Bacteremic Isolates Research Articles

Virulence Factor Profiles and Phylogenetic Background ofEscherichia coliIsolates from Veterans with Bacteremia and Uninfected Control Subjects

Escherichia coli is the most common cause of gram-negative bloodstream infections, causing an estimated 40,000 deaths from sepsis each year in the United States. The present study sought to determine specifically which virulence factors (VFs) and phylogenetic groups of E. coli are epidemiologically associated with bacteremia. E. coli isolates from 63 veterans with bacteremia and rectal isolates from 71 matched uninfected control subjects were compared both for phylogenetic group and for the presence of VFs and O antigens. Bacteremia isolates exhibited a significantly greater prevalence of most VFs studied. In multivariate logistic regression analysis, ompT (outer membrane protein T) was the strongest VF predictor of bacteremia (P<.001). Despite the concentration of most individual VFs within group B2, bacteremia and rectal isolates differed little by phylogenetic distribution, a finding explained by the greater prevalence of VFs among bacteremia isolates than rectal isolates within groups B2 and D. Although phylogenetic group partially corresponds with virulence potential in E. coli bacteremia, VFs are more-powerful predictors of pathogenic potential. Bacteremia isolates exhibit an arsenal of VFs that distinguishes them from rectal isolates from uninfected hosts, which makes these differences attractive potential targets in vaccine or drug development.

Transcription of quorum-sensing system genes in clinical and environmental isolates of Pseudomonas aeruginosa.

Quorum sensing (QS)-based transcriptional responses in Pseudomonas aeruginosa have been defined on the basis of increases in transcript levels of QS-controlled genes such as lasB and aprA following the hierarchical transcriptional increases of central controllers such as the lasR gene. These increases occur at high bacterial concentrations such as early-stationary-phase growth in vitro. However, the extent to which the increases occur in a variety of clinical and environmental isolates has not been determined nor is there extensive information on allelic variation in lasR genes. An analysis of the sequences of the lasR gene among 66 clinical and environmental isolates showed that 81% have a sequence either identical to that of strain PAO1 or with a silent mutation, 15% have nucleotide changes resulting in amino acid changes, and 5% have an insertion sequence in the lasR gene. Using real-time PCR to quantify transcript levels of lasR, lasB, and aprA in the early log and early stationary phases among 35 isolates from bacteremia and pneumonia cases and the environment, we found most (33 of 35) strains had increases in lasR transcripts in early stationary phase but with a very wide range of final transcript levels per cell. There was a strong correlation (r(2) = 0.84) between early-log- and early-stationary-phase transcript levels in all strains, but this finding remained true only for the 50% of strains above the median level of lasR found in early log phase. There were significant (P < 0.05) but weak-to-modest correlations of lasR transcript levels with aprA (r(2) = 0.2) and lasB (r(2) = 0.5) transcript levels, but again this correlation occurred only in the 50% of P. aeruginosa strains with the highest levels of lasR transcripts in early stationary phase. There were no differences in distribution of lasR alleles among the bacteremia, pneumonia, or environmental isolates. Overall, only about 50% of P. aeruginosa strains from clinical and environmental sources show a lasR-dependent increase in the transcription of aprA and lasB genes, indicating that for about 50% of clinical isolates this regulatory system may not play a significant role in pathogenesis.

Ciprofloxacin-resistant Escherichia coli from bacteraemias in England; increasingly prevalent and mostly from men.

To assess ciprofloxacin resistance among Escherichia coli isolates from bacteraemia patients in England in relation to age, sex and Region. Routine susceptibility data for bacteraemia isolates were collected from over 90% of hospitals in England. During 1995-2001, the prevalence of ciprofloxacin resistance trebled, from 2.1% to 6.5%. Isolates from men were more frequently resistant than those from women, possibly because infections in men more often involve nosocomial strains. Resistance was rare (<1.5%) in isolates from patients aged <1 year; among older patients, resistance was unrelated to age in isolates from women, but peaked in the 15-44 age group for men. The prevalence of ciprofloxacin resistance in E. coli from bacteraemia is strongly associated with sex and, to a lesser extent, age.

Fatal outcome of bacteraemic patients caused by infection with staphylokinase-deficient Staphylococcus aureus strains.

Staphylokinase (SAK) is a plasminogen-activator protein produced by Staphylococcus aureus. SAK production was evaluated in vitro in S. aureus isolates from the bloodstream of patients with lethal (n = 56) and non-lethal (n = 57) bacteraemia and from anterior nares of healthy subjects (n = 48). Most isolates (93/161) produced SAK, and 68 % of SAK-producing isolates expressed both surface-bound and secreted types of SAK. SAK production was significantly less common among isolates from patients with lethal bacteraemia (39 %) than isolates from patients with non-lethal bacteraemia (68 %) or nasal carriage isolates (67 %) (P < 0.01). After adjusting for infection with methicillin-resistant S. aureus and APACHE II score, patients infected with SAK-deficient isolates were 4.3 times more likely to have lethal bacteraemia than patients whose infecting isolate produced high levels of SAK (> or =5 microg ml(-1)), suggesting that in vitro SAK production was inversely associated with clinical outcome among patients with S. aureus bacteraemia. The high frequency of SAK production in nasal isolates and in cases with uncomplicated bacteraemia suggests that SAK may be one of the adaptive mechanisms of S. aureus symbiosis with the host.

Klebsiella typing: pulsed-field gel electrophoresis (PFGE) in comparison with O:K-serotyping

To compare pulsed-field gel electrophoresis (PFGE) typing and O:K-serotyping of Klebsiella in two different epidemiological settings. One hundred and four bacteremia isolates without known epidemiological relation and 47 isolates from an outbreak in a neonatal intensive care unit (NICU) were K-typed by countercurrent immunoelectrophoresis (CCIE), O-typed by an inhibition enzyme-linked immunosorbent assay method, and typed by pulsed-field gel electrophoresis (PFGE) using the restriction enzyme XbaI. Typing data for the 104 bacteremia isolates were compared with regard to typability, number of types, maximum number of isolates per type, and the Discriminative Index (DI). O-typing combined with K-typing (DI 0.98) as O:K-serotyping (DI 0.99) gave a very discriminative typing system, whereas O-typing alone was not very discriminative (DI 0.76). PFGE (DI 1.00) was a more discriminative typing method than O:K-serotyping, as it could subdivide 13/22 O:K-serotypes into smaller groups. Isolates with the same PFGE-type had the same O:K-serotype, indicating that isolates with different O- and/or K-types could be expected to be of different PFGE-types. Typing of the 47 isolates from the outbreak in the NICU showed that 38 isolates belonged to a single clone, and that during an epidemic limited in time and space, differences in the electrophoretic patterns of up to five bands between a parental pattern type and a subtype may be found in the PFGE profiles. Both O:K-serotyping and PFGE typing are highly discriminative typing methods. PFGE is the most discriminative method and is excellent for typing outbreaks with few isolates. If large numbers of isolates are to be typed, a more convenient strategy might be first to K- or O:K-serotype isolates followed by PFGE typing of possible identical isolates. Since K- or O:K-serotyping is a definitive typing method, while PFGE typing is a comparative one, PFGE cannot, for the time being, replace O:K-serotyping for surveillance purposes.

Rapid method of extraction and analysis of extended-spectrum β-lactamases from clinical strains of Klebsiella pneumoniae

The extraction of periplasmic beta-lactamases from Gram-negative bacilli is a necessary preliminary step to analytical isoelectric focusing. Previously described methods are time-consuming and require large amounts of broth. We describe a lysozyme-based method which needs just 5 mL broth and requires less than 24 h to perform. The method was reproducible in extracting beta-lactamases from reference strains containing known beta-lactamases. We applied the method to a collection of more than 70 extended-spectrum beta-lactamase-producing isolates from a multinational study of bacteremic isolates of Klebsiella pneumoniae. Further studies are being undertaken to assess the method's applicability to other bacterial species.

Recurrent pneumococcal bacteremia: risk factors and outcomes.

Recurrent pneumococcal bacteremia receives infrequent mention in the literature, usually in association with patients who are immunocompromised. To examine recurrent cases of pneumococcal bacteremia to determine risk factors and outcomes (mortality rates and emergence of resistance) associated with recurrences. We retrospectively reviewed all cases of pneumococcal bacteremia identified by our microbiology laboratory from January 1, 1992, through December 31, 1996. Demographic, clinical, and laboratory data were abstracted. There were 462 bacteremic episodes in 432 patients; 23 of these patients had 30 recurrent episodes. The 5.3% recurrence rate (23/432) is greater than that previously described. The median time to recurrence was 200 days. The mean age of patients with recurrences was 34 years, 70% were women, all were black or Hispanic (in near equal numbers), and 87% were infected with the human immunodeficiency virus (HIV). Human immunodeficiency virus infection, coexistent cancer, and female sex were independent predictors of recurrence. Only patients who were HIV-infected had multiple recurrences. Isolates from recurrent bacteremias were more likely to be penicillin-resistant than were initial bacteremic isolates (relative risk, 2.0; P =.16). Patients with recurrences had a higher (although not statistically significant) mortality rate than those without recurrences (22% vs 16%; P =.33). There was an inverse relationship between severity of illness and likelihood of recurrence. Rates of recurrent pneumococcal bacteremia may be higher than previously reported. In patients with recurrent pneumococcal bacteremia, the presence of an underlying immunodeficiency should be investigated.

Molecular analysis of coagulase-negative Staphylococcus isolates from blood cultures: prevalence of genotypic variation and polyclonal bacteremia.

Fifty-seven coagulase-negative Staphylococcus isolates from 22 inpatients who had > or =2 blood cultures that were positive for Staphylococcus within 24 hours were analyzed to determine the frequency of polyclonal bacteremia. Patients were considered to have bacteremia (14 patients) or contamination of sample (8 patients) on the basis of clinical criteria. Nine colonies were randomly selected from each blood culture and genotyped by means of SmaI digestion/pulsed-field gel electrophoresis. Relatedness was determined by calculation of the Dice coefficient of banding-pattern similarity (S(AB)). Analysis of bacteremic isolates demonstrated the presence of a single species in 35 of 41 blood cultures, 1 related variant in 5 blood cultures (87%-92% S(AB)), and an unrelated strain in 1 blood culture (79% S(AB)). Analysis of contaminated samples demonstrated the presence of a single strain in 10 of 16 blood cultures and 1-3 variants (28%-97% S(AB)) in the remainder. Genotype diversity was significantly more common in the contaminated samples (P=.036). Almost all coagulase-negative Staphylococcus bacteremias were monoclonal.

Ongoing horizontal and vertical transmission of virulence genes and papA alleles among Escherichia coli blood isolates from patients with diverse-source bacteremia.

The phylogenetic distributions of multiple putative virulence factors (VFs) and papA (P fimbrial structural subunit) alleles among 182 Escherichia coli blood isolates from patients with diverse-source bacteremia were defined. Phylogenetic correspondence among these strains, the E. coli Reference (ECOR) collection, and other collections of extraintestinal pathogenic E. coli (ExPEC) was assessed. Although among the 182 bacteremia isolates phylogenetic group B2 predominated, exhibited the greatest concentration of individual VFs, and contained the largest number of familiar virulent clones, other phylogenetic groups exhibited greater concentrations of certain VFs than did group B2 and included several additional virulent clones. Certain of the newly detected VF genes, e.g., fyuA (yersiniabactin; 76%) and focG (F1C fimbriae; 25%), were as prevalent or more prevalent than their more familiar traditional counterparts, e.g., iut (aerobactin; 57%) and sfaS (S fimbriae; 14%), thus possibly offering additional useful targets for preventive interventions. Considerable diversity of VF profiles was observed at every level within the phylogenetic tree, including even within individual lineages. This suggested that many different pathways can lead to extraintestinal virulence in E. coli and that the evolution of ExPEC, which involves extensive horizontal transmission of VFs and continuous remodeling of pathogenicity-associated islands, is a highly active, ongoing process.

Antibiotic sensitivity still prevails in Norwegian blood culture isolates

We describe the antimicrobial susceptibility of bacteraemia isolates from Norway. From March 1998 to February 1999, four university hospitals covering all parts of Norway collected their first 10 isolates each month. Minimal inhibitory concentrations were determined for: Enterobacteriaceae ( n=192), staphylococci ( n=89) and Streptococcus pneumoniae ( n=69) using the Etest. NCCLS breakpoints were used. About 20% of all blood culture isolates in Norway in this period were investigated. Compared with countries outside Scandinavia antibiotic sensitivity still prevails. Only minor differences in resistance were found between participating hospitals, between hospital departments and between hospital- and community-acquired pathogens. The prudent use of antibiotics in Norway may contribute to the fact that antibiotic resistance still remains low in the most common bacterial pathogens causing bloodstream infections.

Molecular investigation of two clusters ofhospital-acquired bacteraemia caused by multi-resistant Klebsiella pneumoniae using pulsed-field gel electrophoresis andinfrequent restriction site PCR

Two molecular typing methods, DNA macrorestriction analysis with XbaI resolved by pulsed-field gel electrophoresis (PFGE) and infrequent restriction site PCR (IRS-PCR) assay with adapters designed for XbaI andHhaI restriction sites, were used to investigate two clusters of hospital-acquired bacteraemia associated with multi-resistant Klebsiella pneumoniae which occurred in a paediatric intensive care unit (PICU). A total of 56 K. pneumoniae isolates were analysed. These included 10 bacteraemic isolates from eight patients, 26 isolates obtained during an epidemiological survey, and 20 epidemiologically non-related isolates incorporated as controls. One major pattern was demonstrated in 22 of the 56 isolates analysed. These included nine of the 10 bacteraemic isolates, a single rectal isolate, two hand culture isolates and 10 sink isolates. All of these 22 isolates illustrated identical antibiograms, whilst the other 34 isolates shared six antibiograms and 31 unique patterns by either PFGE or IRS-PCR assay. The two clusters of bacteraemia appeared to be outbreaks induced by the same strain of K. pneumoniae which may have utilized sinks as reservoirs and been transmitted through the hands of medical personnel to patients. IRS-PCR demonstrates concordant results with PFGE analysis in studying the genetic relationships among K. pneumoniae isolates, and serves as an excellent epidemiological tool for this bacterium.

Regional variation in ampicillin and trimethoprim resistance in Escherichia coli in England from 1990 to 1997, in relation to antibacterial prescribing.

Over 200 hospitals in England report resistance data for bacteraemia and meningitis isolates to the Public Health Laboratory Service. We reviewed ampicillin and trimethoprim resistance rates from 1990 to 1997 for Escherichia coli, which is the species reported most frequently from these bacteraemias. Ampicillin resistance was relatively stable over time, but varied between Health Regions. The proportion of ampicillin-resistant E. coli in the East Anglia region remained </=42% in all years except one and that in the South Western region always remained <50%. At the other extreme, the proportions of ampicillin-resistant isolates in the Northern and Trent regions never fell below 59%. The prevalence of resistance to trimethoprim rose over time in most regions; again, however, the prevalence of resistant isolates was lowest in the East Anglia and South Western regions, whereas the highest resistance rates were reported from Mersey, NW Thames, NE Thames and North Western regions. These observations were related to data for community prescribing, which accounts for most ampicillin and trimethoprim use. Prescribing data for ampicillin and trimethoprim from 1987 to 1997 were obtained from the IMS-HEALTH Medical Data Index, and data for all antibacterial drugs between 1995 and 1997 from the Prescription Pricing Authority. Correlations between resistance rates and prescribing of specific antibiotics were weak, although there was some trend for regions with high total prescribing to have higher rates of ampicillin resistance. The South Western region was conspicuous both for low rates of resistance and low prescribing. Several factors may determine the lack of wider and more obvious relationships between resistance and prescribing. In particular, regions may be inappropriately large areas to test the relationship, isolates from bacteraemias may not be representative of those experiencing selection pressure in the community and the resistance data may have been distorted by nosocomial strains, although this seems unlikely with E. coli.

Concordance of endotoxemia with gram-negative bacteremia. A meta-analysis using receiver operating characteristic curves.

OBJECTIVE: To apply meta-analysis to compare the concordance between the results of 2 types of limulus amebocyte lysate (LAL) assay, gelation (GLAL) and chromogenic (CLAL), with the detection of gram-negative bacteremia in patients with suspected bacteremia. DESIGN: Meta-analysis using receiver operating characteristic-based analytical method. DATA SOURCES: MEDLINE literature search and manual reviews of article bibliographies together with direct approaches to authors of potentially eligible studies. STUDY SELECTION: The studies that were selected had all included at least 10 patients, of whom at least 2 patients were diagnosed with gram-negative bacteremia, and all had data available for extraction into a contingency table format. RESULTS: Fifty-six studies (28 GLAL and 28 CLAL studies) met the inclusion criteria. Studies were stratified by type of test (GLAL vs CLAL). Each analysis was repeated with smaller studies excluded. There was no difference between the 2 types of LAL assays. Among the CLAL studies, there was no difference between studies that did versus those that did not use the sepsis syndrome criteria as a basis for patient inclusion. Among 45 studies for which data on the proportion of non-Enterobacteriaceae were available, there was a trend toward higher concordance as this proportion increased. CONCLUSIONS: The concordance between the LAL test and the detection of gram-negative bacteremia in patients with suspected bacteremia is no higher with the CLAL assay than with the original GLAL version. However, the concordance is higher among studies with a higher proportion of non-Enterobacteriaceae among the gram-negative bacteremia isolates.

A 5-year study of the seroepidemiology of Klebsiella pneumoniae: high prevalence of capsular serotype K1 in Taiwan and implication for vaccine efficacy.

Seroepidemiology of Klebsiella pneumoniae was determined for 1000 nonrepetitive K. pneumoniae isolates collected by a medical center in Taiwan during 1993-1997. Of these, 630 isolates (63%) were from community-acquired infections; the rest were from hospital-acquired infections. The isolates were serotyped according to capsular antigen by countercurrent immunoelectrophoresis. About 77% were typeable. Serotypes K1 and K2 accounted for 21.7% and 9.3% of the isolates, respectively, followed by K57 (5.1%), K54 (4.2%), K21 (3. 3%), and K16 (3%). The frequency of serotype K1 among bacteremic isolates (30.8%) far exceeded that reported by other investigators worldwide. Molecular typing of random K1 isolates by pulsed-field gel electrophoresis revealed several different pulsotypes, suggesting a nonclonal spread. This study indicates that a Klebsiella vaccine developed in Europe is not optimal for use in Taiwan because it does not contain the most predominant serotypes-K1, K54, and K57.

Outbreak investigation of nosocomial enterobacter cloacae bacteraemia in a neonatal intensive care unit.

Over a period of 7 months, 23 patients hospitalized in a neonatal intensive care unit (NICU) developed nosocomial Enterobacter cloacae bacteraemia. Contaminated saline for preparing heparin solution was initially identified as the common source of E. cloacae bacteraemia. Although environmental sanitation was enforced, the outbreak continued. E. cloacae has always been isolated from various cultures of the environmental specimens, from the hands of personnel and from the faeces of patients. All of the 23 bacteraemic isolates and 8 stool isolates from infected infants, as well as the 17 isolates from environmental specimens were found to be of the same genotype using the polymerase chain reaction-based DNA fingerprinting method. After various infection control methods were instituted, the outbreak eventually came under control. For epidemiological investigation, 23 neonates without E. cloacae bacteraemia were matched for case-control study. Nineteen (83%) of the case-patients were premature. The significant risk factors leading to E. cloacae bacteraemia in the NICU included small gestation age, low birthweight, exposure to personnel with contaminated hands and the presence of E. cloacae in the stool carriage (p=0.003, 0.007, 0.018 and 0.040, respectively). The gastrointestinal tracts of the patients and environmental surfaces appeared to be the principal sites of bacterial reservoir. In conclusion, the outbreak of E. cloacae bacteraemia was caused by a particular strain and possibly via multiple modes of transmission, including a bottle of contaminated saline as an initial common source, endogenous spread from the gastrointestinal tract and successive cross-infections between patients, hands of personnel and the environment. Effective infection control requires a multidisciplinary approach and reinforcement of infection control procedures, including aseptic technique, hand washing, proper isolation and disinfection of environmental surfaces.

Changing epidemiology of infections in patients with neutropenia and cancer: emphasis on gram-positive and resistant bacteria.

Over the past 3 decades, considerable changes have occurred in the types of bacteria causing infection in febrile patients with neutropenia and cancer. Twenty years ago, gram-negative bacteria caused approximately 70% of bloodstream infections. As a probable consequence of long-dwelling intravascular devices, fluoroquinolone prophylaxis, and high-dose chemotherapy-induced mucositis, there has been a shift toward gram-positive coccal bacteremia. In most centers today, approximately 70% of bacteremic isolates are gram-positive cocci. Of potential concern is that antimicrobial-resistant gram-positive organisms are becoming increasingly frequent in patients with neutropenia. Fluoroquinolone-resistant Escherichia coli are being isolated from several cancer centers. Several "new" organisms, such as Stomatococcus mucilaginosus, Bacillus cereus, Leuconostoc species, Corynebacterium jeikeium, Rhodococcus species, Stenotrophomonas maltophilia, Moraxella catarrhalis, Burkholderia cepacia, and Bartonella species, now cause infections in these patients. Careful application of infection-control principles, judicious prophylaxis, appropriate evaluation of new antibiotics, and prompt effective therapy will maximize benefits for these patients.

All Wales surveillance of methicillin-resistant Staphylococcus aureus (MRSA): The first year's results.

Over the last five years, hospitals in Wales have experienced difficulties with increasing numbers of isolates of methicillin-resistant Staphylococcus aureus (MRSA). Continuous total population surveillance of MRSA was introduced with the objectives of gaining an understanding of the extent and variation in time and place of its occurrence, the burden of disease and possible risk factors associated with its isolation and resistance to other antibiotics. All first isolates of MRSA from both hospital and community settings and all isolates of methicillin-sensitive Staphylococcus aureus (MSSA) associated with bacteraemia and cerebrospinal fluid (CSF) isolates detected in medical microbiology laboratories in Wales were collected via CoSurv, a set of interconnected database modules for communicable disease control. A data set was collected on each isolate and the patient associated with that isolate and compiled centrally at CDSC (Wales) for all-Wales analysis of the MRSA situation. Surveillance started in January 1996 and at the end of the first year, 2700 new isolates of MRSA had been reported from hospital and community settings, giving a rate of 92.43/100 000 population. The incidence of MRSA from bacteraemias and CSF was 5.20/100 000 compared with 12.70/100 000 for MSSA. MRSA from bacteraemia and CSF was significantly more commonly associated with male patients than MSSA. MRSA patients were significantly older. For all MRSA isolates, the highest reporting rate was in men aged 75+ (647.21/100 000). The highest incidence of invasive disease was also in men aged 75+ (45.69/100 000). Isolates from post-surgical patients were more likely to be involved in invasive disease (OR=2.59, P<0.001) than strains from other sources. The majority of isolates were resistant to at least two antibiotics in addition to methicillin, most frequently erythromycin and the fluoroquinolones. Very little resistance to fusidic acid, mupirocin or rifampicin was reported. Continuous total population surveillance has provided a minimum incidence of MRSA in Wales and has allowed a simple and intelligible picture of the problem to be determined, which has been fed back to hospitals to assist decisions on control

In vitro resistance to thrombin-induced platelet microbicidal protein among clinical bacteremic isolates of Staphylococcus aureus correlates with an endovascular infectious source.

Platelet microbicidal proteins (PMPs), small cationic peptides released at sites of endovascular damage, kill common bloodstream pathogens in vitro. Our group previously showed that in vitro resistance of clinical staphylococcal and viridans group streptococcal bacteremic strains to PMPs correlated with the diagnosis of infective endocarditis (IE) (Wu et al., Antimicrob. Agents Chemother. 38:729-732, 1994). However, that study was limited by (i) the small number of Staphylococcus aureus isolates from IE patients, (ii) the retrospective nature of the case definitions, and (iii) the diverse geographic sources of strains. The present study evaluated the in vitro PMP susceptibility phenotype of a large number of staphylococcemic isolates (n = 60), collected at a single medical center and categorized by defined and validated clinical criteria. A significantly higher proportion of staphylococcemic strains from patients with IE was PMP resistant in vitro than the proportion of strains from patients with soft tissue sepsis (83% and 33%, respectively; P < 0.01). Moreover, the levels of PMP resistance (mean percent survival of strains after 2-h exposure to PMP in vitro) were significantly higher for isolates from patients with IE and with vascular catheter sepsis than for strains from patients with abscess sepsis (P < 0.005 and P < 0.01, respectively). These data further support the concept that bloodstream pathogens that exhibit innate or acquired PMP resistance have a survival advantage with respect to either the induction or progression of endovascular infections.

Clonal Distribution of the Three Alleles of the Gal(α1–4)Gal‐Specific Adhesin GenepapGamongEscherichia coliStrains from Patients with Bacteremia

The three alleles of papG, the Gal(alpha1-4)Gal-specific adhesin gene of Escherichia coli P fimbriae, were detected in 101 (54%) of 187 E. coli bacteremia isolates from Boston, Long Beach, California, and Nairobi, Kenya, by a polymerase chain reaction-based assay, and their distribution was compared with clonal structure and other bacterial and host characteristics. Allele II predominated overall (57% of papG+ strains, vs. 32% for III and 2% for I). Allele distribution differed significantly between centers. Alleles segregated according to enzyme electrophoretic lineage and O serogroup in patterns suggesting both vertical and horizontal transmission and emergence within E. coli in the temporal sequence II-->III-->I. Strains containing allele III preferentially agglutinated sheep erythrocytes, whereas strains containing allele II preferentially agglutinated human erythrocytes. Allele III was associated with hly, sfa, multiple copies of the pap operon, and cluster III of the phylogenetic tree. These findings indicate that among bacteremia isolates of E. coli, the three papG alleles exhibit distinctive associations with evolutionary lineages, hemagglutination phenotypes, and non-pap virulence properties.

Epidemiological investigation of Pseudomonas aeruginosa nosocomial bacteraemia isolates by PCR-based DNA fingerprinting analysis.

Between July 1994 and March 1995, 64 isolates of Pseudomonas aeruginosa were implicated in bacteraemia in 25 cancer patients in five wards of two hospitals. These, together with 24 environmental isolates and one isolate from a bacteraemia in a non-cancer patient were examined by three PCR-based DNA fingerprinting methods: random amplified polymorphic DNA (RAPD), enterobacterial-repetitive intergenic consensus (ERIC)-PCR, and 16S-23S spacer region-based RAPD. These methods were reproducible, discriminatory and showed close agreement; all indicated that 47 isolates that had caused bacteraemia in 19 cancer patients were indistinguishable. Seventeen other isolates that had caused bacteraemia in 10 cancer patients were discriminated into eight further groups, and the 24 environmental and non-cancer patient isolates into further distinct groups. No environmental source of the epidemic strain was found, but it was suspected that the outbreak was related to infusion implants.