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Yip, C.W. et al.

Denaturing HPLC for high-throughput screening of rifampicin-resistant Mycobacterium tuberculosis isolates.

OBJECTIVE: To evaluate the use of denaturation high-performance liquid chromatography (dHPLC) as a rapid method to detect rifampicin (RMP) resistance based on mutations in the rpoB gene in a high-volume laboratory setting. METHODS: A total of 132 RMP-resistant Mycobacterium tuberculosis strains with different rpoB mutation were used to optimise the running condition of dHPLC as a pilot study. A blind correlation study was subsequently done between dHPLC and in vitro RMP susceptibility tests on 3167 M. tuberculosis strains in a high-throughput clinical setting. RESULTS: In the pilot study, rpoB mutation could be detected on 116/132 (87.9%) RMP-resistant strains by dHPLC. In the second phase of the study, 84/3107 (2.7%) clinical M. tuberculosis isolates were RMP-resistant. The sensitivity and specificity of dHPLC in the prediction of RMP resistance were 70/84 (83.3%) and 70/77 (91.0%), respectively. The specificity became 100% when 511 Leu to Pro mutation was excluded from the RMP resistance-related genetic changes. CONCLUSION: In the detection of RMP resistance in a high-throughput laboratory setting, dHPLC has been demonstrated to be rapid, simple, workable, automatable and inexpensive in terms of running costs and the labour involved.[1]

References

Denaturing HPLC for high-throughput screening of rifampicin-resistant Mycobacterium tuberculosis isolates. Yip, C.W., Leung, K.L., Wong, D., Cheung, D.T., Chu, M.Y., Tang, H.S., Kam, K.M. Int. J. Tuberc. Lung Dis. (2006) [Pubmed]

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