Mycobacterium abscessus infection in solid organ transplant recipients: report of three cases and review of the literature.

Mycobacterium abscessus is an ubiquitous organism found in the environment. This rapidly growing mycobacterium infrequently causes disease in humans; however, in immunocompromised hosts, disease can range from localized cutaneous lesions to disseminated infection. The organism is resistant to most antimycobacterial drugs and therapy can be limited by drug interactions. The exact incidence of M. abscessus infection among solid organ transplant (SOT) recipients is unknown; data are only available from previously reported cases in the literature. We describe 3 cases of M. abscessus infection in SOT recipients diagnosed within a 5-month period. One of the cases followed multi-visceral transplantation, the first such case to be reported in the literature. An epidemiological investigation did not reveal significant commonalities among the cases, and pulsed-field gel electrophoresis of genomic DNA of the case isolates confirmed their non-identity. All cases improved with antibiotic therapy, most notably with the new glycylcycline, tigecycline, along with surgical intervention in 2 of the cases. In addition, we review features and characteristics of M. abscessus infections in recipients of SOT reported in the literature from 1992 to 2008 and summarize some selected therapeutic concerns and issues related to treatment.

Increased bloodstream infection rates in surgical patients associated with variation from recommended use and care following implementation of a needleless device.

OBJECTIVE: To determine if an apparent increase in bloodstream infections (BSIs) in patients with central venous catheters (CVCs) was associated with the implementation of a needleless access device. DESIGN: Retrospective cohort study using a derived CVC-days factor for estimating appropriate denominator data. SETTING: A 350-bed urban, acute, tertiary-care hospital. METHODS: BSI surveillance data were obtained, and high-risk areas for BSIs were determined. A random 5% sample of medical records was used to estimate CVC days, and a cohort study was conducted to compare BSI rates before and during needleless device use. A survey was conducted of nursing needleless-device practices. RESULTS: The surgical intensive-care unit (SICU), the medical intensive-care unit, and the solid organ transplant unit (OTU) were identified as high-risk units. Using existing surveillance BSI data and the estimated CVC days, the catheter-related BSI rates in the high-risk surgical patients were significantly higher during the needleless-device period compared with the preneedleless-device period (SICU, 9.4 vs 5.0/1,000 CVC days; OTU, 13.6 vs 2.2/1,000 CVC days). A survey of the nurses revealed that 60% to 70% were maintaining the needleless devices correctly. CONCLUSION: We observed a significant increase in the BSI rate in two surgical units, SICU and OTU, associated with introduction of a needleless device. This increase occurred shortly after the needleless device was implemented and was associated with nurses' unfamiliarity with the device, and needless-device use and care practices different from the manufacturer's recommendations.

Use of an estimation method to derive an appropriate denominator to calculate central venous catheter-associated bloodstream infection rates.

An outbreak investigation was conducted to determine if an increase in bloodstream infections (BSIs) in patients with central venous catheters (CVC) had occurred. Because other methods of obtaining CVC days were not feasible, we used an estimation method based on a random 5% sample of medical records to determine the proportion of days that a CVC was present for each of three patient units. This calculated ratio was used to estimate the total CVC days for each unit. A cohort study was conducted in which the BSI rates before and during needleless device use were compared. This article describes the methods used to calculate this estimated denominator and discusses the need for such a denominator to be used by infection control practitioners when prospective collection of CVC days is not possible.

Evaluation of the acceptability of a needleless vascular-access system by nurses.

BACKGROUND: Needleless intravenous-access devices have been introduced in an effort to reduce needlestick injuries and possible transmission of blood-borne pathogens to health care workers. However, there are no data on the acceptance of these devices by nursing personnel. METHODS: A survey of nursing personnel was taken at Indiana University Medical Center after introduction of a needleless intravenous device to determine their opinion after use of the needleless device. RESULTS: The majority of the nurses (72 of 94, 70%) had a favorable overall opinion of the device. Among those with a favorable opinion, 76% (55/72) responded that reduced risk of needlestick injury was the most important reason. Among those who had a negative opinion about the needleless-device system, 32% (7/22) reported that contamination risk was their major concern. Those who were trained before device use were more likely to properly use and maintain the needleless intravenous-access system. Of 89 respondents, 75.3% (67/89) believed that the initial training was adequate; however, 43% (29/67) thought that additional training after using the device for some time would have been beneficial. CONCLUSIONS: Comprehensive education programs that include training before and after device use are necessary if new needleless intravenous-access systems are to be successfully introduced and accepted by nursing personnel.

Decrease in expenditures and selected nosocomial infections following implementation of an antimicrobial-prescribing improvement program.

OBJECTIVE: To evaluate changes in antimicrobial use and expenditures and the rates of selected nosocomial infections due to resistant organisms associated with implementation of an antimicrobial-prescribing improvement program. DESIGN: Before-after trial comparing 1992 (pre-program), 1993 (a transition year), and 1994 (after full implementation of the program). SETTING AND PARTICIPANTS: Academic medical center, all patients and physicians. INTERVENTION: An antimicrobial-prescribing improvement program with prior approval requirement for use of restricted agents. MAIN OUTCOME MEASURES: Antimicrobial use and expenditures, rates of selected nosocomial infection marker events. RESULTS: Between 1992 and 1994, there were substantial decreases in antimicrobial use, from 158,107 to 137,364 defined daily doses, and in expenditures from $2,486,902 ($24.01 per patient day) to $1,701,522 ($18.49 per patient day). After adjusting for changes in purchase prices and census days, we estimated savings attributable to the program of $279,573 in 1993 and $389,814 in 1994. In addition, we found significant decreases between 1992 and 1994 in the rates of enterococcal bacteremia (.34 vs .16 events per 1,000 patient days; P = .016), selected gram-negative bacteremia (.26 vs .11; P = .015), methicillin-resistant Staphylococcus aureus colonization or infection (.66 vs .20; P < .0001), and Stenotrophomonas colonization or infection (.35 vs .17; P = .019). No significant change occurred in rates of nosocomial candidemia or Clostridium difficile toxin-positive diarrhea. Values for 1993 were intermediate between those of 1992 and 1994. CONCLUSION: Implementation of an antimicrobial-prescribing improvement program was associated with substantial savings in antimicrobial use and expenditures and significant decreases in rates of selected nosocomial infections due to resistant organisms.

DNA typing and control of methicillin-resistant Staphylococcus aureus at two affiliated hospitals.

OBJECTIVE: To describe control of endemic and outbreak-related methicillin-resistant Staphylococcus aureus (MRSA) at two affiliated hospitals. DESIGN: Prospective surveillance of patients with MRSA. Disposable gloves were used by all staff having direct contact with the affected patient or his immediate environment, and patient isolates were typed by pulsed-field gel electrophoresis (PFGE) of genomic DNA. Surveillance and PFGE typing were used concurrently to identify possible nosocomial outbreaks, confirm or refute cross-infection, and support a need for additional outbreak control interventions. SETTING: A university hospital (Hospital A) and a university-affiliated public hospital (Hospital B). PARTICIPANTS: Patients with MRSA colonization or infection over an 18-month interval (June 1993-November 1994). INTERVENTION: Proper handwashing and gloving practices were reemphasized with staff following confirmation of outbreaks. RESULTS: Hospital A had 60 community-acquired and 48 nosocomial cases of MRSA. Two small outbreaks (affecting a total of seven patients) and two pseudo-outbreaks were identified. Hospital B had 36 community-acquired and 22 nosocomial cases of MRSA. Only one outbreak affecting five patients occurred. All outbreaks ended shortly after staff meetings that emphasized ongoing and extremely careful handwashing and gloving when caring for identified patients. The majority of nosocomial cases at both hospitals were not related epidemiologically or had isolates with unique PFGE types. Pseudo-outbreaks were confirmed by demonstrating that isolates from epidemiologically related cases (by time and clinical service or hospital unit) had different PFGE types. Hospital A cases had 39 different PFGE types, and Hospital B cases had 31 different PFGE types. CONCLUSION: MRSA in hospitals, including outbreaks identified by prospective surveillance and confirmed by PFGE typing, can be controlled by minimal special precautions and interventions. This is possible despite the continuous admission of patients with MRSA from the community. PFGE typing is useful to confirm outbreaks and pseudo-outbreaks, demonstrate differences among epidemiologically unrelated isolates, and substantiate the efficacy of MRSA control programs within hospitals.

Characterization of DFA-negative, probe-positive Neisseria gonorrhoeae by pulsed field gel electrophoresis.

Eight Neisseria gonorrhoeae isolates, negative by direct fluorescent antibody (DFA) but positive by a DNA probe, were characterized by pulsed field gel electrophoresis and compared to eight DFA-positive, probe-positive isolates. Results indicate that DFA-negative, probe-positive Neisseria gonorrhoeae isolates may be clonal.

In vivo stability and discriminatory power of methicillin-resistant Staphylococcus aureus typing by restriction endonuclease analysis of plasmid DNA compared with those of other molecular methods.

We evaluated test discriminatory power and DNA type alterations among methicillin-resistant Staphylococcus aureus strains by testing 199 sequential isolates from 39 patients collected over 30 to 228 days. Isolates were typed by one or three different methods (restriction endonuclease analysis of plasmid DNA [REAP] with or without pulsed-field gel electrophoresis of genomic DNA [PFGE] and immunoblotting [IB]). REAP was highly discriminatory compared with PFGE and IB. However, the initial isolates from 4 of the 39 patients lacked detectable plasmid DNA and could not be typed by REAP. Typing of individual patient isolates showed that a different REAP type was identified only once every 138 days. Among 25 comparisons, seven sequential isolate pairs demonstrating REAP differences were also different by PFGE and IB. This likely represented the presence of more than one strain. Eighteen other pairs with REAP differences were identical or related to one another by PFGE and IB typing, and 17 of these differences were likely caused by a single genetic alteration within the same strain or clone. The rate of PFGE differences explicable by single genetic alterations among sequential isolates identical by REAP was similar to the overall rate for REAP differences in the whole collection. We conclude that REAP and PFGE typing differences explicable by single genetic alterations are relatively infrequent but not rare. These isolates should be examined by alternative typing systems to further support or refute clonality.

Typing of sequential bacterial isolates by pulsed-field gel electrophoresis.

We typed 39 sets of multiple bacterial isolates of the same species from patients by pulsed-field gel electrophoresis of genomic DNA (PFGE). Isolates were cultured from different sites or over a 2-week or longer interval. Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, and Enterobacter cloacae were tested. Excluding E. cloacae, 28 of 32 sets of isolates (87%) demonstrated only identical or highly related PFGE types. Four of the seven sets of E. cloacae showed different types. For species other than E. cloacae, our results suggest that patients are usually colonized and infected with a single strain of these bacterial pathogens. Unlike all of the other tested species, E. cloacae PFGE typing differences suggested the presence of multiple strains causing colonization and infection.

Control of methicillin-resistant Staphylococcus aureus in a hospital and an intensive care unit.

OBJECTIVE: To describe methicillin-resistant Staphylococcus aureus (MRSA) control in a hospital, including a surgical intensive care unit (SICU) outbreak. DESIGN: Prospective surveillance of newly identified patients with MRSA. Barrier isolation (disposable gloves for direct contact with patient or immediate environment) was used for the routine care of hospitalized MRSA patients as of October 1991. Beginning in 1992, MRSA isolates were typed by restriction endonuclease enzyme analysis of plasmid DNA (REAP) and/or pulsed-field gel electrophoresis of genomic DNA (PFGE). Surveillance information and MRSA typing were used concurrently to identify nosocomial case clustering, confirm cross-infection, and support a need for additional outbreak control interventions. SETTING: University-affiliated public hospital. PARTICIPANTS: Patients with newly identified MRSA colonization or infection from 1991 through 1993 and epidemiologically associated staff providing care to eight SICU patients in an outbreak. INTERVENTIONS: Barrier isolation for affected and unaffected patients in and admitted to the SICU institution when the outbreak was identified and cross-infection confirmed. Anterior nares cultures of staff in contact with outbreak cases for detection of MRSA colonization. RESULTS: Fifty-six hospitalized patients with community-acquired MRSA and 80 patients with nosocomial MRSA colonization or infection were identified during the 3 years. After the introduction of barrier isolation, the annual frequency of new nosocomial MRSA cases decreased and only one outbreak (eight cases in the SICU) caused by type-related isolates occurred. The other 35 nosocomial cases of MRSA during 1992 and 1993 were not epidemiologically related or were caused by isolates with different types. The SICU outbreak ended after instituting barrier isolation for all patients (with and without MRSA) in and admitted to the unit. Six colonized SICU staff were identified. All outbreak cases had identical or related MRSA types by PFGE and REAP. Staff isolates were different from case isolates by typing, and staff were not restricted and not given treatment for colonization. After more than 6 months of follow up, no further outbreaks of MRSA in the SICU or elsewhere in the hospital occurred despite returning to barrier isolation for affected patients only. CONCLUSION: MRSA in hospitals and outbreaks of MRSA in ICUs can be controlled by surveillance and minimal barrier interventions. REAP or PFGE typing of MRSA can be used to support or refute the presence of cross-transmission. Typing also may be helpful when planning and assessing the effectiveness of interventions directed at endemic, as well as outbreak, MRSA control.

Assessment of bacterial cross-transmission as a cause of infections in patients in intensive care units.

The prevalence of possible cross-transmission of selected bacteria (Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Pseudomonas aeruginosa, Staphylococcus aureus, and enterococci) among infected patients was evaluated in five intensive care units (ICUs) over 6 months. A total of 284 isolates from clinical specimens were typed by plasmid profile analysis (E. coli, K. pneumoniae, and E. cloacae), restriction endonuclease analysis of plasmid DNA (S. aureus), and/or pulse-field gel electrophoresis of chromosomal DNA (P. aeruginosa, enterococci, S. aureus, and other bacteria without plasmid DNA). By typing criteria, only 13% of the 177 isolates obtained after > 2 days in an ICU were classified as possibly cross-transmitted. Many patients whose cultures yielded bacteria of an identical type may have been the sources rather than the recipients of these organisms. Episodes of possible cross-transmission were scattered among all ICUs, usually affected only two patients, and were associated with most bacterial species. These data suggest that endemic bacterial cross-transmission in ICUs is relatively infrequent and that cross-transmitted bacteria are not common causes of endemic ICU-related nosocomial infections.

Value of molecular epidemiologic analysis in a nosocomial methicillin-resistant Staphylococcus aureus outbreak.

OBJECTIVE: To evaluate two molecular epidemiologic methods used in the analysis of a nosocomial methicillin-resistant Staphylococcus aureus (MRSA) outbreak. DESIGN: Restriction endonuclease analysis of plasmid DNA (REAP) was used in the analysis of 45 MRSA isolates. After termination of the outbreak, isolates were retrospectively analyzed in a blind fashion using the newly described technique of arbitrarily primed polymerase chain reaction (AP-PCR). Molecular analyses were compared with epidemiologic and antimicrobial susceptibility data. SETTING: Tertiary care university hospital. SUBJECTS: Twenty-eight patients and 12 employees infected or colonized with MRSA during a 6-week period. RESULTS: A clonal relationship demonstrated among isolates from burn unit patients and staff was clearly distinguishable from MRSA isolates arising from other hospital wards. The combination of REAP and AP-PCR provided complementary information in several instances. Aggressive measures to isolate infected patients and eradicate colonization from patients and staff terminated the outbreak. CONCLUSIONS: Although traditional epidemiologic methods retain their central role in modern hospital infection control, molecular epidemiologic analysis can significantly enhance the ability of infection control officers to analyze and terminate hospital epidemics. The combination of AP-PCR and REAP may prove to be a particularly informative means of tracking the nosocomial spread of microbial strains and their mobile genetic elements.

Plasmid DNA analysis, biotyping, and antimicrobic susceptibility as subtyping tests for Klebsiella pneumoniae and Klebsiella oxytoca.

We compared plasmid DNA analysis, biotyping by Vitek, and disk diffusion antimicrobic susceptibility as subtyping tests of Klebsiella pneumoniae and Klebsiella oxytoca. The 92 tested isolates were from alternate, culture-positive patients over 6 months. No outbreak or cluster of infections was recognized during this interval. Plasmid DNA was detected in 85% of the isolates. Each isolate except one had a reproducible absence of plasmid DNA or a reproducible plasmid DNA profile on repetitive testing. Restriction endonuclease enzyme analysis of plasmid DNA was necessary to distinguish differences among some isolates that had only large plasmids. Isolates with only large plasmids represented 18% of the collection. Of the 78 isolates with plasmid DNA, all but two were considered different from one another by plasmid DNA analysis. Biotyping and antimicrobic susceptibility testing were not highly reproducible. In addition, biotyping did not demonstrate a sufficient variety of patterns among the isolates for subtyping purposes. We conclude that plasmid DNA analysis is very useful as a subtyping test for isolates of K. pneumoniae and K. oxytoca. Neither biotyping nor antimicrobial susceptibility as performed in our laboratory had sufficient discriminatory power and reproducibility for subtyping these organisms.

Molecular epidemiology of Candida albicans colonization and fungemia in very low birthweight infants.

OBJECTIVE: This study investigated the relationship between colonization and fungemia. DESIGN: This was a prospective study involving surveillance cultures of the nares, base of umbilicus, point of entry of umbilical catheter and parenteral fluids. Blood cultures were done when sepsis was suspected. All Candida albicans isolates were typed using restriction enzyme analysis of DNA. SETTING: Patients were from the neonatal intensive care unit of a tertiary care hospital. POPULATION STUDIED: Twenty-nine very low birthweight infants. MAIN RESULTS: Eleven babies were colonized with C albicans and five of these babies developed fungemia, including five of seven who were colonized at the point of entry of the umbilical catheter. Three different strains of C albicans caused fungemia. In four of the five patients, initial catheter entry site isolates were identical to the subsequent blood isolates. Occasionally, infants were colonized with more than one strain of C albicans. CONCLUSIONS: Preceding colonization with C albicans and, in particular, colonization at the site of entry of umbilical vascular catheters are risk factors for subsequent development of C albicans fungemia. Fungemic and colonizing isolates are usually identical to one another by DNA typing.

Recurrent Staphylococcus aureus bacteremia.

Sequential blood isolates from eight patients with 10 episodes of recurrent Staphylococcus aureus bacteremia were typed by restriction endonuclease analysis of plasmid DNA (REAP DNA fingerprinting) and immunoblotting. There were six early recurrences (within 2 months of stopping antimicrobial therapy) and four late recurrences. All early recurrences isolates were identical to initial isolates. These recurrences were defined as possible relapses. Three of four late recurrence isolates were different from the preceding isolates recovered from four patients. This was considered indicative of new infections. There was complete concordance between REAP DNA fingerprinting and immunoblot typing results. However, four isolates lacked plasmid DNA and could be typed only by immunoblotting. All initial isolates from different patients were different types by immunoblotting and by REAP DNA fingerprinting (except for those lacking plasmid DNA). The bacterial traits detected by these methods appear to be stable in vivo for up to 3 months. Relapsing infections were associated with the presence of intravascular foreign bodies and vancomycin therapy of the preceding episodes.

Reproducibility of Staphylococcus epidermidis plasmid profiles.

The plasmid profiles of six isolates of Staphylococcus epidermidis were repetitively evaluated over an 8-month period. Each isolate was subcultured and stored at three different temperatures (-70 degrees C, -20 degrees C, and room temperature) and plasmid DNA was prepared from each subculture at 0, 1, 4, and 8 months by two different methods of plasmid extraction [using mixed alkyltrimethylammonium bromide (ATAB) or Brij 58 and deoxycholate (modified Parisi)]. Plasmid DNA bands were lost from two isolates when subcultures were kept at room temperature. This plasmid loss was confirmed by repetitive extractions and electrophoresis, as well as by restriction endonuclease analysis of the ATAB preparations. Profiles were otherwise highly related to one another, with occasional exceptions being extra or missing plasmid DNA bands of high molecular size. The latter findings were not reproducible. Plasmid DNA extracted by the modified Parisi method was not reliably digested with restriction endonuclease enzymes. We conclude that the plasmid profiles of Staphylococcus epidermidis isolates are highly reproducible as long as isolates are stored at less than or equal to -20 degrees C. Minor discrepancies in the number of plasmid DNA bands of large molecular size may occur. These are resolvable by repetitive testing or restriction endonuclease analysis of ATAB-extracted plasmid DNA preparations.