Subject(s)
Bacterial Proteins/genetics , Bacteriological Techniques , DNA, Bacterial/genetics , DNA-Directed RNA Polymerases/genetics , Mycobacterium tuberculosis/genetics , Tuberculosis, Ocular/diagnosis , Vitreous Body/microbiology , Antibiotics, Antitubercular/therapeutic use , Case-Control Studies , Drug Resistance, Bacterial/genetics , Genotype , Humans , Mutation , Mycobacterium tuberculosis/drug effects , Predictive Value of Tests , Reproducibility of Results , Rifampin/therapeutic use , Tuberculosis, Ocular/drug therapy , Tuberculosis, Ocular/microbiologyABSTRACT
PURPOSE: To compare 3 different molecular techniques to detect the Mycobacterium tuberculosis genome in vitreous fluid of eyes with multifocal serpiginoid choroiditis (MSC). DESIGN: Prospective, interventional case series. PARTICIPANTS: Eleven patients (11 eyes) with active MSC in at least 1 eye underwent diagnostic pars plana vitrectomy (PPV) between October 2012 and December 2013. METHODS: Vitreous fluid samples were subjected to multitargeted polymerase chain reaction (PCR) for a M. tuberculosis assay, the Gene Xpert MTB/RIF assay (Cepheid, Sunnyvale, CA), and a line probe assay (GenoType MTBDRplus; Hain Lifescience, GmbH, Nehren, Germany). The samples with positive results were subjected to rpoB gene sequencing to demonstrate rifampicin resistance. The clinical details, digital fundus imaging, and treatment details and outcomes also were noted. MAIN OUTCOME MEASURES: Detection of the M. tuberculosis genome and rifampicin resistance in the vitreous samples. RESULTS: Of the 11 eyes subjected to PPV, the multitargeted PCR results for tuberculosis were positive for 10 eyes, the MTBDRplus assay results were positive in 6 eyes, and the Gene Xpert MTB/RIF assay results were positive in 4 eyes. Rifampicin resistance was detected in 3 eyes by rpoB gene sequencing, in 3 eyes by the MTBDRplus assay, and in 1 eye by the Gene Xpert MTB/RIF assay. CONCLUSIONS: We detected the M. tuberculosis genome in the vitreous fluid of eyes with MSC using 3 different molecular techniques. Rifampicin resistance was detected for the first time in eyes with MSC.
Subject(s)
Choroiditis/microbiology , Genome, Bacterial/genetics , Mycobacterium tuberculosis/isolation & purification , Tuberculosis, Multidrug-Resistant/microbiology , Tuberculosis, Ocular/microbiology , Vitreous Body/microbiology , Adolescent , Adult , Antibiotics, Antitubercular/pharmacology , Antitubercular Agents/therapeutic use , Choroiditis/diagnosis , Choroiditis/drug therapy , Drug Resistance, Bacterial , Drug Therapy, Combination , Female , Fluorescein Angiography , Humans , Male , Middle Aged , Molecular Diagnostic Techniques , Multifocal Choroiditis , Mycobacterium tuberculosis/drug effects , Mycobacterium tuberculosis/genetics , Polymerase Chain Reaction , Prospective Studies , Rifampin/pharmacology , Tuberculosis, Multidrug-Resistant/diagnosis , Tuberculosis, Multidrug-Resistant/drug therapy , Tuberculosis, Ocular/diagnosis , Tuberculosis, Ocular/drug therapy , Vitrectomy , Young AdultSubject(s)
Antibiotics, Antitubercular/therapeutic use , Rifampin/pharmacology , Tuberculosis, Multidrug-Resistant/drug therapy , Tuberculosis, Ocular/drug therapy , Adult , DNA, Bacterial/analysis , Female , Follow-Up Studies , Humans , Mycobacterium tuberculosis/genetics , Tuberculosis, Multidrug-Resistant/diagnosis , Tuberculosis, Multidrug-Resistant/microbiology , Tuberculosis, Ocular/diagnosisSubject(s)
Drug Resistance, Bacterial , Mycobacterium tuberculosis/isolation & purification , Tuberculosis/diagnosis , Tuberculosis/microbiology , Uveitis/diagnosis , Uveitis/microbiology , Adolescent , Adult , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Female , Genes, Bacterial , Genotype , Genotyping Techniques , Humans , Male , Mycobacterium tuberculosis/genetics , Sequence Analysis, DNA , Young AdultABSTRACT
We report the first case of peritonitis caused by Roseomonas mucosa which led to technique failure in an adolescent patient with HIV receiving peritoneal dialysis. Identification of the causative organism by 16S rRNA gene sequencing and phylogenetic analysis is described.
Subject(s)
Cross Infection/diagnosis , Gram-Negative Bacterial Infections/diagnosis , HIV Infections/complications , Methylobacteriaceae/isolation & purification , Peritoneal Dialysis/adverse effects , Peritonitis/diagnosis , Adolescent , Bacterial Typing Techniques , Cluster Analysis , Cross Infection/microbiology , Cross Infection/pathology , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Gram-Negative Bacterial Infections/microbiology , Gram-Negative Bacterial Infections/pathology , Humans , Peritonitis/microbiology , Peritonitis/pathology , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Young AdultABSTRACT
OBJECTIVES: To characterize isolates of Klebsiella pneumoniae producing KPC carbapenemase (KPC-Kp) associated with an outbreak in a long-term acute care hospital (LTACH) in South Florida. METHODS: During 21 March to 20 April 2008, 241 K. pneumoniae isolates detected at Integrated Regional Laboratories (Ft. Lauderdale, FL) for which the ertapenem MICs were > or =4 mg/L were studied. PCR, cloning and sequence analysis were used to detect bla(KPC) and to characterize the beta-lactamase and outer membrane proteins (Omps). The expression level of KPC enzymes was studied by immunoblotting. Genetic relatedness of isolates was investigated with rep-PCR and PFGE. Clinical records of patients were investigated. RESULTS: Seven KPC-Kp strains were isolated from different patients located at a single LTACH, with a further three isolates being recovered from patients at different hospitals. All KPC-Kp isolates in patients from the LTACH and from one hospital patient were genetically related and shared PFGE patterns that clustered with known sequence type (ST) 258 strains. These strains were highly resistant to carbapenems (MICs > or = 32 mg/L) due to an increased level of KPC expression and loss of Omps. Rectal colonization was documented in all LTACH patients with KPC-Kp isolates. Treatment failures were common (crude mortality rate of 69%). Active surveillance and enhanced infection control practices terminated the KPC-Kp outbreak. CONCLUSIONS: The detection of KPC-Kp in an LTACH represents a serious infection control and therapeutic challenge in a new clinical setting. The speed at which the epidemic of KPC-Kp is spreading in our healthcare system mandates urgent action.
Subject(s)
Bacterial Proteins/biosynthesis , Cross Infection/epidemiology , Disease Outbreaks , Klebsiella Infections/epidemiology , Klebsiella pneumoniae/enzymology , Klebsiella pneumoniae/isolation & purification , beta-Lactamases/biosynthesis , Adult , Aged , Aged, 80 and over , Anti-Bacterial Agents/pharmacology , Bacterial Outer Membrane Proteins/genetics , Bacterial Proteins/classification , Bacterial Proteins/genetics , Bacterial Typing Techniques , Cluster Analysis , Cross Infection/microbiology , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Electrophoresis, Gel, Pulsed-Field , Ertapenem , Female , Florida/epidemiology , Hospitals , Humans , Immunoblotting , Intensive Care Units , Klebsiella Infections/microbiology , Klebsiella pneumoniae/genetics , Male , Microbial Sensitivity Tests , Molecular Epidemiology , Polymerase Chain Reaction , Sequence Analysis, DNA , beta-Lactamases/classification , beta-Lactamases/genetics , beta-Lactams/pharmacologyABSTRACT
BACKGROUND: In July 1999, a rare strain of multidrug-resistant Salmonella enterica serovar Senftenberg was isolated from the sputum of a trauma patient. Over a 6-year period (1999-2005) in northeast Florida, this Salmonella serovar spread to 66 other patients in 16 different healthcare facilities as a result of frequent transfers of patients among institutions. To our knowledge, this is the first outbreak of healthcare-associated infection and colonization with a fluoroquinolone-resistant strain of S. Senftenberg in the United States. OBJECTIVES: To investigate an outbreak of infection and colonization with an unusual strain of S. Senftenberg and assist with infection control measures. DESIGN: A case series, outbreak investigation, and microbiological study of all samples positive for S. Senftenberg on culture. SETTING: Cases of S. Senftenberg infection and colonization occurred in hospitals and long-term care facilities in 2 counties in northeast Florida. RESULTS: The affected patients were mostly elderly persons with multiple medical conditions. They were frequently transferred between healthcare facilities. This Salmonella serovar was capable of long-term colonization of chronically ill patients. All S. Senftenberg isolates tested shared a similar pulsed-field gel electrophoresis (PFGE) pattern. CONCLUSION: A prolonged outbreak of infection and colonization with multidrug-resistant S. Senftenberg was identified in several healthcare facilities throughout the Jacksonville, Florida, area and became established when infection control measures failed. The bacterial agent was capable of long-term colonization in chronically ill patients. Because the dispersal pattern of this strain suggested a breakdown of infection control practices, a multipronged intervention approach was undertaken that included intense education of personnel in the different institutions, interinstitutional cooperation, and transfer paperwork notification.
Subject(s)
Cross Infection/epidemiology , Disease Outbreaks , Drug Resistance, Multiple, Bacterial , Salmonella Infections/epidemiology , Salmonella enterica/isolation & purification , Adolescent , Adult , Aged , Aged, 80 and over , Child , Child, Preschool , Cross Infection/microbiology , Cross Infection/transmission , Disease Transmission, Infectious , Electrophoresis, Gel, Pulsed-Field/methods , Female , Florida/epidemiology , Humans , Infection Control/methods , Male , Middle Aged , Retrospective Studies , Salmonella Infections/microbiology , Salmonella Infections/transmissionABSTRACT
An outbreak of typhoid fever in Florida involving at least 16 persons during the winter of 1998-99 was investigated using case-control, environmental, and laboratory methods. The genomic profiles of Salmonella serovar Typhi (Salmonella Typhi) isolates from the 15 confirmed case subjects were identical. Consumption of fruit shakes made with frozen mamey, a tropical fruit, was significantly associated with illness (matched odds ratio, 7.6; 95% confidence interval, 1.4-81.4). Laboratory testing showed that the fruit was heavily contaminated with fecal coliforms; no Salmonella Typhi was isolated. The frozen mamey was prepared in plants in Guatemala and Honduras. No further cases occurred after the frozen product was recalled. As our nation's food sources become increasingly globalized, the risk of outbreaks of exotic diseases linked to contaminated imported food will increase. This outbreak highlights the need for new approaches to ensure the safety of our food supply.