Molecular Analyses
Concerning molecular detection of LCDV, the tested skin samples of nine subjects were negative.
Regarding bacteria identification, nine samples out of the 13 tested positive to the PCR targeting HSP65 gene of Mycobacterium sp. However, sequencing of the PCR products showed that not all of them were ascribable to the Mycobacterium genus. BLAST analysis of the obtained sequences identified mycobacteria in two samples (cases 9 and 11) and C. acnes in the other seven samples (cases 1, 2, 3, 4, 5, 6, and 8). In particular sequences obtained from samples of cases 1, 2, 3, 4, 5, 6, and 8 showed ≥98.5% nucleotide identity withPropionibacterium acnes ATCC 11828 (Genbank accession number CP003084); whereas, sequences from samples of cases 9 and 11 showed the highest similarity with Mycobacterium chelonae (100% nucleotide identity with JX154110) and M. parascrofulaceum (98,4% nucleotide identity with AY337276) respectively.