TY - JOUR
T1 - Rapid microarray-based assay for detection of pyrazinamide resistant Mycobacterium tuberculosis
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.
UR - http://www.scopus.com/inward/record.url?scp=85060943765&partnerID=8YFLogxK
U2 - 10.1016/j.diagmicrobio.2018.12.011
DO - 10.1016/j.diagmicrobio.2018.12.011
M3 - Journal articles
C2 - 30733004
AN - SCOPUS:85060943765
SN - 0732-8893
VL - 94
SP - 147
EP - 154
JO - Diagnostic Microbiology and Infectious Disease
JF - Diagnostic Microbiology and Infectious Disease
IS - 2
ER -
