select the Beckman CX7automatic biochemical analyzer to test ALT, which was purchased from Beijing Leadman Biochemistry Co., Ltd..The results were judged by standards of the kit.(4) Adopt the EpiData3.1software to create a survey database and the data was entered in strict accordance with the requirements of the double-entry. Use the statistical software SPSS13.0recommended by WHO to conduct statistic. Statistical methods using the chi-square test and inspection standards for bilateral a=0.05.P<0.01was considered significant difference.Result:Respondents were HBsAg positive, in which the proportion of40-49year-old crowd was the highest, at33.2%; HBeAg positive rate was18.5%, of which, the population that less than20years old and older than70-year-old crowd, their positive rates were60%and9.2%each. The HBeAg positive rates of men and women were21.9%and14.7%each. Data was significantly different. In Yuhuan and Shaoxing, HBeAg positive rates were21.9%,12.9%. Both have significant difference. HBV-DNA positive rate was91.0%among HBeAg-positive people and was45.4%among HBeAg negative people, which was significantly different. The HBeAg positive rates in population, which HBV-DNA amounts were less than103,103～105,105～107,>107were3.6%,6.0%,41.1%,94.0%. There were a lot of differences of HBeAg positive rates in the populations of the different levels of hepatitis B viral load. Do the Spearman rank correlation analysis with the HBV-DNA and HBeAg and find linear relationship (p <0.01) between the two, but weak correlation (r=0.394<0.4). Do Spearman rank correlation analysis with HBV-DNA and HBeAg among HBeAg-positive patients, find the linear relationship (p<0.01), and it is a strong correlation (r=0.777>0.7). In the population of HBeAg positive, the ALT elevating and the ALT normal people, their HBV-DNA positive rates were95.4%、86.4%respectively, which was significantly different; in the population of HBeAg negative, the ALT elevating and the ALT normal people, their HBV-DNA positive rates were59.6%、41.5%, which was significantly different.Conclusion:Compared with the results of investigation made in1992of Zhejiang province, the age structure of the hepatitis B has largely back goes on to40-49, which prompted the Government that it is important to strengthen the coverage of health examination, which includes the HBV detection, and penetration of health education for the40-49’s rural population, in order to detect and treat disease early. Although studies have shown that there is linear relationship between the HBV-DNA and HBeAg, in the clinical, doctors need to consider HBV-DNA and HBeAg amount integratedly, especially the people which HBeAg negative but the HBV-DNA level high. Combining the HBeAg, HBV-DNA and ALT could make comprehensive evaluation of viral replication and the reactions of the body. In no condition to using the PCR method for the quantitative detection of HBV-DNA, HBeAg and ALT levels could be regarded as leading indicators to judge the severity of HBV activity and infectious.