Rapid microarray-based assay for detection of pyrazinamide resistant Mycobacterium tuberculosis

Juliane Havlicek; Beatrice Dachsel; Peter Slickers; Sönke Andres; Patrick Beckert; Silke Feuerriegel; Stefan Niemann; Matthias Merker; Ines Labugger

doi:10.1016/j.diagmicrobio.2018.12.011

Rapid microarray-based assay for detection of pyrazinamide resistant Mycobacterium tuberculosis

Juliane Havlicek, Beatrice Dachsel, Peter Slickers, Sönke Andres, Patrick Beckert, Silke Feuerriegel, Stefan Niemann, Matthias Merker^*, Ines Labugger

^*Corresponding author for this work

Clinic of Internal Medicine III - Pulmonology

Abstract

Pyrazinamide (PZA)is a key antibiotic for the treatment of drug susceptible tuberculosis. PZA-resistance is mainly mediated by mutations in the pncA gene; however the current gold standard is a phenotypic drug susceptibility test requiring a well-adjusted pH-value for reliable results. Our melting curve assay detects a non-wild type genotype in selected pncA regions in at least 3750 gene copies/mL within 2.5 hours. The prototype assay was further evaluated by analyzing 271 Mycobacterium tuberculosis complex isolates from Swaziland originating from a previously published drug resistance survey and including 118 isolates with pncA mutations. Sensitivity was 83% (95% CI 75–89%)and specificity was 100% (95% CI 98–100%). Under consideration of further improvements with regard to the target range our melting curve assay has the potential as a rapid rule-in test for PZA susceptibility (wild type pncA), however false resistant results (mutant pncA, but PZA susceptible)cannot be ruled out completely.

Original language	English
Journal	Diagnostic Microbiology and Infectious Disease
Volume	94
Issue number	2
Pages (from-to)	147-154
Number of pages	8
ISSN	0732-8893
DOIs	https://doi.org/10.1016/j.diagmicrobio.2018.12.011
Publication status	Published - 06.2019

Funding

This work was supported by a grant from the Bill & Melinda Gates Foundation (grant number OPP1068618 ).

Research Areas and Centers

Academic Focus: Center for Infection and Inflammation Research (ZIEL)

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.diagmicrobio.2018.12.011

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title = "Rapid microarray-based assay for detection of pyrazinamide resistant Mycobacterium tuberculosis",

abstract = "Pyrazinamide (PZA)is a key antibiotic for the treatment of drug susceptible tuberculosis. PZA-resistance is mainly mediated by mutations in the pncA gene; however the current gold standard is a phenotypic drug susceptibility test requiring a well-adjusted pH-value for reliable results. Our melting curve assay detects a non-wild type genotype in selected pncA regions in at least 3750 gene copies/mL within 2.5 hours. The prototype assay was further evaluated by analyzing 271 Mycobacterium tuberculosis complex isolates from Swaziland originating from a previously published drug resistance survey and including 118 isolates with pncA mutations. Sensitivity was 83\% (95\% CI 75–89\%)and specificity was 100\% (95\% CI 98–100\%). Under consideration of further improvements with regard to the target range our melting curve assay has the potential as a rapid rule-in test for PZA susceptibility (wild type pncA), however false resistant results (mutant pncA, but PZA susceptible)cannot be ruled out completely.",

author = "Juliane Havlicek and Beatrice Dachsel and Peter Slickers and S{\"o}nke Andres and Patrick Beckert and Silke Feuerriegel and Stefan Niemann and Matthias Merker and Ines Labugger",

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year = "2019",

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doi = "10.1016/j.diagmicrobio.2018.12.011",

language = "English",

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AU - Havlicek, Juliane

AU - Dachsel, Beatrice

AU - Slickers, Peter

AU - Andres, Sönke

AU - Beckert, Patrick

AU - Feuerriegel, Silke

AU - Niemann, Stefan

AU - Merker, Matthias

AU - Labugger, Ines

N1 - Funding Information: This work was supported by a grant from the Bill & Melinda Gates Foundation (grant number OPP1068618 ). Publisher Copyright: © 2018 The Author(s) Copyright: Copyright 2019 Elsevier B.V., All rights reserved.

PY - 2019/6

Y1 - 2019/6

N2 - Pyrazinamide (PZA)is a key antibiotic for the treatment of drug susceptible tuberculosis. PZA-resistance is mainly mediated by mutations in the pncA gene; however the current gold standard is a phenotypic drug susceptibility test requiring a well-adjusted pH-value for reliable results. Our melting curve assay detects a non-wild type genotype in selected pncA regions in at least 3750 gene copies/mL within 2.5 hours. The prototype assay was further evaluated by analyzing 271 Mycobacterium tuberculosis complex isolates from Swaziland originating from a previously published drug resistance survey and including 118 isolates with pncA mutations. Sensitivity was 83% (95% CI 75–89%)and specificity was 100% (95% CI 98–100%). Under consideration of further improvements with regard to the target range our melting curve assay has the potential as a rapid rule-in test for PZA susceptibility (wild type pncA), however false resistant results (mutant pncA, but PZA susceptible)cannot be ruled out completely.

AB - Pyrazinamide (PZA)is a key antibiotic for the treatment of drug susceptible tuberculosis. PZA-resistance is mainly mediated by mutations in the pncA gene; however the current gold standard is a phenotypic drug susceptibility test requiring a well-adjusted pH-value for reliable results. Our melting curve assay detects a non-wild type genotype in selected pncA regions in at least 3750 gene copies/mL within 2.5 hours. The prototype assay was further evaluated by analyzing 271 Mycobacterium tuberculosis complex isolates from Swaziland originating from a previously published drug resistance survey and including 118 isolates with pncA mutations. Sensitivity was 83% (95% CI 75–89%)and specificity was 100% (95% CI 98–100%). Under consideration of further improvements with regard to the target range our melting curve assay has the potential as a rapid rule-in test for PZA susceptibility (wild type pncA), however false resistant results (mutant pncA, but PZA susceptible)cannot be ruled out completely.

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M3 - Journal articles

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SN - 0732-8893

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JO - Diagnostic Microbiology and Infectious Disease

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IS - 2

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Rapid microarray-based assay for detection of pyrazinamide resistant Mycobacterium tuberculosis

Abstract

Funding

Research Areas and Centers

UN SDGs

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