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A 1.5 Hour Procedure for Identification of Enterococcus Species Directly from Blood Cultures
Published on: February 10, 2011
Cluster of Enterobacter cloacae pseudobacteremias associated with use of an agar slant blood culturing system
M L Pearson1, D A Pegues, L A Carson
1National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333.
Abstract:
From 1 February through 12 October 1990, 27 blood cultures processed at Shiprock Hospital were positive for Enterobacter cloacae; only 3 had been reported in the preceding 12 months. Twenty (74%) of the cultures were obtained from patients without clinical evidence of gram-negative septicemia. The increase in E. cloacae-positive blood cultures was temporally associated with the introduction of a new blood culturing system. To evaluate potential risk factors for an E. cloacae-positive blood culture (case-culture), we conducted a case-control study. Case-cultures were compared with 81 randomly selected cultures that were processed during the epidemic period and that were not positive for E. cloacae (controls). Because several factors suggested the possibility of pseudoinfection, we limited our analysis to the 20 blood cultures that appeared to be contaminants. Blood samples received in the laboratory during the midnight shift (5 of 20 [25%] versus 5 of 81 [6%]; odds ratio, 5.1; 95% confidence intervals, 1.01 to 24.6; P = 0.02) or present in the incubator with other E. cloacae-positive samples (17 of 20 [85%] versus 29 of 81 [36%]; odds ratio, 10.2, 95% confidence interval, 2.6 to 57.3; P < 0.001) were at increased risk for contamination. During mock experiments of the procedures for processing blood samples for culture, several breaks in aseptic technique and leakage from the blood culturing system were observed. Cultures of samples obtained from several environmental sites in the laboratory and the hand washings of two laboratory technicians grew E. cloacae. Plasmid and restriction enzyme analyses of E. cloacae isolates recovered from the patients' blood cultures, the two technicians' hand washings, and environmental sites in the laboratory indicated that all had identical plasmid profiles. Our findings suggest that the breaks in aseptic technique and the environmental contamination that occurred in association with the use of the new blood culturing system resulted in contamination of the blood cultures. This outbreak highlights the importance of routine environmental cleaning, periodic quality control assessments, and adherence to aseptic practices in clinical laboratories, particularly when new methods or equipment are introduced and/or new personnel are hired.
Insights
A new blood culturing system at Shiprock Hospital led to an Enterobacter cloacae outbreak. Breaks in aseptic technique and lab contamination caused pseudoinfections, highlighting the need for strict protocols.
Area of Science:
- Clinical Microbiology
- Infectious Diseases
- Laboratory Science
Background:
- An unusual increase in Enterobacter cloacae (E. cloacae) positive blood cultures was observed at Shiprock Hospital.
- The rise coincided with the implementation of a new blood culturing system.
Purpose of the Study:
- To investigate the cause of the E. cloacae outbreak.
- To identify risk factors associated with contaminated blood cultures.
Main Methods:
- A case-control study was conducted comparing E. cloacae positive cultures with negative cultures.
- Analysis focused on suspected pseudoinfections (cultures from patients without septicemia symptoms).
- Investigated aseptic technique, equipment integrity, and environmental contamination, including plasmid analysis of isolates.
Main Results:
- Blood samples processed during midnight shifts and incubated with other positive samples showed increased contamination risk.
- Breaks in aseptic technique and leakage from the new culturing system were identified.
- E. cloacae isolates from patients, technicians, and the environment shared identical plasmid profiles, indicating a common source.
Conclusions:
- The outbreak was attributed to environmental contamination and breaches in aseptic technique linked to the new blood culturing system.
- Emphasizes the critical importance of rigorous aseptic practices, quality control, and environmental monitoring in clinical laboratories, especially during system changes.

