Abstract
Methods: To avoid contamination, viral RNA was extracted in different places (Lyon versus Bobigny and Bangui) for ancient and recent samples, respectively. Manual or automated (Siemens Process Versant kPCR) extractions led to available nucleic acids extracts. Viral RNA presence was based on the RT-PCR of a 400 bp HDV genome fragment (R0 region). Four to six clones were characterized for positive samples. Furthermore we aimed to amplify and clone the full length of HDV coding sequences from R0-positive samples. HD-positive contemporary samples were obtained from a wide serological survey among school and medical students (n = 1335). Results: FH-associated HDV sequences were obtained from 6 patients and indicated that different African HDV1 strains were responsible for the FH outbreak. In one case quasispecies analysis indicated a punctual deletion in the HD coding gene that could lead to a frame shift in the 3′terminal region of the large delta protein gene, changing the carboxy terminal end of the Large HD protein. HBV genotype E (n =8) and A (n =1) corresponded to the circulating HBV strains during this outbreak. Conclusion: HDV viral RNA could be retrieved from ancient serum samples kept at −20°C for 25 years confirming the strengthness of the viral pseudo double stranded RNA. FH delta in Bangui was linked to different HDV strains. However, all corresponded to HDV genotype 1. HBV Genotype E was the main associated HBV genotype already present in RCA in the mid eighties.
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