Figure 1.
a) La Doncella (the Maiden); b) El Niño (the Boy); and c) La Niña (the Girl).
Table 1.
Primers used for PCR amplification from DNA extracted from swab samples.
Figure 2.
Gel electrophoresis from the Maiden.
Gel electrophoresis (2%) showing the amplification of the Mycobacterium sp. hsp65 gene fragments in the Maiden’s buccal swab samples 6 and 11, both sequenced successfully. The image was inverted to facilitate detection of bands and no other image treatment was performed. Sample 16 did not produce a satisfactory sequence. Samples 3 and 4 were positive for primers described as specific to Mycobacterium avium, but did not produce satisfactory sequences. Sample 1 and 7 were positive for primers described as specific to Mycobacterium tuberculosis, but did not produce satisfactory sequences. The negative controls are in lanes 17, 18, 19 and 20. Ladders are 100-bp Fisher exACTgene.
Figure 3.
Gel electrophoresis from the boy.
Gel electrophoresis (2%) showing the products of PCR amplification from samples from the boy. The results were negative: no amplification bands could be detected. The image was inverted, and contrast was increased to facilitate detection of bands. No other image treatment was performed. Lanes 1 and 2 were amplified with M. tuberculosis-specific primers, lanes 3 and 4 with the Mycobacterium-specific primers, and lanes 5, 6 and 7 were negative controls.
Figure 4.
Maximum likelihood phylogeny and bootstrap support values based on 1000 pseudoreplicates of the alignment of hsp65 gene nucleotide sequences. Sequences generated from the Maiden’s swab samples are shown in bold, larger font, and marked with an asterisk.
Figure 5.
Alternative phylogenies compared to the best phylogeny.
Alternative phylogenies compared to the best phylogeny obtained through maximum likelihood analyses of DNA sequence data from the Maiden’s swab samples (fig. 4) using Bayesian posterior probabilities, approximately unbiased, and weighted Shimodaira-Hasegawa tests. See table 2 for description of each alternative and test results.
Table 2.
Maximum likelihood, Bayesian posterior probability (BPP), and significance of alternative phylogenies (for numbering, see fig. 5) using the approximately unbiased (AU) and weighted Shimodaira-Hasegawa (WSH) tests.
Table 3.
Immune system proteins and their respective accession numbers in the swab sample of the Maiden identified by mass spectrometry.