Antibiotic Susceptibility Testing of Grown Blood Cultures by Combining Culture and Real-Time Polymerase Chain Reaction Is Rapid and Effective

Beuving, Judith; Verbon, Annelies; Gronthoud, Firza Alexander; Stobberingh, Ellen E.; Wolffs, Petra

doi:10.1371/journal.pone.0027689

Cited by 31 publications

(36 citation statements)

References 35 publications

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“…The advantage of this method is the possibility of measuring bacteria in the range of 1 to 6 ϫ 10 4 cell counts; however, compared to the oCelloScope system, an AST can be performed for only one antibiotic at a time, and no image material that can reveal any erroneous results is available. The principles of several other methods of performing rapid ASTs have been presented (17)(18)(19)(20)(21)(22)31); a common limitation of these techniques is their inability to perform multiple-sample analysis.…”

Section: Discussionmentioning

confidence: 99%

“…Presently, these techniques are limited by single-sample analysis and the requirement for specialized technical personnel. Molecular methods, such as real-time PCR (22), mass spectrometry (23), microarrays (24), and flow cytometry (25), have been developed for rapid bacterial identification and ASTs because of their high sensitivity and promptness. A wide range of bacteria can be identified within minutes using mass spectrometry (26), and flow cytometry predicts antimicrobial susceptibility within 90 to 120 min (25).…”

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confidence: 99%

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Real-Time Optical Antimicrobial Susceptibility Testing

et al. 2013

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Rapid antibiotic susceptibility testing is in high demand in health care fields as antimicrobial-resistant bacterial strains emerge and spread. Here, we describe an optical screening system (oCelloScope) which, based on time-lapse imaging of 96 bacteria-antibiotic combinations at a time, introduces real-time detection of bacterial growth and antimicrobial susceptibility with imaging material to support the automatically generated graphs. Automated antibiotic susceptibility tests of a monoculture showed statistically significant antibiotic effects within 6 min and within 30 min in complex samples from pigs suffering from catheter-associated urinary tract infections. The oCelloScope system provides a fast high-throughput screening method for detecting bacterial susceptibility that might entail an earlier diagnosis and introduction of appropriate targeted therapy and thus combat the threat from multidrug-resistant pathogenic bacteria. The oCelloScope system can be employed for a broad range of applications within bacteriology and might present new vistas as a point-of-care instrument in clinical and veterinary settings.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Real-Time Optical Antimicrobial Susceptibility Testing

et al. 2013

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“…For AST, a semi-molecular method called FAST, previously described by our group [14], was used for Staphylococcus aureus, Enterococcus species and all GNRs, but not for streptococci, since they require different culture conditions.…”

Section: Standard Of Care (Soc)mentioning

confidence: 99%

“…Recently, we developed a new protocol for same-day AST, called FAST [14]. This method uses highly sensitive polymerase chain reaction (PCR) to detect bacterial growth in the presence of antibiotics after only 6 h. The results are, thus, based on culture-based antibiotic susceptibility profiles and were shown to be as reliable as most commonly used automated AST systems.…”

Section: Introductionmentioning

confidence: 99%

Impact of same-day antibiotic susceptibility testing on time to appropriate antibiotic treatment of patients with bacteraemia: a randomised controlled trial

Beuving

Wolffs

Hansen

et al. 2014

Eur J Clin Microbiol Infect Dis

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Inadequate therapy in bloodstream infections is suggested to be associated with higher mortality. We evaluated the reduction in inappropriate antibiotic therapy using rapid identification and antibiotic susceptibility testing (FAST) compared to standard of care (SOC) testing in patients with bloodstream infections. The FAST method used polymerase chain reaction (PCR) for identification and to detect growth in the presence or absence of antibiotics after only 6 h. For SOC testing, the BD Phoenix system was used. Patients with blood cultures growing Staphylococcus, Streptococcus or Enterococcus species or Gram-negative rods were randomised for FAST or SOC tests. A total of 129 patients were randomised for FAST and 121 patients for the SOC group. At the time SOC results became available, 78 patients in the FAST group could have been switched to more appropriate therapy. Although FAST results were highly accurate (agreement with SOC was 94%), they were only implemented in a minority (16) of patients. However, significantly fewer patients in the FAST group used inappropriate therapy at the time of SOC results (p = 0.025). The time to results in the FAST group was reduced by 15.6 h (p < 0.001). In the patients switched after FAST, this was done after a mean of 42.3 h compared to 61.4 h in those switched after SOC tests (p < 0.001). In bacteraemic patients, FAST resulted in significantly more patients using appropriate antibiotic therapy at the time SOC results were available and 15.6 h earlier than SOC tests. However, the implementation of FAST results was not optimal and no benefit on clinical outcome was shown.

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“…and (facultative) aerobe Gram-negative rods 6 . This assay was based on a study in which PCR was used to measure the growth of bacteria 7 .…”

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One-day Workflow Scheme for Bacterial Pathogen Detection and Antimicrobial Resistance Testing from Blood Cultures

Hansen

Beuving

Verbon

et al. 2012

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Bloodstream infections are associated with high mortality rates because of the probable manifestation of sepsis, severe sepsis and septic shock 1 . Therefore, rapid administration of adequate antibiotic therapy is of foremost importance in the treatment of bloodstream infections. The critical element in this process is timing, heavily dependent on the results of bacterial identification and antibiotic susceptibility testing. Both of these parameters are routinely obtained by culture-based testing, which is time-consuming and takes on average 24-48 hours 2,4 . The aim of the study was to develop DNA-based assays for rapid identification of bloodstream infections, as well as rapid antimicrobial susceptibility testing. The first assay is a eubacterial 16S rDNA-based real-time PCR assay complemented with species-or genus-specific probes 5 . Using these probes, Gram-negative bacteria including Pseudomonas spp., Pseudomonas aeruginosa and Escherichia coli as well as Gram-positive bacteria including Staphylococcus spp., Staphylococcus aureus, Enterococcus spp., Streptococcus spp., and Streptococcus pneumoniae could be distinguished. Using this multiprobe assay, a first identification of the causative micro-organism was given after 2 h.Secondly, we developed a semi-molecular assay for antibiotic susceptibility testing of S. aureus, Enterococcus spp. and (facultative) aerobe Gram-negative rods 6 . This assay was based on a study in which PCR was used to measure the growth of bacteria 7 . Bacteria harvested directly from blood cultures are incubated for 6 h with a selection of antibiotics, and following a Sybr Green-based real-time PCR assay determines inhibition of growth. The combination of these two methods could direct the choice of a suitable antibiotic therapy on the same day (Figure 1).In conclusion, molecular analysis of both identification and antibiotic susceptibility offers a faster alternative for pathogen detection and could improve the diagnosis of bloodstream infections.

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Antibiotic Susceptibility Testing of Grown Blood Cultures by Combining Culture and Real-Time Polymerase Chain Reaction Is Rapid and Effective

Cited by 31 publications

References 35 publications

Real-Time Optical Antimicrobial Susceptibility Testing

Real-Time Optical Antimicrobial Susceptibility Testing

Impact of same-day antibiotic susceptibility testing on time to appropriate antibiotic treatment of patients with bacteraemia: a randomised controlled trial

One-day Workflow Scheme for Bacterial Pathogen Detection and Antimicrobial Resistance Testing from Blood Cultures

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