Survey Of HDV Infection And Molecular Characterization Of HDV,HBV And HIV-1 Among Chronic Hepatitis B Patients In China

BackgroundHepatitis delta virus?HDV?is defective virus,coated with the hepatitis B surface antigen?HBsAg?,as a satellite of HBV.It requires HBsAg for the assembly of virion and establishment of HDV infection.It was shown that most patients with HDV/HBV co-infection take more sever liver disease,show more rapid progression to cirrhosis and hepatic decompensation,and increased death.The prevalence of HBV could be inhibited by the successful HBV vaccination programs,which could furthermore reduce HDV prevalence.However,the recrudescence of HDV infection may occur among people immigrating from endemic areas and/or with high-risk population such as Injection drug users?IDUs?and HIV infection.In China,over 90 million individuals were living with chronic HBV infection,with the prevalence rate of 6%.There is a huge population with HIV infection in China,and the prevalence rate of HIV was high in special province.In Nov.2017,there were 750 thousand HIV-1 infected persons reported.Of whom,about 6.3%-10.9%was infected with HBV.Because HBV and HIV share common routes of transmission,HDV transmission can also spread with HBV among HIV infected individuals and make more sever liver disease.In China,HDV infection could be prevalent due to the high HBV epidemic in the community,but the data is limited.Our study is aiming to exam the HDV infections among chronic hepatitis B patients with or without HIV infection in seven provinces and analysis the risk factors associated with HDV infection in China.It would provide the available data to carry out the valid measure to control the prevalence of HDV.MethodsA two-targeting detection for HDV RNA was developed based the Taqman probes and Real-Time PCR.It was evaluated by the First WHO International Standard for Hepatitis D Virus RNA.A total of 3000 samples from chronic hepatitis B patients with or without HIV-1 infection were collected from seven research provinces of Beijing,Inner Mongolia autonomous region,Xinjiang Uygur autonomous region,Guangxi Zhuang autonomous region,Yunnan province,Sichuan province and Guangdong province.The number of HIV/HBV co-infection individuals was 759.We also collected the clinical information and demographic information.All of the specimens were detected for HDV screening.Using the results,the clinical information and demographic information,the potential high risk factors for HDV infection were evaluated with Chi-square test and Logistic regression analysis.The full genome of HDV was amplified and sequenced from the specimens with HDV RNA positive.The HIV pol gene and HBV S gene were amplified and sequenced to study the pattern of HDV/HBV or HDV/HBV/HIV co-infection and the molecular characterization of virus.ResultsDevelopment of Detection for HDV RNAWe developed a two-targeting detection for HDV RNA with the lower limit of detection was 575 IU/ml.Combining with the detection of HDV antibody,it could be applied in the clinical detection of HDV infection in chronic HBV infected patients in China.The Prevalence of HDV among Chronic Hepatitis B Patients with or without HIV-1 Infection in ChinaIn this study,HDV infection was existed at all of the seven research provinces,with the HDV positive rate was 2.63%.Logistic regression analysis was performed to identify the potential high risk factors for HDV infection,and the region and HIV infection were found.Compared with HIV negative persons,HDV positive rate was significant higher in HIV/HBV co-infected persons?aOR=9.61,95%CI:5.06-18.27?.Compared with Beijing,HDV positive rate was significant higher at the research site of Sichuan?aOR=11.10,95%CI:3.73-33.08?.The HDV positive rate was varied at the different research sites.The HDV positive rate was 13.88%in the research site of Sichuan,which was highest than others.The HDV positive rate was 5.32%in Guangxi Zhuang autonomous region,2.45%in Guangdong province,0.88%in Xinjiang Uygur autonomous region,0.50%in Inner Mongolia autonomous region,0.48%In Beijing,and 0.45%in Yunnan province.It was interesting that,the percent of person with evaluated ASL,ALT,total bilirubin and HBeAb positive in HDV infected people was significant higher than that in HDV negative people.So the liver damage could be induced by HDV,and HDV infection might show negative effects on HBV replication.At the research site of Sichuan,Logistic regression analysis was performed to identified the associated risk factors with HDV infection.In addition to HIV infection,the gender was another associated risk factor with HDV infection.HDV positive rate was higher in male than female.At the research site of Guangxi Zhuang autonomous region,the HDV positive rate was significant higher among IDUs than that among non-IDUs?aOR=31.83,95%CI:4.71-215.09?.In the HIV/HBV co-infected population,the research site at Sichuan was also the high risk factor of HDV infection?aOR=20.67,95%CI:2.56-166.70?,and showed the highest HDV positive rate than others.In chronic hepatitis B patients without HIV-1 infection,the research site of Guangxi Zhuang autonomous region was the high risk factor of HDV infection?aOR=16.74,95%CI:2.50-112.01?,and showed the highest HDV positive rate.The Molecular Characterization of HDV,HBV and HIV-1 among Chronic Hepatitis B Patients with or without HIV-1 Infection in ChinaTwenty HDV full genome sequences were amplified from the HDV RNA positive samples.The obtained sequences were undergone phylogenetic tree analysis.There were only two HDV genotype found,including HDV-1 and HDV-2a.The predominant circulating HDV genotype was HDV-2a in Sichuan.In this study,236 sequences of HIV-1 pol gene were obtained from the HIV/HBV co-infected persons to analysis the molecular characterization of HIV and the association between HIV and HDV.The genotyping was performed by phylogenetic and recombinant patterns analyses.The mainly circulating HIV subtypes were CRF07_BC and CRF01_AE,followed by Subtype B',CRF08_BC,CRF67₀1B,CRF57_BC,CRF65_cpx,CRF68₀1B.Four unique recombinant forms were identified in Beijing and Sichuan.Some difference in HIV subtype distribution were shown in Beijing,Sichuan province and Guangdong province.At the research site of Sichuan,CRF07_BC account the highest proportion,up to 92.5%,followed by CRFO1_AE?3.75%?.At the research site of Beijing,the circulating HIV subtypes were CRF01_AE,CRF07 BC and subtype B',which account for 51.61%,31.18%,and 10.75%,respectively.In addition to CRF07_BC?46%?and CRF01_AE?22%?,CRF55₀1B was also circulating at research site in Guangdong province,accounting for 16%.Cross tabs with Chi-square of independence analyses showed that the distribution of HIV subtypes between HDV positive and HDV negative was similar.In this study,790 sequences of HBV S gene were obtained from chronic hepatitis B patients and analyzed the molecular characterization of HBV and the association between HBV and HDV.The phylogenetic trees were constructed to identified the HBV genotype.The mainly circulating HBV genotype were HBV-C?52.03%?,HBV-B?28.86%?and HBV-D?18.10%?.Two HBV-I and six recombinant forms was obtained from the research sites of Sichuan and Inner Mongolia autonomous region.It showed that the mainly circulating HBV genotypes were significantly different among the different regions.The mainly circulating HBV genotype was HBV-C at the research sites of Beijing,Yunnan province and Inner Mongolia autonomous region.While it was HBV-B at the research sites of Guangdong province and Guangxi Zhuang autonomous region.HBV-B and HBV-C showed the similar proportion at the research site of Sichuan.At the research sites of Xinjiang Uygur autonomous region,HBV-D was the mainly circulating genotypes,which accounted for about 84.11%.The pattern of HDV/HBV co-infection was HDV with HBV genotype B or C,and the pattern of HDV/HIV co-infection was HDV with HIV-1 CRF07 BC.However,the genotype distribution of HBV or HIV between HDV positive and HDV negative population was similar,they were all the mainly circulating genotype in the local.ConclusionsA two-targeting detection for HDV RNA was development,with the lower limit of 575 IU/ml,which could be applied to the clinical analysis.It was the first time to investigate the HDV prevalence among chronic hepatitis B patients with or without HIV-1 infection collected from North,West,Central,South and Southwest of China in recent years.By the study,HDV prevalence was low in China,however,it was significantly high among the HIV-infected individuals in Sichuan?Liangshan Yi Autonomous Prefecture?,and IDUs in Guangxi Zhuang autonomous region.HDV infection could induce liver damage and worsen disease.It is urgent to carry out the available HBV vaccinating programs in Liangshan Yi Autonomous Prefecture,especially for the HIV/HBV co-infections and IDUs,to control the HBV and HDV prevalence.This study provides scientific data to support the prevention and control of HDV epidemic in mainland China.