ABSTRACT
The global emergence of carbapenem-resistant Enterobacteriaceae (CRE) presents a significant clinical concern, prompting the WHO to prioritize CRE as a top priority pathogen in their 2017 global antibiotic-resistant bacteria priority list. Due to the fast-depleting antibiotic arsenal, clinicians are now resorting to using once-abandoned, highly toxic antibiotics such as the polymyxins and aminoglycosides, creating an urgent need for new antibiotics. Drug repurposing, the application of an approved drug for a new therapeutic indication, is deemed a plausible solution to this problem. A total of 1,163 FDA-approved drugs were screened for activity against a clinical carbapenem- and multidrug-resistant E. coli isolate using a single-point 10 µM assay. Hit compounds were then assessed for their suitability for repurposing. The lead candidate was then tested against a panel of clinical CREs, a bactericidal/static determination assay, a time-kill assay and a checkerboard assay to evaluate its suitability for use in combination with Tigecycline against CRE infections. Three drugs were identified. The lead candidate was determined to be Zidovudine (azidothymidine/AZT), an oral anti-viral drug used for HIV treatment. Zidovudine was shown to be the most promising candidate for use in combination with Tigecycline to treat systemic CRE infections. Further experiments should involve the use of animal infection models.
Subject(s)
Anti-Bacterial Agents/therapeutic use , Carbapenem-Resistant Enterobacteriaceae/drug effects , Drug Repositioning , Enterobacteriaceae Infections/drug therapy , Escherichia coli/drug effects , Minocycline/analogs & derivatives , Zidovudine/therapeutic use , Animals , Drug Resistance, Multiple, Bacterial , Enterobacteriaceae Infections/microbiology , Humans , Mice , Microbial Sensitivity Tests , Minocycline/therapeutic use , TigecyclineABSTRACT
Current autoclaving practices are designed to kill bacteria. Little is known about the effect of autoclaving on the integrity of bacterial genomic DNA. An experiment was performed to examine the effect of standard autoclaving on the integrity of bacterial DNA, employing polymerase chain reaction (PCR) as an indicator of DNA integrity. Amplifiable PCR signal was observed at t = 10, 20 and 30 min autoclaving time for Pseudomonas aeruginosa NCTC 10662; at t = 10, 20, 30 and 40 min for Salmonella Nottingham NCTC 7832; and at t = 10 and 20 min for Escherichia coli NCTC 9001. Careful consideration should therefore be given to residual molecular artefacts in future risk and environmental impact assessments, where the legacy of residual genomic DNA from dead bacterial and higher organisms may act as a potential reservoir, thereby feeding horizontal gene transfer scenarios in viable cells with potential hazardous genes of virulence, persistence or antibiotic resistance characteristics.
Subject(s)
DNA, Bacterial/radiation effects , Escherichia coli/genetics , Gene Transfer, Horizontal/radiation effects , Medical Waste Disposal/methods , Pseudomonas aeruginosa/genetics , Salmonella/genetics , Sterilization/methods , DNA, Bacterial/genetics , Escherichia coli/radiation effects , Hot Temperature , Humans , Polymerase Chain Reaction , Pseudomonas aeruginosa/radiation effects , Salmonella/radiation effects , Time FactorsABSTRACT
The use of fibreoptic bronchoscopy with sterile catheter sampling of pulmonary secretions was evaluated in 70 patients with a provisional diagnosis of pneumonia. In 37 patients quantitative analysis of the sterile catheter isolates was performed (colony forming units (CFU) per ml). Potential bacterial pathogens were isolated in 37 patients and in the quantitative analysis, 14 of 22 isolates were grown in counts greater than or equal to 10(3) CFU/ml. Sterile catheter increased the bacterial isolation rate as in only 19 patients blood (2) or sputum (18) cultures yielded the same organisms. Sputum cultures showed a 25% false-positive rate in patients with no growth from sterile catheter. Quantitative analysis did not yield any further information in patients receiving antibiotics. Atypical or fungal pneumonia was diagnosed in 22 patients, while ten patients had other pathology simulating pneumonia. Sterile catheter sampling of pulmonary secretions at fibreoptic bronchoscopy proved to be a valuable tool in the diagnosis of bacterial pneumonia.
