J M Rolain1, M N Mallet, P E Fournier, D Raoult. 1. Didier Raoult, Unité des Rickettsies, Faculté de Médecine, 27, Boulevard Jean Moulin, 13385 Marseille Cedex 5, France.
Abstract
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.
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.
Authors: Kathleen E Mach; Ruchika Mohan; Ellen Jo Baron; Mei-Chiung Shih; Vincent Gau; Pak Kin Wong; Joseph C Liao Journal: J Urol Date: 2010-11-12 Impact factor: 7.450
Authors: M J Espy; J R Uhl; L M Sloan; S P Buckwalter; M F Jones; E A Vetter; J D C Yao; N L Wengenack; J E Rosenblatt; F R Cockerill; T F Smith Journal: Clin Microbiol Rev Date: 2006-01 Impact factor: 26.132
Authors: Nathan G Schoepp; Eugenia M Khorosheva; Travis S Schlappi; Matthew S Curtis; Romney M Humphries; Janet A Hindler; Rustem F Ismagilov Journal: Angew Chem Int Ed Engl Date: 2016-06-30 Impact factor: 15.336
Authors: Plamen A Demirev; Nathan S Hagan; Miquel D Antoine; Jeffrey S Lin; Andrew B Feldman Journal: J Am Soc Mass Spectrom Date: 2013-04-09 Impact factor: 3.109
Authors: Linda M Weigel; David Sue; Pierre A Michel; Brandon Kitchel; Segaran P Pillai Journal: Antimicrob Agents Chemother Date: 2010-05-03 Impact factor: 5.191
Authors: Brian J Morrow; Darren Abbanat; Ellen Z Baum; Steven M Crespo-Carbone; Todd A Davies; Wenping He; Wenchi Shang; Anne Marie Queenan; A Simon Lynch Journal: Antimicrob Agents Chemother Date: 2011-09-12 Impact factor: 5.191
Authors: Nadya Andini; Anne Hu; Luming Zhou; Steven Cogill; Tza-Huei Wang; Carl T Wittwer; Samuel Yang Journal: Clin Chem Date: 2018-08-07 Impact factor: 8.327