@article{a2932f90f85011ddb219000ea68e967b,
title = "Direct evidence of extensive diversity of HIV-1 in Kinshasa by 1960",
abstract = "Human immunodeficiency virus type 1 (HIV-1) sequences that pre-date the recognition of AIDS are critical to defining the time of origin and the timescale of virus evolution. A viral sequence from 1959 (ZR59) is the oldest known HIV-1 infection. Other historically documented sequences, important calibration points to convert evolutionary distance into time, are lacking, however; ZR59 is the only one sampled before 1976. Here we report the amplification and characterization of viral sequences from a Bouin's-fixed paraffin-embedded lymph node biopsy specimen obtained in 1960 from an adult female in L{\'e}opoldville, Belgian Congo (now Kinshasa, Democratic Republic of the Congo (DRC)), and we use them to conduct the first comparative evolutionary genetic study of early pre-AIDS epidemic HIV-1 group M viruses. Phylogenetic analyses position this viral sequence (DRC60) closest to the ancestral node of subtype A (excluding A2). Relaxed molecular clock analyses incorporating DRC60 and ZR59 date the most recent common ancestor of the M group to near the beginning of the twentieth century. The sizeable genetic distance between DRC60 and ZR59 directly demonstrates that diversification of HIV-1 in west-central Africa occurred long before the recognized AIDS pandemic. The recovery of viral gene sequences from decades-old paraffin-embedded tissues opens the door to a detailed palaeovirological investigation of the evolutionary history of HIV-1 that is not accessible by other methods.",
author = "Michael Worobey and Marlea Gemmel and Teuwen, {Dirk E} and Tamara Haselkorn and Kevin Kunstman and Michael Bunce and Jean-Jacques Muyembe and Kabongo, {Jean-Marie M} and Kalengayi, {Rapha{\"e}l M} and {Van Marck}, Eric and Gilbert, {M Thomas P} and Wolinsky, {Steven M}",
note = "Keywords: Adult; Canada; Democratic Republic of the Congo; Evolution, Molecular; Female; Genetic Variation; HIV Infections; HIV-1; History, 20th Century; Humans; Male; Microtomy; Molecular Sequence Data; Paraffin Embedding; Phylogeny; Reverse Transcriptase Polymerase Chain Reaction; Sequence Analysis, DNA",
year = "2008",
doi = "10.1038/nature07390",
language = "English",
volume = "455",
pages = "661--4",
journal = "Nature",
issn = "0028-0836",
publisher = "nature publishing group",
number = "7213",
}