An outbreak of Legionella micdadei pneumonia in transplant patients: evaluation, molecular epidemiology, and control.

PURPOSE: To describe a nosocomial outbreak of Legionella micdadei pneumonia in transplant patients and to characterize the source of the outbreak and the control measures utilized. SUBJECTS AND METHODS: We performed retrospective Legionella micdadei serologic testing to enhance case finding in transplant patients with pneumonia that lacked a documented microbial etiology, as well as prospective environmental surveillance of water sites and testing for Legionella in clinical specimens. RESULTS: During a 3-month period, 12 cases of Legionella micdadei pneumonia were identified either by culture or serologic testing among 38 renal and cardiac transplant patients. Legionella micdadei isolates from hot water sources were found by pulsed-field gel electrophoresis to have a DNA banding pattern that was identical to the isolates from the first 3 culture-positive cases and from 2 cases that occurred 16 months later. CONCLUSIONS: Hospitals caring for organ transplant recipients and other immunosuppressed patients must be aware of the possibility of environmental sources of outbreaks of Legionella infection. A first-line screen with the Legionella urine antigen test will identify Legionella pneumophila serogroup 1. However, specific cultures in outbreak situations should be considered to identify other Legionella pneumophila serotypes and the nonpneumophila Legionella species.

Molecular epidemiology of vancomycin-resistant enterococci from 6 hospitals in New York State.

BACKGROUND: Vancomycin resistance among enterococci is an emerging nosocomial problem. Consequently, it is important to understand the distribution of vancomycin-resistant enterococci (VRE) within and between hospitals to implement appropriate infection control measures. METHODS: In this study, 116 VRE isolates obtained from patients in 6 New York State hospitals were analyzed by antibiotic susceptibility testing, pulsed-field gel electrophoresis (PFGE) fingerprinting, plasmid profile analysis, vanA and vanB polymerase chain reaction, and DNA:DNA hybridization with vanA and vanB probes. RESULTS: PFGE and plasmid typing generally agreed, but plasmid profiles were more variable. These analyses revealed that genetic heterogeneity among isolates from within each of the 6 hospitals varied considerably. Among 23 Enterococcus faecium isolates from one hospital, there were only 3 PFGE types, and 20 isolates had the same type. However, in another hospital, each isolate was genetically distinct. Closely related strains were not found in separate hospitals. VRE strains with vanA genes and strains with vanB genes were found in 3 hospitals. Both plasmid and chromosomal carriage of these genes was detected. CONCLUSIONS: PFGE typing showed that nosocomial VRE transmission had occurred in some hospitals. However, there was no evidence for it in others. Neither was there evidence for intrahospital transmission or for emergence of an endemic strain. These observations demonstrate that it is important to evaluate genetic heterogeneity among VRE before implementation of infection control measures. PFGE is the method of choice for epidemiologic typing, but polymerase chain reaction, plasmid, and hybridization studies can provide important information concerning the presence and potential for transfer of vancomycin resistance genes.

Staphylococcus aureus with reduced susceptibility to vancomycin isolated from a patient with fatal bacteremia.

A Staphylococcus aureus isolate with reduced susceptibility to vancomycin was obtained from a dialysis patient with a fatal case of bacteremia. Comparison of the isolate with two methicillin-resistant S. aureus (MRSA) isolated obtained from the same patient 4 months earlier suggests that the S. aureus with reduced susceptibility to vancomycin emerged from the MRSA strain with which the patient was infected. Atypical phenotypic characteristics, including weak or negative latex-agglutination test results, weak or negative-slide coagulase test results, heterogeneous morphologic features, slow rate of growth, and vancomycin susceptibility (by disk diffusion test) were observed.

Comparison of restriction enzyme analysis, arbitrarily primed PCR, and protein profile analysis typing for epidemiologic investigation of an ongoing Clostridium difficile outbreak.

During an outbreak of diarrhea in a general hospital in 1992, 166 Clostridium difficile isolates from 102 patients were typed by restriction enzyme analysis (REA), arbitrarily primed PCR (AP-PCR), and protein profile analysis (PP) techniques. A total of 18 types and 5 subtypes were identified by REA, 32 types were identified by AP-PCR, and 9 types were identified by PP. Analysis of the data indicated the presence of a predominant strain among 76, 75, and 84% of the isolates by REA, AP-PCR, and PP, respectively. Subsequently, 45 C. difficile isolates which had been collected in 1990 from 33 patients in the same hospital following a significant increase in the number of cases of diarrhea caused by C. difficile were studied by REA, AP-PCR, and PP typing techniques. Thirteen types and one subtype were identified by REA, 12 types were identified by AP-PCR, and 5 types were identified by PP. As with the isolates from 1992, a dominant strain was identified. This strain was represented by 53, 64, and 70% of the total number of isolates when the strains were typed by REA, AP-PCR, and PP, respectively. Every isolate (210 of 211) from both 1990 and 1992 that was available for typing was typeable by all three methods. Furthermore, the same dominant strain was identified in both 1990 and 1992 by each method. This study demonstrates that each of the three typing methods can be useful in epidemiologic investigations of C. difficile outbreaks and that one strain can be dominant in an institution over a number of years.

Genetic analysis of multiple vancomycin-resistant Enterococcus isolates obtained serially from two long-term-care patients.

Fifty-eight vancomycin-resistant enterococcal isolates were obtained from two patients over 9 weeks. Numerous pulsed-field gel electrophoresis fingerprinting types were isolated from each patient. By PCR, all isolates were vanA+. However, many isolates from patient B were found to lack vanA by hybridization. Our results demonstrate the importance of examining multiple isolates, especially from patients who are at high risk of infection.

Vancomycin-resistant enterococci in stool specimens submitted for Clostridium difficile cytotoxin assay.

The prevalence of, and clinical risk factors associated with, vancomycin-resistant enterococcal colonization were investigated in patients suspected of having Clostridium difficile infection. Stools submitted for C difficile cytotoxin testing were screened for vancomycin-resistant enterococci (VRE). Isolates were speciated and characterized further by antibiotic susceptibility testing, DNA fingerprinting, and DNA:DNA hybridization for detection of specific vancomycin resistance genes. Of the 79 evaluable patients identified during a 3-month period, 16.5% were VRE-positive. The VRE isolates were genetically heterogeneous, although all carried the vanA gene. DNA fingerprinting data suggest that patient-to-patient transmission occurred, implicating colonized patients as potential reservoirs for VRE transmission. A positive C difficile cytotoxin assay and diabetes mellitus were the only identifiable risk factors associated with VRE colonization. Patients at risk for C difficile infection therefore may serve as reservoirs for VRE.

Polymicrobial gram-negative bacteremia associated with saline solution flush used with a needleless intravenous system.

BACKGROUND: During a 2-week period, seven cases of nosocomial polymicrobial gram-negative rod bacteremia occurred on a 39-bed medical and cardiac step-down unit. Combinations of Enterobacter cloacae (seven isolates), Klebsiella pneumoniae (five isolates), and Citrobacter freundii (two isolates) were isolated from blood cultures. METHODS: Concurrent and retrospective chart reviews were used to look for further cases and common exposures. Epidemiologic methods were used to refine determination of common exposure. Restriction enzyme DNA analysis was performed on the isolates. RESULTS: Concurrent and retrospective chart reviews revealed four additional possible cases during the same period. All case patients were exposed, through peripheral saline solution locks or central venous catheters, to saline solution "flush" from a central 0.9% saline solution bag and a needleless dispensing pin. Epidemiologic methods implicated probable extrinsic contamination of a single bag and pin used during a 24-48-hour period (Fisher's Exact Test, p < 0.002). There were no other common exposures. Restriction enzyme DNA analysis of the isolates further supported a common source for the outbreak. CONCLUSIONS: The introduction of needleless intravascular systems has been embraced for employee protection. Our report is the first to raise the question of patient safety with such systems. This outbreak highlights the inherent risks in rapid introduction of new technologies and points out the delicate balance among patient health, employee safety, and cost containment.

Nosocomial legionellosis associated with aspiration of nasogastric feedings diluted in tap water.

PROBLEM: Two cases of nosocomial legionellosis due to Legionella pneumophila serogroup 6 (Lp6) were identified in the intensive care unit. Both patients had a history of aspiration of nasogastric tube feedings, developed pulmonary infiltrates, had positive cultures for Lp6, had serological titer rises to Legionella, were treated, and recovered. METHOD: Isolates of Lp6 from the potable water system and patients were characterized by DNA restriction enzyme analyses using pulsed-field gel electrophoresis (PFGE). RESULTS: Water samples grew > 10(4) CFU/L of Lp6, and the same PFGE pattern was observed with the patient and water isolates. Potable water was used only for delivering medications and diluting feeding solutions given through the nasogastric tubes of the patients. Heat shock of the hot water system (140 degrees to 160 degrees F or 60 degrees to 70 degrees C, 4 hours) was performed and the temperature was maintained between 131 degrees to 140 degrees F (55 degrees to 60 degrees C). Surveillance over 18 months revealed a reduction in Legionella to < 10(2) CFU/L. CONCLUSION: We speculate that nosocomial Legionella pneumonia occurred due to aspiration of nasogastric tube solutions diluted with tap water. A nursing practice change to use only sterile water to dilute feedings and flush medications for nasogastric administration was instituted. The hot water temperature at the faucet was increased to > or = 131 degrees F (> or = 60 degrees C) to control Legionella. No further nosocomial cases have occurred.

Comparison of ribotyping and restriction enzyme analysis using pulsed-field gel electrophoresis for distinguishing Legionella pneumophila isolates obtained during a nosocomial outbreak.

Because of the ubiquity of Legionella isolates in aquatic habitats, epidemiologic evaluation of Legionella pneumophila strains is important in the investigation and subsequent control of nosocomial outbreaks of legionellosis. In this study, ribotyping and restriction enzyme analysis by pulsed-field gel electrophoresis (PFGE) were used to compare isolates of L. pneumophila obtained from patients and the environment during a nosocomial outbreak with unrelated control strains. Restriction enzyme analysis by PFGE resolved 14 different patterns among the L. pneumophila serogroup 1 and L. pneumophila serogroup 6 isolates involved in the study. Two of the patterns were observed in the three L. pneumophila serogroup 6 isolates from patients with confirmed nosocomial infections and environmental isolates from the potable water supply, which was, therefore, believed to be the source of the patients' infections. Three more patterns that were not present in isolates from patients with legionellosis were seen in isolates from the hospital environment, demonstrating the presence of multiple strains in the hospital environment. In the outbreak, one distinct pattern occurred among the L. pneumophila serogroup 1 isolates from patients with nosocomial infections, suggesting a common source; however, the source could not be determined. By comparison, ribotyping generated five patterns. However, some control strains of both L. pneumophila serogroups 1 and 6 possessed the same ribotypes as were present in the outbreak isolates. Both techniques were used successfully to subtype the isolates obtained during the investigation of the outbreak. Furthermore, restriction enzyme analysis by PFGE was useful for subdividing ribotypes and for distinguishing strains involved in the outbreak from epidemiologically unrelated strains.

Modified immunofluorescence test for the serologic diagnosis of gonorrhea (FGT-ABS).

Investigation of the source of false-positive reactions in the Fluorescent Gonococcal Test-Heated (FGT-H) implicated antibodies to antigens common to Neisseria gonorrhoeae and N. meningitidis. Absorption experiments proved that the fluorescence was due to both species-specific and common antigens and that in most cases the common antibodies could be successfully absorbed by a soluble protein extract of N. meningitidis. The value of such an absorption step was examined using sera from 500 women, 73 from bacteriologically confirmed cases of gonorrhea and 427 from bacteriologically and clinically negative cases. The absorption step reduced the false positivity rate by 58% without a significant change in sensitivity. The modified procedure, the Fluorescent Gonococcal Antibody Test-Absorbed (FGT-ABS), has a sensitivity similar to the Fluorescent Gonococcal Test-Heated (FGT-H) but has the advantage of higher specificity.