Growth inhibition of Mycobacterium tuberculosis by polyoxyethylene stearate present in the BACTEC pyrazinamide susceptibility test

Abstract

We have previously found that approximately 3.5% of 428 clinical isolates of Mycobacterium tuberculosis yield uninterpretable results in the BACTEC pyrazinamide (PZA) susceptibility test system, because of inadequate growth. We tested the hypothesis that polyoxyethylene stearate (POES), the ingredient of the reconstituting fluid for the test, was the cause of this growth inhibition. A total of 15 isolates known for their previously uninterpretable results and 100 randomly chosen clinical isolates were tested in parallel both with and without POES. Repeat testing of the isolates with previously uninterpretable results yielded results in the presence of POES in only seven (47%). In the absence of POES, all gave interpretable results but one such result showed false resistance. For the other 100 clinical isolates, interpretable results were obtained with and without POES, but growth was enhanced in the absence of POES, especially in the PZA-susceptible strains. This was evidenced by a decreased time to attain a growth index of 200 in the control vial (4.9 days without POES versus 5.8 days with POES; P < 0.001) and a higher mean growth index ratio on the day of interpretation of the test (7.4% without POES versus 2.2% with POES; P < 0.001). However, the enhanced growth without POES led to 20 susceptible strains being misinterpreted as either resistant or borderline. We suggest that isolates of M. tuberculosis which yield uninterpretable results in the BACTEC PZA test system should be retested both with and without POES. If interpretable results indicating PZA resistance are obtained only in the absence of POES, the result should be confirmed by a pyrazinamidase assay or by the conventional proportion method. Routine omission of POES from the BACTEC test for all clinical strains is discouraged because of the unacceptably high false-resistance rates.