Multi-Probe Real-Time PCR Identification of Common Mycobacterium Species in Blood Culture Broth Suporn Foongladda, Suporn Pholwat, Boonchuay Eampokalap,

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Multi-Probe Real-Time PCR Identification of Common Mycobacterium Species in Blood Culture Broth Suporn Foongladda, Suporn Pholwat, Boonchuay Eampokalap, Pattarachai Kiratisin, Ruengpung Sutthent The Journal of Molecular Diagnostics Volume 11, Issue 1, Pages 42-48 (January 2009) DOI: 10.2353/jmoldx.2009.080081 Copyright © 2009 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Alignment of 16S rRNA gene fragments of various mycobacterial species from the NCBI database and the clinical isolates. Nucleotides corresponding to the primers are in bolds. Sequences for the upstream and downstream hybridization probes are in shadows. The first consensus nucleotide corresponds to E. coli 16S rRNA gene (GenBank accession number M25051) position 55. Dot indicates identity and dash indicates deletion from the above consensus sequence. The Journal of Molecular Diagnostics 2009 11, 42-48DOI: (10.2353/jmoldx.2009.080081) Copyright © 2009 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Real-time PCR amplification curves and the melting curves at channel 705 (A), 640 (B), and 610 (C) detections: 1. M. tuberculosis ATCC 27294; 2. M. avium ATCC 700898; 3. M. intracellulare ATCC 13950; 4. M. kansasii ATCC 12478; 5. M. scrofulaceum ATCC19981; 6. M. simiae ATCC 25275; 7. M. sherrisii S24; 8. M. terrae ATCC 15750; 9. M. szulgai ATCC 35799; 10. M. chelonae S12; 11. M. abscessus S16; 12. M. xenopi ATCC 19250; 13. M. gordonae ATCC 14470; and 14. negative control. The Journal of Molecular Diagnostics 2009 11, 42-48DOI: (10.2353/jmoldx.2009.080081) Copyright © 2009 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 Real-time PCR amplification curves and melting curves channel 705-M. tuberculosis (A) and channel 640- M. avium (B) in the same reaction tube. 1. M. tuberculosis positive control; 2. M. avium positive control; 3. through 13., samples; and 14. negative control. Sample numbers 3 to 10 were identified as M. tuberculosis complex. Sample numbers 11 to 13 were identified as M. avium. The Journal of Molecular Diagnostics 2009 11, 42-48DOI: (10.2353/jmoldx.2009.080081) Copyright © 2009 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions