Rapid Pyrazinamide Drug Susceptibility Testing using a Closed-Tube PCR Assay of the Entire pncA gene

The continued use of pyrazinamide in the treatment of tuberculosis in the absence of a rapid, accurate and standardized pyrazinamide drug susceptibility assays is of great concern. While whole genome sequencing holds promise, it is not yet feasible option in low resource settings as it requires expe...

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Published in:	Scientific reports 2020-03, Vol.10 (1), p.4234-4234, Article 4234
Main Authors:	Whitfield, Michael G, Marras, Salvatore A E, Warren, Rob M, Van Rie, Annelies, Rice, John, Wangh, Lawrence J, Kreiswirth, Barry N
Format:	Article
Language:	eng
Subjects:	Amidohydrolases - genetics Antitubercular Agents - pharmacology Genomes Humans Microbial Sensitivity Tests - methods Mutation Mycobacterium tuberculosis Mycobacterium tuberculosis - drug effects Mycobacterium tuberculosis - genetics Phenotype PncA gene Polymerase Chain Reaction - methods Pyrazinamide Pyrazinamide - pharmacology Tuberculosis Whole genome sequencing
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Summary:	The continued use of pyrazinamide in the treatment of tuberculosis in the absence of a rapid, accurate and standardized pyrazinamide drug susceptibility assays is of great concern. While whole genome sequencing holds promise, it is not yet feasible option in low resource settings as it requires expensive instruments and bioinformatic analysis. We investigated the diagnostic performance of a closed-tube Linear-After-The-Exponential (LATE)-PCR assay for pyrazinamide susceptibility in Mycobacterium tuberculosis. Based on a set of 654 clinical Mycobacterium tuberculosis culture isolates with known mutations throughout the pncA gene as determined by Sanger sequencing, the assay displays excellent sensitivity of 96.9% (95% CI: 95.2-98.6) and specificity of 97.9% (95% CI: 96.1-99.7). In a subset of 384 isolates with phenotypic drug susceptibility testing, we also observed high sensitivity of 98.9% (95% CI: 97.5-100) but lower specificity of 91.8% (95% CI: 87.9-95.8) when compared to phenotypic drug susceptibility testing. We conclude that the LATE PCR assay offers both a rapid and accurate prediction of pyrazinamide susceptibility.
ISSN:	2045-2322 2045-2322