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2.
Nature ; 557(7705): 418-423, 2018 05.
Article in English | MEDLINE | ID: mdl-29743673

ABSTRACT

Hepatitis B virus (HBV) is a major cause of human hepatitis. There is considerable uncertainty about the timescale of its evolution and its association with humans. Here we present 12 full or partial ancient HBV genomes that are between approximately 0.8 and 4.5 thousand years old. The ancient sequences group either within or in a sister relationship with extant human or other ape HBV clades. Generally, the genome properties follow those of modern HBV. The root of the HBV tree is projected to between 8.6 and 20.9 thousand years ago, and we estimate a substitution rate of 8.04 × 10-6-1.51 × 10-5 nucleotide substitutions per site per year. In several cases, the geographical locations of the ancient genotypes do not match present-day distributions. Genotypes that today are typical of Africa and Asia, and a subgenotype from India, are shown to have an early Eurasian presence. The geographical and temporal patterns that we observe in ancient and modern HBV genotypes are compatible with well-documented human migrations during the Bronze and Iron Ages1,2. We provide evidence for the creation of HBV genotype A via recombination, and for a long-term association of modern HBV genotypes with humans, including the discovery of a human genotype that is now extinct. These data expose a complexity of HBV evolution that is not evident when considering modern sequences alone.


Subject(s)
Evolution, Molecular , Hepatitis B virus/genetics , Hepatitis B virus/isolation & purification , Hepatitis B/virology , Phylogeny , Africa , Animals , Asia , Europe , Genotype , Hepatitis B virus/classification , History, Ancient , History, Medieval , Hominidae/virology , Human Migration/history , Humans , Recombination, Genetic
3.
J Virol Methods ; 189(1): 41-6, 2013 Apr.
Article in English | MEDLINE | ID: mdl-23318370

ABSTRACT

In the current study, primers described previously and modified versions of these primers were evaluated for amplification of full-length gag genes from different equine infectious anemia virus (EIAV) strains from several countries, including the USA, Germany and Japan. Each strain was inoculated into a primary horse leukocyte culture, and the full-length gag gene was amplified by reverse transcription polymerase chain reaction. Each amplified gag gene was cloned into a plasmid vector for sequencing, and the detectable copy numbers of target DNA were determined. Use of a mixture of two forward primers and one reverse primer in the polymerase chain reaction enabled the amplification of all EIAV strains used in this study. However, further study is required to confirm these primers as universal for all EIAV strains. The nucleotide sequence of gag is considered highly conserved, as evidenced by the use of gag-encoded capsid proteins as a common antigen for the detection of EIAV in serological tests. However, significant sequence variation in the gag genes of different EIAV strains was found in the current study.


Subject(s)
Equine Infectious Anemia/diagnosis , Genes, gag , Infectious Anemia Virus, Equine/genetics , Reverse Transcriptase Polymerase Chain Reaction/veterinary , Animals , Base Sequence , Cells, Cultured , Equine Infectious Anemia/virology , Gene Amplification , Genetic Variation , Horses , Molecular Sequence Data , Sequence Alignment , Sequence Analysis, DNA
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