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. 2021 Feb;73(2):861-864.
doi: 10.1002/hep.31453. Epub 2021 Jan 27.

Preliminary Evidence for Hepatitis Delta Virus Exposure in Patients Who Are Apparently Not Infected With Hepatitis B Virus

Isabelle Chemin  1 , Flor H Pujol  2 , Caroline Scholtès  1   3 , Carmen L Loureiro  2 , Fouzia Amirache  4 , Massimo Levrero  1   5 , Fabien Zoulim  1   5 , Jimena Pérez-Vargas  4 , François-Loïc Cosset  4
Affiliations

Preliminary Evidence for Hepatitis Delta Virus Exposure in Patients Who Are Apparently Not Infected With Hepatitis B Virus

Isabelle Chemin et al. Hepatology. 2021 Feb.
No abstract available

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Figures

FIG. 1
FIG. 1
Detection of HDV exposure among 160 Venezuelan patients who were HCV infected. 1HBsAg (SURASE B‐96; General Biologicals Corporation and Architect, Abbott) and anti‐HBc (ANTICORASE B‐96; General Biologicals Corporation and Architect, Abbott) serological markers were tested by commercial immunoassays, and two sera were found negative by all the tests. 2No HBV DNA (lower limit of detection [LLOD] 50 copies/mL) was detected by qPCR (IQ SYBR Green, Bio‐Rad, using primers located in the HBV S gene (sense [Ss: 5′‐GTG TCT GCG GCG TTT TAT CA‐3′, nts 379‐398 bp] and antisense [Sas: 5′‐GAC AAA CGG GCA ACA TAC CTT‐3′, nts 456‐476 bp]) nor by ddPCR (LLOD 10 copies/mL) in either sample. ddPCR was performed on a QX100 Droplet Digital PCR (Bio‐Rad) using the Taqman Pathogen Detection assay ID Pa03453406_s1 from Thermo Fisher Scientific (HBV detection, primer mix for amplification of P, S/P, and X genes). 3OD/cutoff average ratio of anti‐HDV antibody assays obtained by enzyme‐linked immunosorbent assay (ETI‐AB‐DELTAK‐2, DiaSorin). Negative control OD ≥ 0.6; positive control OD ≤ 0.008; cutoff OD = 0.4. OD/cutoff average ratio of negative samples = 2.339. 4AU obtained by anti‐HDV antibody testing with LIAISON murex XL Anti‐HDV, DiaSorin (Ref 311260, indirect enzyme immunoassay). Negative control < 0.02 AU/mL; positive control > 13 AU/mL; cutoff = 1.0 AU/mL. 5HDV RNA was detected by nested PCR using the following primers: 5413: 5′‐GCC CAG GTC GGA CCG CGA GGA GGT‐3′ (nts 858‐881), 8276: 5′‐ACA AGG AGA GGC AGG ATC ACC GAC‐3′ (nts 1312‐1289), 5414: 5′‐GAG ATG CCA TGC CGA CCC GAA GAG‐3′ (nts 883‐906), and 5415: 5′‐GAA GGA AGG CCC TCG AGA ACA AGA‐3′ (nts 1288‐1265). Abbreviations: AU, arbitrary unit; G, HCV genotype; OD, optical density; nts, nucleotides.
FIG. 2
FIG. 2
(A) Phylogenetic analysis of Venezuelan HDV isolate by Maximum Likelihood method (360 nt). The evolutionary history was inferred by using the Maximum Likelihood method based on the General Time Reversible model (1,000 bootstrap replicas). A discrete gamma distribution was used to model evolutionary rate differences among sites (3 categories +G, parameter = 1.3600). Sequences are shown by their accession number and country of origin. Genotypes are shown in different colors. The Venezuelan HDV isolate retrieved in this study (C3712, accession number MT274327) is shown in bold. Sequences from HDV isolates from patients whose RNA was simultaneously extracted and DNA amplified along with the Venezuelan isolate are included and referred to HDV3, HDV5, and HDV7 sequences. The molecular clone of HDV (GenBank number M21012) used as positive control of PCR is included (C+). (B) Amino acid alignment of the C‐terminal region of the HDV antigen. The last amino acids after the amber stop codon (*) are those present only in the HDV large antigen. Sequences from HDV genotype 1 and from the most divergent genotype 3 are shown. Sequence alignment and phylogenetic analysis were conducted using the Molecular Evolutionary Genetics Analysis version 7 software.
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References

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