Comparison of three techniques for concentrating positive BACTEC 13A bottles for mycobacterial DNA probe analysis

Three methods of concentrating 1 ml aliquots from BACTEC 13A bottles containing patient blood samples were evaluated for testing with the Gen-Probe Rapid Diagnostic System for Mycobacteria avium complex: 1. using no reagents, 2. using both lysing and wash reagents; and 3. using lysing reagent only. Aliquots from 13As containing human blood and seeded with eight mycobacterial species were also concentrated directly and using both reagents. Results for samples containing M. avium were as follows: 1. using the direct concentration technique 34 of 47 samples (72%) gave unequivocally positive results; 2. 43 of 47 samples (92%) concentrated using both reagents gave positive results; 3. the technique using lysing reagent only was not found useful. There were no false positives with any of the seeded specimens. We were also able to define the minimum Growth Index necessary to ensure un-equivocally positive results for each concentration technique. For those samples containing M. avium these values were 42 for the technique using both reagents and 86 for the direct technique. Direct or reagent concentration of 13A aliquots for testing with Gen-Probe DNA probes provides a rapid, sensitive, and specific means for the identification of M. avium complex bacteremia.