title: Forensic application of phylogenetic analyses - Exploration of suspected HIV-1 transmission case.
abstract: Transmission of human immunodeficiency virus (HIV) between individuals may have important legal implications and therefore may come to require forensic investigation based upon phylogenetic analysis. In criminal trials results of phylogenetic analyses have been used as evidence of responsibility for HIV transmission. In Serbia, as in many countries worldwide, exposure and deliberate transmission of HIV are criminalized. We present the results of applying state of the art phylogenetic analyses, based on pol and env genetic sequences, in exploration of suspected HIV transmission among three subjects: a man and two women, with presumed assumption of transmission direction from one woman to a man. Phylogenetic methods included relevant neighbor-joining (NJ), maximum likelihood (ML) and Bayesian methods of phylogenetic trees reconstruction and hypothesis testing, that has been shown to be the most sensitive for the reconstruction of epidemiological links mostly from sexually infected individuals. End-point limiting-dilution PCR (EPLD-PCR) assay, generating the minimum of 10 sequences per genetic region per subject, was performed to assess HIV quasispecies distribution and to explore the direction of HIV transmission between three subjects. Phylogenetic analysis revealed that the viral sequences from the three subjects were more genetically related to each other than to other strains circulating in the same area with the similar epidemiological profile, forming strongly supported transmission chain, which could be in favour of a priori hypothesis of one of the women infecting the man. However, in the EPLD based phylogenetic trees for both pol and env genetic region, viral sequences of one subject (man) were paraphyletic to those of two other subjects (women), implying the direction of transmission opposite to the a priori assumption. The dated tree in our analysis confirmed the clustering pattern of query sequences. Still, in the context of unsampled sequences and inherent limitations of the applied methods, we cannot unambiguously prove that HIV-1 transmission occurred directly between two individuals. Further exploration of the known and suspected transmission cases is needed in order to define methodologies and establish their reliability.
authors.affiliation: Institute of Microbiology and Immunology, University of Belgrade School of Medicine, Belgrade, Serbia.
authors.forename: Marina
authors.initials: M
authors.lastname: Siljic
authors.affiliation: Infectious and Tropical Diseases University Hospital, Clinical Center Serbia, HIV/AIDS Unit, Belgrade, Serbia.
authors.forename: Dubravka
authors.initials: D
authors.lastname: Salemovic
authors.affiliation: Institute of Microbiology and Immunology, University of Belgrade School of Medicine, Belgrade, Serbia.
authors.forename: Valentina
authors.initials: V
authors.lastname: Cirkovic
authors.affiliation: Virology Laboratory, Microbiology Department, Clinical Center Serbia, Belgrade, Serbia.
authors.forename: Ivana
authors.initials: I
authors.lastname: Pesic-Pavlovic
authors.affiliation: Institute of Microbiology and Immunology, University of Belgrade School of Medicine, Belgrade, Serbia; Infectious and Tropical Diseases University Hospital, Clinical Center Serbia, HIV/AIDS Unit, Belgrade, Serbia.
authors.forename: Jovan
authors.initials: J
authors.lastname: Ranin
authors.affiliation: Institute of Microbiology and Immunology, University of Belgrade School of Medicine, Belgrade, Serbia.
authors.forename: Marija
authors.initials: M
authors.lastname: Todorovic
authors.affiliation: Institute of Forensic Medicine, University of Belgrade School of Medicine, Belgrade, Serbia.
authors.forename: Slobodan
authors.initials: S
authors.lastname: Nikolic
authors.affiliation: Institute of Microbiology and Immunology, University of Belgrade School of Medicine, Belgrade, Serbia; Infectious and Tropical Diseases University Hospital, Clinical Center Serbia, HIV/AIDS Unit, Belgrade, Serbia.
authors.forename: Djordje
authors.initials: D
authors.lastname: Jevtovic
authors.affiliation: Institute of Microbiology and Immunology, University of Belgrade School of Medicine, Belgrade, Serbia. Electronic address: mstanojevic@med.bg.ac.rs.
authors.forename: Maja
authors.initials: M
authors.lastname: Stanojevic
mesh_terms: D005260:Female; D015658:HIV Infections / Q000635:transmission* / Q000821:virology; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D016013:Likelihood Functions; D008297:Male; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000235:genetics
chemical_list: D012367:RNA, Viral
keywords: Forensic science; Human immunodeficiency virus; Phylogenetic analyses; Transmission cluster; Transmission direction