Speed of molecular detection techniques for methicillin-resistant Staphylococcus aureus admission screening in an acute care hospital.

Active surveillance for methicillin-resistant Staphylococcus aureus (MRSA) carriers is considered an essential component of MRSA control strategies in acute care hospitals. Recently, molecular assays for MRSA screening have been proposed with significant reduction of the sample processing time. Using a time analysis model, we investigated the time gain after the introduction of a molecular assay and compared this with a preceding control period, using culture-based techniques. During a four-month period all high risk patients (N=44) and all known MRSA-positive patients readmitted to the hospital (N=41) were screened for MRSA upon admission. In both groups the long pre-analytical phase - time from admission to sampling and transportation of samples to the laboratory - was the determining factor in the entire process. A substantial reduction of the sample processing time was achieved using molecular assays, compared with conventional culture. Due to the long pre-analytical phase, in addition to the high costs associated with polymerase chain reaction (PCR) testing, molecular techniques were not introduced for the admission screenings. In the group of the readmission screenings, however, a fast test result could save a substantial number of unnecessary isolation days, resulting in an economic benefit for the hospital. PCR testing might be of interest for the readmission screenings. In conclusion, local policies for MRSA screening should be investigated before introducing expensive PCR technology.

Role of coagulase-negative staphylococci in human disease.

Coagulase-negative staphylococci (CNS) are normal inhabitants of human skin and mucous membranes. They have long been dismissed as culture contaminants, but now the potentially important role of CNS as pathogens and their increasing incidence has been recognized. Approximately 55-75% of nosocomial isolates is methicillin resistant. CNS were the first organisms in which glycopeptide resistance was recognized. In the immunocompetent host, CNS endocarditis and urinary tract infections with Staphylococcus saprophyticus are the most common CNS infections. Other patients are usually immunocompromised, with indwelling or implanted foreign bodies. CNS account for approximately 30% of all nosocomial blood stream infections. The majority of these concern catheter-related sepsis. Other important infections due to CNS include central nervous system shunt infections, endophthalmitis, surgical site infections, peritonitis in patients with continuous ambulatory peritoneal dialysis and foreign body infections. CNS are rarely associated with mastitis in humans. Staphylococcus lugdunensis is more pathogenic than other CNS as it expresses several potential virulence factors. The distinction between clinically significant, pathogenic and contaminating isolates is a major problem. Several studies show clonal intra and inter hospital spread of Staphylococcus epidermidis strains which suggests that infection control measures may be necessary for multiresistant CNS isolates as for methicillin resistant Staphylococcus aureus. As a result of medical progress, mainly due to the use of invasive and indwelling medical devices, CNS are now a major cause of nosocomial and health-care related infections.

Costs and length of stay associated with antimicrobial resistance in acute kidney injury patients with bloodstream infection.

INTRODUCTION: Antimicrobial resistance negatively impacts on prognosis. Intensive care unit (ICU) patients, and particularly those with acute kidney injury (AKI), are at high risk for developing nosocomial bloodstream infections (BSI) due to multi-drug-resistant strains. Economic implications in terms of costs and length of stay (LOS) attributable to antimicrobial resistance are underevaluated. This study aimed to assess whether microbial susceptibility patterns affect costs and LOS in a well-defined cohort of ICU patients with AKI undergoing renal replacement therapy (RRT) who developed nosocomial BSI. METHODS: Historical study (1995-2004) enrolling all adult RRT-dependent ICU patients with AKI and nosocomial BSI. Costs were considered as invoiced in the Belgian reimbursement system, and LOS was used as a surrogate marker for hospital resource allocation. RESULTS: Of the 1330 patients with AKI undergoing RRT, 92 had microbiologic evidence of nosocomial BSI (57/92, 62% due to a multi-drug-resistant microorganism). Main patient characteristics were equal in both groups. As compared to patients with antimicro-4 bial-susceptible BSI, patients with antimicrobial-resistant BSI were more likely to acquire Gram-positive infection (72.6% vs 25.5%, P<0.001). No differences were found neither in LOS (ICU before BSI, ICU, hospital before BSI, hospital, hospital after BSI, and time on RRT; all P>0.05) or hospital costs (all P>0.05) when comparing patients with antimicrobial-resistant vs antimicrobial-susceptible BSI. However, although not statistically significant, patients with BSI caused by resistant Gram-negative-, Candida-, or anaerobic bacteria incurred substantial higher costs than those without. CONCLUSION: In a cohort of ICU patients with AKI and nosocomial BSI undergoing RRT, patients with antimicrobial-resistant vs antimicrobial-susceptible Gram-positive BSI did not have longer hospital stays, or higher hospital costs. Patients with resistant "other" (i.e. Gram-negative, Candida, or anaerobic) BSI were found to have a distinct trend towards increased resources use as compared to patients with susceptible "other" BSI, respectively.

Systematic surveillance cultures as a tool to predict involvement of multidrug antibiotic resistant bacteria in ventilator-associated pneumonia.

OBJECTIVE: To assess prediction of multidrug resistant (MDR) pathogens in ventilator-associated pneumonia (VAP) by systematic surveillance cultures (SC) and to assess the contribution of SC to initial antibiotic therapy. DESIGN: Prospective cohort study of patients with microbiologically confirmed VAP. Comparison of actual early antibiotic coverage with three hypothetical empirical schemes. SETTING: A 50-bed university hospital ICU. SC consisted of oral, nasal, urinary and rectal samples upon admission, 3-weekly urinary and 1-weekly oral, nasal, and rectal samples in all patients, 3-weekly tracheal aspirates in intubated patients. RESULTS: MDR pathogens were found in 86 of 199 VAP episodes. Sensitivity of SC to predict MDR pathogens was 69% (tracheal SC) and 82% (all SC); specificity was 96% (tracheal) and 91% (all), respectively. Appropriate antibiotic coverage within 24 h and 48 h following MDR VAP was 77% and 89%, respectively. A carbapenem-based empirical scheme would have been equally appropriate (83% vs. 77% at 24 h; 83% vs. 89% at 48 h), but a beta-lactam-fluoroquinolone empirical therapy would have been less (59% vs. 77% at 24 h; 59% vs. 89% at 48 h) as would have been beta-lactam-aminoglycoside therapy (68% vs. 77% at 24 h; 68% vs. 89% at 48 h). Empirical comparators would have resulted in significantly more prescription of broad-spectrum antibiotics within the first 48 h. CONCLUSIONS: With MDR pathogens highly prevalent, systematic SC predicted MDR pathogens causing VAP in 69% to 82% and may have contributed to high rates of early appropriate antibiotic therapy with limited use of broad-spectrum antimicrobials.

Towards a Belgian consensus for prevention of perinatal group B streptococcal disease.

BACKGROUND & OBJECTIVES: In Belgium, as in many other countries, group B Streptococcus (GBS) is still the leading cause of sepsis and meningitis in neonates. In 2001, though no Belgian guidelines for their prevention were available, in some hospitals, obstetrical programmes included a GBS prevention policy. With an aim to reach a Belgian consensus for the prevention of perinatal group B streptococcal disease, a national consensum meeting was organized in 2001. We report here our experience and findings of this meeting. METHODS: In November 2001, obstetricians, neonatologists, microbiologists and infectious diseases specialists were invited to participate in a GBS symposium. International and Belgian speakers presented epidemiological aspects, argued comparative cost-effectiveness of different approaches for prevention and debated technical and practical problems. Management of neonates with risk factors for GBS disease and progress in GBS vaccines were also included in the programme. Further results about Belgian obstetricians' practice and compliance to a policy for prevention of neonatal GBS diseases, as answered in two mail surveys, were commented and discussed. In an interactive session at the end, each participant was asked to vote on the key points related to the different steps of the ideal prevention strategy to recommend. RESULTS: For the main questions, 94 per cent of participants choose a screening-based approach and 94 per cent shifted from the current use of ampicillin to penicillin as first choice for antimicrobial prophylaxis. Further, 79 per cent voted for an approach with integrated neonatal prophylaxis for selected neonates at high risk for GBS disease and 47 per cent voted for a strategy based on an intrapartum rapid screening-based approach. INTERPRETATION & CONCLUSION: The state of the question by different speakers, the data from Belgian epidemiology, and the debate about cost-effectiveness of different approaches led to a massive vote in favour of the universal screening-based approach. Based on these results, a working group has been appointed by the Ministry of Health to draft and edit Belgian recommendations for the prevention of perinatal GBS disease.

Ten years phage-typing of Belgian clinical methicillin-resistant Staphylococcus aureus isolates (1992-2001).

Gradual changes have been observed in the phage-types of methicillin-resistant Staphylococcus aureus (MRSA) isolates from Belgian hospitals. A total of 6551 isolates, collected in 93 Belgian hospitals over 10 years (1992-2001), was examined. A decreasing incidence of the main early Belgian epidemic phage-types ([A], [B], [H]*, Jo*) was observed. Since 1997 and 2000, a new series of phage-types ([Hv]*, [J]*, [O]*), which were likely related to the previous group [H]*, have been noted. The general trends were confirmed in two particular hospitals. Local epidemic and/or endemic phage-types were also encountered.

Urinary infections in patients with spinal cord injury.

STUDY DESIGN: A retrospective study concerning urinary tract infections in spinal cord injury (SCI) patients. OBJECTIVES: To check whether the regular (1/week) urine cultures allow a more accurate treatment of urinary tract infections in SCI patients compared to empiric treatment. SETTING: Ghent University Hospital, East-Flanders, Belgium. METHODS: Group 1: 24 tetraplegic patients; group 2: 22 paraplegic patients; group 3: 28 other polytrauma patients as controls. These groups were chosen as catheterisation and other voiding methods differ according to the underlying pathology. RESULTS: An average of four clinically significant episodes of bacteriuria were found for groups 1 and 2, while group 3 experienced very few urinary infections. The mean species turnover of the first two groups was 2. No statistically significant difference was found in antibiotic-resistance patterns of organisms isolated. CONCLUSION: Despite different catheterisation techniques in para- and tetraplegic patients, we conclude that: (1) the number of episodes of clinical significant nosocomial urinary infections is not different; (2) the mean species turnover is the same; (3) because of the species turnover, the value of regular urine cultures for 'documented' treatment of clinical relevant urinary infections seems to be limited. So urine culture could be performed less frequently or only when therapy becomes mandatory; (4) No oral antibiotic with superior activity was found: treatment is best started empirically (after sampling for urine culture) and adjusted to the resulting antibiotic sensitivity screening.

Randomised controlled trial of nitrofurantoin versus placebo in the treatment of uncomplicated urinary tract infection in adult women.

BACKGROUND: Urinary tract infections (UTIs) are very common and have been treated with apparent success with antimicrobials for many years. However, there is a paucity of placebo-controlled clinical trials. AIM: To measure the symptomatic and bacteriological short-term effect of nitrofurantoin treatment versus placebo, in the treatment of uncomplicated UTI in adult non-pregnant women. DESIGN OF STUDY: Randomised placebo-controlled trial in general practice. SETTING: Non-pregnant women, aged between 15 and 54 years old, consulting a general practitioner for symtoms suggestive of uncomplicated lower UTI and with pyuria (positive for leucocyte esterase test). METHOD: A dipslide was inoculated in first-void midstream urine and sent for examinion. The patients were randomised to receive nitrofurantoin 100 mg or placebo four times daily for three days. After three, seven, and 14 days a new dipslide was inoculated and symptoms of UTI were checked or improvement of symptoms and bacteriuria. RESULTS: Of 166 women consulting with symptoms suggestive for UTI, 78 had pyuia and agreed to participate in the study (the clinically suspected UTI group); of these, 40 received nitrofurantoin and 38 received placebo. The result for combined symptomatic improvement and cure after three days was 27/35 in the nitrofurantoin group and 19/35 in the placebo group (c2 with Yates' correction P = 0.008; number needed to treat [NNT] = 4.4, 95% confidence interval [CI] = 2.3 to 79). After seven days, combined improvement and cure was observed in 30/34 and 17/33 respectively (P = 0.003, NNT = 2.7, 95% CI = 1.8 to 6.0). At inclusion, 56 women had bacteriuria of > or = 10(5) CFU/ml (the bacteriologically proven UTI group). Of these, 29 received nitrofurantoin and 27 received placebo. After three days the bacteriological cure was 21/26 in the treatment group, compared with 5/25 in the placebo group (P < 0.001; NNT = 1.6, 95% CI= 1.2 to 2.6). After seven days the bacteriological cure rate was 17/23 in the intervention group and 9/22 in the placebo group (P = 0.05, NNT = 3, 95% CI = 1.7 to 17). CONCLUSION: In women with bacteriologically proven UTI, nitrofurantoin was significantly more effective than placebo in achieving bacteriological cure and symptomatic relief in just three days; this was still present after seven days. In patients with clinically suspected UTI the symptomatic effect was statistically significant after

Identification of a new biotype of Actinomyces hyovaginalis in tissues of pigs during diagnostic bacteriological examination.

Bacterial strains isolated from a large variety of necropsy samples of pigs and previously described as a phenotypical homogeneous group were shown to belong to the species Actinomyces hyovaginalis. This was unexpected because their colonial characteristics, as well as their origins, were very different from those originally reported for the vaginal strains on which the species description of A. hyovaginalis was based. Colonial morphology, as well as fermentation of cellobiose, reactions in hippurate and nitrate and production of beta-glucuronidase, allowed separation of the strains studied here from the vaginal strains. Analysis of tRNA intergenic length polymorphisms (tDNA-PCR), 16S rRNA-gene sequencing and DNA-DNA hybridizations were carried out and led to the proposal of a separate biotype within the species A. hyovaginalis. Since, the strains were isolated from different body sites, this biotype has been designated as the 'general' biotype of A. hyovaginalis, while the strains on which the original species description was based are designated as the 'vaginal' biotype.

Case of catheter sepsis with Ralstonia gilardii in a child with acute lymphoblastic leukemia.

Acute lymphoblastic leukemia was diagnosed in a 7-year-old girl. Two months after insertion of a central venous catheter, she developed fever and complained of headache and abdominal pain. Physical examination revealed no focus of infection. A gram-negative nonfermenting bacillus was recurrently cultured from blood. Extensive biochemical testing and 16S ribosomal DNA sequencing led to the identification of Ralstonia gilardii.

One case each of recurrent meningitis and hemoperitoneum infection with Ralstonia mannitolilytica.

Two clinical cases of infection with Ralstonia mannitolilytica are described: a recurrent meningitis on an implanted intraventricular catheter and an infected hemoperitoneum as a complication of a cholangiocarcinoma. The strains were first misidentified as Pseudomonas fluorescens and Burkholderia cepacia. Further testing lead to the identification as Ralstonia pickettii biovar 3/"thomasii," which was recently shown to represent a separate species, R. mannitolilytica (List editor N. Weiss, Int. J. Syst. Evol. Microbiol. 51:795-796, 2001), originally described as R. mannitolytica (De Baere et al., Int. J. Syst. Evol. Microbiol. 51:547-558, 2001). R. mannitolilytica can be distinguished from all described Ralstonia species by its acidification of D-arabitol and mannitol and by its lack of nitrate reduction and of alkalinization of tartrate. In order to determine the true prevalence of infections with this species, colistin-resistant "P. fluorescens" strains and strains growing on B. cepacia selective medium deserve further attention.

Methicillin resistance of bacteria isolated from vitreous fluid from patients undergoing vitrectomy.

PURPOSE: The aim of this study was to compare the resistance patterns of bacteria in vitreous fluid from patients undergoing vitrectomy for diagnostic reasons, with bacteria of other nosocomial infections. METHODS: Vitreous fluid samples (n=144) were obtained from 133 patients undergoing vitrectomy for endophthalmitis, and 11 for uveitis as suspected endophthalmitis. They were Gram stained and cultured. Antibiotic susceptibility tests were run on all isolates. RESULTS: Gram stains were positive in 45/144 cases (31%), among which 38/45 (84%) were confirmed by a positive culture. Cultures were positive in 74/144 patients (51%) with mainly coagulase-negative staphylococci (n = 44) and Staphylococcus aureus (n = 13). In 133 patients endophthalmitis occurred after lens implantation (80 cases) and in 53 cases there was another origin (e.g. corneal transplantation, endogenous). In 26/80 post-lens implantation infections, culture remained negative; 32 infections occurred with coagulase-negative staphylococci, 10 with Staphylococcus aureus, 9 with streptococci and 3 with gram-negative bacteria. For endophthalmitis, ophthalmologists in our institution give an intraocular injection of vanccmycin and ceftazidim after vitrectomy. Among the 44 isolates of coagulase-negative staphylococci, 12 (27%) were resistant to methicillin. This is in contrast to other hospital-related coagulase-negative staphylococcus infections in general, and the resistance rate is 75% in our hospital. Only 2/13 Staphylococcus aureus isolates were methicillin-resistant. CONCLUSIONS: We conclude that isolates of coagulase-negative staphylococci from vitreous fluid are less resistant to methicillin than those isolated in other nosocomial infections.

[Avoidable "fever of unknown origin" due to Ralstonia pickettii bacteremia]. / Vermijdbare "febris e causa ignota" door Ralstonia pickettii bacteriemie.

Ralstonia pickettii bacteraemia from the atrial part of a ventriculo-atrial shunt was diagnosed as the cause of recurring febrile episodes with systemic toxicity in a 38-year old female. A prior neurosurgical resection of intracerebral cavernoma was complicated by postoperative recurrent meningitis with this non-fermenting gram-negative rod, due to an intra-operative contamination of a in situ shunt, from which only the intracerebral part of the ventriculo-atriostomy had been removed, following the episodes of meningitis. Ralstonia pickettii is a rare isolate and is seldom identified as a human pathogen. It formerly belonged to the genera Pseudomonas and Burkholderia. There are some casuistic reports of nosocomial bacteraemias, associated with the use of contaminated IV products or aerosols. There was an important diagnostic delay due to (1) insufficient communication between the different medical doctors in several hospitals, in different episodes of the clinical follow-up and (2) a misinterpretation of a positive blood culture with R. pickettii, which was interpreted as contamination. This misinterpretation and insufficient data transfer caused a considerable delay in the diagnostic process.

Modified Granada Agar Medium for the detection of group B Streptococcus carriage in pregnant women.

OBJECTIVES: To improve the detection rate of group B streptococci (GBS) in pregnant women, aiming at the prevention of early-onset septicemia in the newborn. METHODS: The yield from culturing two sites, vaginal and anorectal, on a Modified Granada Medium (MGM) was compared with our standard approach of culturing a vaginal swab on blood agar (BA). RESULTS: Samples were processed from 430 consecutive pregnant women. GBS was isolated from the vagina in 11.6% with BA, and in 13.7% with MGM. In 17.0% of anorectal samples, GBS was identified with MGM. The combination of both sites and media had a yield of 20.0%. MGM identified all but six (2%) of 310 GBS strains after aerobic incubation, with use of a cover slide, and missed only three strains (1%) after anaerobic incubation. CONCLUSIONS: Separate culture of vaginal and anorectal samples using the same MGM agar plate resulted in an increase in detection rate for GBS of 76% as compared to BA alone. The technique is simple and results are available after overnight incubation. MGM was confirmed as a specific medium for the identification of GBS, with a sensitivity of 98-99%.

Application and evaluation of the interlaboratory reproducibility of tRNA intergenic length polymorphism analysis (tDNA-PCR) for identification of Streptococcus species.

The discriminatory power, speed, and interlaboratory reproducibility of tRNA intergenic length polymorphism analysis (tDNA-PCR) combined with capillary electrophoresis was evaluated for the identification of streptococci. This method was carried out in three different laboratories under highly standardized conditions for 54 strains belonging to 18 different species. It was concluded that interlaboratory reproducibility of tDNA fingerprints produced by means of capillary electrophoresis was sufficiently high to permit the exchange between different laboratories and the construction of common libraries which can be consulted for comparison with fingerprints obtained independently in separate laboratories. In a second step, 17 other species were included in the study and examined in one of the participating laboratories. All Streptococcus species studied, except S. mitis, S. oralis, S. parasanguinis, S. pneumoniae, S. thermophilus, and S. vestibularis, showed distinguishable tDNA fingerprints. A database of well-characterized strains was constructed to enable computer-aided identification of unknown streptococcal isolates.

Isolations of Leclercia adecarboxylata from a patient with a chronically inflamed gallbladder and from a patient with sepsis without focus.

Leclercia adecarboxylata was isolated from a patient with a chronically inflamed gallbladder, together with Enterococcus sp. The organism was considered clinically significant and was susceptible to all antibiotics tested. Another strain of L. adecarboxylata was cultured from blood, together with Escherichia hermannii and E. faecalis, from a patient with sepsis.

Classification of Ralstonia pickettii biovar 3/'thomasii' strains (Pickett 1994) and of new isolates related to nosocomial recurrent meningitis as Ralstonia mannitolytica sp. nov.

Strains isolated independently from two patients could be recognized as Ralstonia pickettii biovar 3/'thomasii'. The 16S rDNA sequences of these strains and two other strains of R. pickettii biovar 3/'thomasii' clustered at less than 98% similarity versus all other described Ralstonia species and at less than 97 % versus the two other R. pickettii biovars. The separate species status of R. pickettii biovar 3/'thomasii' was confirmed by DNA-DNA hybridization, indicating less than 60% DNA homology with the R. pickettii biovars Va-1 and Va-2 and with two as-yet unclassified but biochemically similar Ralstonia strains. Phenotypically, this Ralstonia species can be distinguished from all described Ralstonia species by its acidification of D-arabitol and mannitol and by its lack of nitrate reduction and of alkalinization of tartrate and from two as-yet unclassified Ralstonia strains only by its lack of nitrate reduction. The name Ralstonia mannitolytica sp. nov. is proposed, reflecting the characteristic acidification of mannitol. Resistance to desferrioxamine is another difference from R. pickettii and Ralstonia solanacearum. Although several nosocomial outbreaks have been associated with R. mannitolytica, life-threatening infections have not yet been reported, possibly due to misidentification as Pseudomonas fluorescens or Burkholderia cepacia. In at least one of the two cases reported here, the R. mannitolytica isolate was found to be clinically relevant, causing recurrent nosocomial meningitis, with an infected implanted catheter as the source. The type strain of R. mannitolytica is NCIMB 10805T (= LMG 6866T), which was isolated during the first described outbreak as 'Pseudomonas thomasii' at St Thomas' Hospital, London, UK, in 1971.

Evaluation of six commercial assays for the rapid detection of Clostridium difficile toxin and/or antigen in stool specimens.

OBJECTIVE: To evaluate six commercially available assays for the detection of Clostridium difficile toxin and/or antigen in stool samples: one latex agglutination test (Culturette brand CDT, Becton Dickinson), two ELISAs (Culturette brand Toxin CD, Becton Dickinson, and Ridascreen C. difficile Toxin A/B, R-biopharm), two chromatographic assays (Clearview C. difficile A, Oxoid, and ColorPac Toxin A, Becton Dickinson) and one enzyme immunoassay for the simultaneous detection of C. difficile common antigen and toxin A (Triage C. difficile Panel, Biosite). METHODS: Over a period of 3 months, 366 liquid or semi-liquid stool samples were tested using cell-culture cytotoxin assay as standard, ethanol shock stool culture and latex agglutination (Culturette brand CDT). Of these, 78 samples, positive with at least one of these three methods, and 98 randomly selected negative samples were further evaluated using the other five kits. PCR was also performed on positive cultures to confirm the presence of toxin A and B genes. RESULTS: Triage C. difficile Panel had the best sensitivity (95%), followed by Clearview C. difficile and ColorPac Toxin A (both 89%), Culturette brand Toxin CD (73%), Ridascreen C. difficile Toxin A/B (57%) and Culturette brand CDT (23%). For Triage, the sensitivity of C. difficile antigen detection was 93%, and the sensitivity of toxin detection was lower (77%). Most false-positive results were obtained with the Triage C. difficile Panel (25 specimens) and Clearview C. difficile A (20 specimens). Culturette brand CDT had the best specificity (99%); followed by Ridascreen C. difficile Toxin A/B (97%), Culturette brand Toxin CD (95%), ColorPac Toxin A (89%), Clearview C. difficile A (83%) and Triage C. difficile Panel (75%). The positive predictive values ranged from 68% to 94%, and the negative predictive values from 83% to 98%. CONCLUSIONS: The sensitivity is much higher for Triage and the two new chromatographic assays than for the conventional EIAs. These tests also have a high negative predictive value. For Triage, C. difficile antigen-positive, toxin A-negative results can be obtained; the clinical value of these must be established by additional studies. Overall, the new-generation assays are still less sensitive than the cytotoxin assay; however, they provided same-day results, could be used as a screening test and may be useful in laboratories without tissue-culture facilities. Our results do not allow the recommendation of one single assay for the diagnosis of C. difficile-associated diarrhea. It remains the case that laboratory results must be correlated and interpreted with the clinical presentation of the patient.

Mycobacterium interjectum as causative agent of cervical lymphadenitis.

A mycobacterial strain isolated from a lymph node of a 3-year-old female with cervical lymphadenitis was identified as Mycobacterium interjectum by means of sequencing of the 16S rRNA gene. Analysis of this case and previously published cases demonstrates the importance of M. interjectum as a causative agent of cervical lymphadenitis in young children.

Application of tRNA intergenic spacer PCR for identification of Enterococcus species.

tRNA intergenic spacer PCR (tDNA-PCR) was evaluated for its usefulness in the differentiation of enterococcal species of human and animal origin. This technique was carried out for 124 strains belonging to 17 enterococcal species and generated DNA fragments, which were separated by capillary electrophoresis. tDNA-PCR enabled us to discriminate for all species tested. Enterococcus faecium showed minor but reproducible differences with Enterococcus durans, while Enterococcus hirae was easily distinguishable. Enterococcus avium, Enterococcus malodoratus, and Enterococcus raffinosus generated highly similar though distinctive patterns.