Molecular investigation of transmission of human immunodeficiency virus type 1 in a criminal case - PubMed (original) (raw)

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Molecular investigation of transmission of human immunodeficiency virus type 1 in a criminal case

R Machuca et al. Clin Diagn Lab Immunol. 2001 Sep.

Abstract

Very few criminal cases involving human immunodeficiency virus type 1 (HIV-1) transmission have been described. We report on an HIV-1 transmission case with a child being infected by an HIV-1-positive man. The objective was to determine through molecular epidemiology and phylogenetic analyses whether HIV-1 from the HIV-1-positive man could be the source of infection in the HIV-1-positive child, as claimed by the authorities. We conducted genetic analysis of three different parts of the HIV-1 genome (gag, pol, and env) by PCR, direct-sequencing, and phylogenetic analyses. We used maximum likelihood, maximum parsimony, and neighbor-joining methods for the phylogenetic analyses to investigate whether the sequences from the man and the child were related. We found that the viral sequences from the man and the child formed separate clusters in all of the phylogenetic analyses compared to the local controls. A unique amino acid deletion was identified in the C2-V3-C3 region of the env gene in the virus from the man and the child. These results were used in the criminal court to elucidate whether the virus from the man was related to the virus from the child. In summary, the results from the phylogenetic analyses, the sequence distances between the virus from the man and the virus from the child, and the identification of the unique molecular fingerprint in the env gene together indicated that the virus from the man and the virus from the child were epidemiologically linked.

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Figures

FIG. 1

FIG. 1

Alignment of the amino acid sequences derived from the C2-V3-C3 region. The child's sequences correspond to the pt843t and pt843rt samples, and the sequences from the man correspond to the pt844t and v3prot165 samples. An asterisk indicates the described deletion between the child and the man. A minus sign indicates an amino acid that corresponds to the consensus sequence. Deletions in the amino acid alignment are indicated by a period. “X” corresponds to an amino acid mixture.

FIG. 2

FIG. 2

Phylogenetic tree derived from sequences from the C2-V3-C3 region. A total of 101 subtype B C2-V3-C3 sequences obtained from the Los Alamos database were included in the analyses, together with the 16 local control sequences (in boldface) and the four sequences from the man and the child (in boldface and underlined). The NJ method was used in constructing the tree. An HIV-1 subtype A sequence, U455, was used as an outgroup. The two lines (∥) on the outgroup indicate that the branch has been shortened.

FIG. 3

FIG. 3

(A) Phylogenetic tree derived from sequences from the p17gag region. (B) Phylogenetic tree derived from sequences from the C2-V3-C3 region. (C) Phylogenetic tree derived from sequences from the p17gag region and the C2-V3-C3 region combined. (D) Phylogenetic tree derived from sequences from the RT region of the pol gene. The NJ method was used in constructing all four trees. An HIV-1 subtype A sequence, U455, was used as an outgroup. The two lines (∥) on the outgroup indicate that the branch has been shortened. Only relevant bootstrap values have been included in the tree. Local controls are in boldface. The four sequences from the man and the child are in boldface and underlined.

FIG. 3

FIG. 3

(A) Phylogenetic tree derived from sequences from the p17gag region. (B) Phylogenetic tree derived from sequences from the C2-V3-C3 region. (C) Phylogenetic tree derived from sequences from the p17gag region and the C2-V3-C3 region combined. (D) Phylogenetic tree derived from sequences from the RT region of the pol gene. The NJ method was used in constructing all four trees. An HIV-1 subtype A sequence, U455, was used as an outgroup. The two lines (∥) on the outgroup indicate that the branch has been shortened. Only relevant bootstrap values have been included in the tree. Local controls are in boldface. The four sequences from the man and the child are in boldface and underlined.

FIG. 3

FIG. 3

(A) Phylogenetic tree derived from sequences from the p17gag region. (B) Phylogenetic tree derived from sequences from the C2-V3-C3 region. (C) Phylogenetic tree derived from sequences from the p17gag region and the C2-V3-C3 region combined. (D) Phylogenetic tree derived from sequences from the RT region of the pol gene. The NJ method was used in constructing all four trees. An HIV-1 subtype A sequence, U455, was used as an outgroup. The two lines (∥) on the outgroup indicate that the branch has been shortened. Only relevant bootstrap values have been included in the tree. Local controls are in boldface. The four sequences from the man and the child are in boldface and underlined.

FIG. 3

FIG. 3

(A) Phylogenetic tree derived from sequences from the p17gag region. (B) Phylogenetic tree derived from sequences from the C2-V3-C3 region. (C) Phylogenetic tree derived from sequences from the p17gag region and the C2-V3-C3 region combined. (D) Phylogenetic tree derived from sequences from the RT region of the pol gene. The NJ method was used in constructing all four trees. An HIV-1 subtype A sequence, U455, was used as an outgroup. The two lines (∥) on the outgroup indicate that the branch has been shortened. Only relevant bootstrap values have been included in the tree. Local controls are in boldface. The four sequences from the man and the child are in boldface and underlined.

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