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Hepatitis D virus infection in Thailand: HDV genotyping by RT-PCR, RFLP and direct sequencing
A Theamboonlers1, T Hansurabhanon, V Verachai
1Dept. of Pediatrics, Faculty of Medicine, Chulalongkorn University & Hospital, Bangkok, Thailand.
Insights
Hepatitis D virus (HDV) infection remains limited among intravenous drug users in Thailand, with detected cases predominantly being genotype I. This suggests a similar pattern to Western HDV genotype I strains.
Area of Science:
- Virology
- Hepatology
Background:
- Hepatitis D virus (HDV) is an RNA virus requiring hepatitis B virus (HBV) surface antigen for infectivity.
- HDV has three classified genotypes, with prevalence in Thailand noted among intravenous drug users (IVDU).
Purpose of the Study:
- To determine HDV genotypes prevalent in Thai IVDU population.
- To analyze the genetic characteristics of HDV in this specific demographic.
Main Methods:
- Screening of 743 IVDU sera for HBV infection.
- Serological analysis for anti-HDV in HBsAg-positive samples.
- Restriction fragment length polymorphism (RFLP) and direct sequencing of amplified HDV genome.
Main Results:
- 7% of IVDU sera were HBsAg-positive; 21.8% of these were positive for anti-HDV.
- HDV-RNA was detected in 66% of anti-HDV positive specimens.
- All analyzed HDV samples were identified as genotype I, confirmed by direct sequencing.
Conclusions:
- HDV infection prevalence is limited within the studied IVDU population in Thailand.
- The observed HDV genotype pattern aligns with Western HDV genotype I strains.
Background:
Hepatitis D virus (HDV) is a degenerate RNA virus or virusoid that requires the surface coat of hepatitis B virus (HBV), i.e. hepatitis B surface antigen (HBsAg), in order to become infectious. Three distinct genotypes of the virus have been classified. In this study, HDV genotypes were determined by restriction fragment length polymorphism (RFLP) and direct sequencing. In Thailand, simultaneous HDV/HBV infections are particularly prevalent among intravenous drug users (IVDU).
Patients And Methods:
A total of 743 IVDU sera were screened for HBV infection. HBsAg-positive samples were subjected to serological analysis for anti-HDV. RFLP analysis using the endonucleases Xho I and Sma I was performed on the PCR amplified HDV genome to establish the prevailing HDV genotypes.
Results:
55 sera (7%) had detectable HBsAg; all 55 were subsequently subjected to serological analysis for anti-HDV, 12 (21.8%) of which were positive. Eight (66%) specimens had detectable HDV-RNA by RT-PCR. All polymorphisms were shown to be genotype I, a finding confirmed by direct sequencing. 36 HBsAg-positive sera obtained from the blood bank to serve as controls were negative for anti-HDV.
Conclusion:
Our data show that HDV infection is still limited among IVDU and that the pattern of polymorphism closely resembles that of the western HDV genotype I.
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