Investigation of human parvovirus B19 occurrence and genetic variability in different leukaemia entities A.C. da Costa, I. Bendit, A.C.S. de Oliveira,

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Investigation of human parvovirus B19 occurrence and genetic variability in different leukaemia entities A.C. da Costa, I. Bendit, A.C.S. de Oliveira, E.G. Kallas, E.C. Sabino, S.S. Sanabani Clinical Microbiology and Infection Volume 19, Issue 1, Pages E31-E43 (January 2013) DOI: 10.1111/1469-0691.12058 Copyright © 2013 European Society of Clinical Infectious Diseases Terms and Conditions

FIG. 1 Neighbour-joining inferred rooted phylogeny of a partial human B19V non-structural protein 1(NS1) sequence. The black circle denotes variants detected in patients surveyed in this study. The star symbol indicates patients dual-infected with genotypes 1a and 3b, as later determined by strategies 1 and 2. Bootstrap replication frequencies ≥ 70% are indicated above nodes. Genotypes and subgenotypes are indicated at the corresponding nodes. The tree was rooted on Rhesus macaque parvovirus (GenBank AF221122). Branch lengths are drawn to scale. Clinical Microbiology and Infection 2013 19, E31-E43DOI: (10.1111/1469-0691.12058) Copyright © 2013 European Society of Clinical Infectious Diseases Terms and Conditions

FIG. 2 Neighbour-joining inferred rooted phylogeny of human B19V non-structural protein 1/viral capsid proteins 1 and 2 (NS1/VP1/VP2) sequence. The black circle denotes variants detected in patients with partial human B19V NS1 sequences. The star symbol indicates patients dually infected with genotype 3b, as later confirmed by strategy 2. The black triangle symbol indicates an isolate forming a unique branch in genotype 1 separate from 1a and 1b, which appeared in further analysis as a 1a/3b recombinant (see Fig. 3). Bootstrap replication frequencies ≥ 70% are indicated above nodes. Genotypes and subgenotypes are indicated at the corresponding nodes. The tree was rooted on Rhesus macaque parvovirus (GenBank AF221122). Branch lengths are drawn to scale. Clinical Microbiology and Infection 2013 19, E31-E43DOI: (10.1111/1469-0691.12058) Copyright © 2013 European Society of Clinical Infectious Diseases Terms and Conditions

FIG. 3 Exploratory tree analysis based on fragments between breakpoints as indicated by a bootscan plot of isolate 07BRSP7612. The neighbour-joining tree was established from the alignments of the non-structural protein 1 (NS1) fragment spanning nucleotide positions 949–1874 (a) and positions 1886–2160 (B19 isolate J35 GenBank accession no. AY386330) (b) for isolate 07BRSP7612 (indicated by black circle) and other B19V reference genotypes. Bootstrap replication frequencies ≥ 70% are indicated above nodes. Genotypes and subgenotypes are indicated at the corresponding nodes. The tree was rooted on Rhesus macaque parvovirus (GenBank AF221122). Branch lengths are drawn to scale. Clinical Microbiology and Infection 2013 19, E31-E43DOI: (10.1111/1469-0691.12058) Copyright © 2013 European Society of Clinical Infectious Diseases Terms and Conditions

FIG. 4 Neighbour-joining inferred rooted phylogeny of the genotype 3b fragment involved in the recombinant event as defined by bootscan (274 base pairs, nucleotides 1886–2160 of B19 isolate J35 (GenBank accession no. AY386330) and the non-recombinant genotype 3b fragment as defined by strategy 3 from the 07BRSP7612 isolate (indicated by black circle). Bootstrap replication frequencies ≥ 65% are indicated above nodes. Genotypes and subgenotypes are indicated at the corresponding nodes. The tree was rooted on simian parvovirus (GenBank accession no AF221122). Branch lengths are drawn to scale. Clinical Microbiology and Infection 2013 19, E31-E43DOI: (10.1111/1469-0691.12058) Copyright © 2013 European Society of Clinical Infectious Diseases Terms and Conditions

FIG. 5 Exploratory tree analysis based on fragments between breakpoints as indicated by a bootscan plot of isolate 07BRSP7612 complete coding non-structural protein 1/viral capsid proteins 1 and 2 (NS1/VP1/VP2) sequence. The neighbour-joining tree was established from the alignments spanning nucleotide positions 913–1925 (a), 1929–2127 (b), and positions 2144–5023 (c) (B19 isolate J35 GenBank accession no. AY386330). Isolate 07BRSP7612 is indicated by circle. Bootstrap replication frequencies ≥ 70% are indicated above nodes. Genotypes are indicated at the corresponding nodes. Branch lengths are drawn to scale. Clinical Microbiology and Infection 2013 19, E31-E43DOI: (10.1111/1469-0691.12058) Copyright © 2013 European Society of Clinical Infectious Diseases Terms and Conditions