Real-time PCR for universal antibiotic susceptibility testing

Objectives: Determination of bacterial antimicrobial susceptibility is usually performed using phenotypic methods. In this study, we developed a universal 16S rRNA and rpoB quantitative PCR assay for susceptibility testing of bacteria commonly isolated in clinical microbiology laboratories. Methods:...

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Bibliographic Details
Published in:Journal of antimicrobial chemotherapy 2004-08, Vol.54 (2), p.538-541
Main Authors: Rolain, J. M., Mallet, M. N., Fournier, P. E., Raoult, D.
Format: Article
Language:English
Subjects:
antibiotic resistance
Antibiotics. Antiinfectious agents. Antiparasitic agents
Bacteria - drug effects
Bacteria - growth & development
Bacterial Infections - microbiology
Biological and medical sciences
Computer Systems
DNA, Bacterial - chemistry
DNA, Bacterial - genetics
DNA-Directed RNA Polymerases - genetics
Humans
Kinetics
Medical sciences
Microbial Sensitivity Tests - methods
MICs
Pharmacology. Drug treatments
Phenotype
quantitative PCR
Reverse Transcriptase Polymerase Chain Reaction - methods
RNA, Bacterial - genetics
RNA, Ribosomal, 16S - genetics
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Summary:Objectives: Determination of bacterial antimicrobial susceptibility is usually performed using phenotypic methods. In this study, we developed a universal 16S rRNA and rpoB quantitative PCR assay for susceptibility testing of bacteria commonly isolated in clinical microbiology laboratories. Methods: Antibiotic susceptibilities for 24 bacterial strains of various species were tested by real-time quantitative PCR assay and by conventional methods. Quantification of DNA copies of either the 16S RNA genes or rpoB were recorded over time in the presence or absence of antibiotics to determine the bacterial growth kinetics and the optimal testing time. Results: Molecular results for antibiotic susceptibility or resistance were in accordance with those obtained using a standard macrodilution broth assay. The method was reproducible, sensitive and rapid (2 h for Gram-negative bacilli and 4 h for Gram-positive cocci). Moreover, this assay was also able to determine the antibiotic susceptibilities of fastidious bacteria, such as mycobacteria, within 5 days. Conclusions: These results demonstrate that molecular detection of bacteria could be more rapid than phenotypic methods for antibiotic susceptibility testing.
ISSN:0305-7453
1460-2091
1460-2091
DOI:10.1093/jac/dkh324