Outbreak of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae infection among renal transplant recipients.
Explore the source record for details and available documents.
Brazil↗
Biomedical subjects
L W Riley
Publications and source records attributed to L W Riley.
Survival characteristics of Salmonella enterica serovar Enteritidis in chicken egg albumen.
Salmonella enterica serovar Enteritidis (SE) is a major foodborne pathogen primarily causing human infection through contaminated chicken eggs. To understand how SE survives in chicken egg albumen, we systematically and quantitatively analysed the survival properties of SE in egg albumen and identified factors affecting its survival. Survival assays of SE in egg indicate that egg albumen restricted the growth of SE. A major factor that controlled SE's growth in egg albumen was iron restriction, since egg albumen supplemented with iron allowed SE to grow, and iron acquisition mutants of SE showed decreased survival in egg albumen. In addition, low pH of albumen, high concentrations of bacteria and low incubation temperatures of bacteria with albumen facilitates the survival of SE. Our results suggest that egg albumen uses multiple mechanisms to control SE including iron limitation, surface interaction and possible enzymatic activities.
Animals↗
Recent transmission of tuberculosis involving retired patients.
The reported incidence of tuberculosis (TB) in three different regions of Rio Grande do Sul State in Brazil varies considerably. We used IS6110-RFLP and spoligotyping methods to genotype Mycobacterium tuberculosis isolates obtained from 268 patients between 1998 and 2000 in order to assess the levels of recent transmission of TB in the three regions. The degree of clustering of the strain types did not differ among the three regions; neither did other characteristics such as demographic features, underlying medical conditions, or the proportion of resistant TB. As reported previously, male patients were at greater risk of developing TB and our data suggest that part of this may be related to the higher rates of recent transmission among them (P<0.05). In addition, we found that retired patients were almost 3 times more likely to be infected with cluster-pattern strains than patients reporting any other occupation (P<0.05), and more than 3 times more likely than non-retired patients in the same age group (P<0.05) to be infected with cluster-pattern strains. We conclude that recent transmission is not a major factor contributing to the differences in TB incidence in the three regions of Rio Grande do Sul. The reason for the suggested high proportion of recent transmission TB cases among the retired people needs further studies.
Adolescent↗
The changing prevalence of drug-resistant Escherichia coli clonal groups in a community: evidence for community outbreaks of urinary tract infections.
A multidrug-resistant clonal group (CgA) of Escherichia coli was shown to cause half of all trimethoprim-sulphamethoxazole (TMP-SMZ)-resistant urinary tract infections (UTIs) in a college community between October 1999 and January 2000. This second study was conducted to determine the fate of CgA. Urine E. coli isolates from women with UTI, collected between October 2000 and January 2001, were tested for antibiotic susceptibility, O serogroup, ERIC2 PCR and DNA macrorestriction patterns using pulsed-field gel electrophoresis. The proportion of UTIs caused by CgA declined by 38% (P<0.001) but the prevalence of resistance to TMP-SMZ did not change. Six additional clonal groups were identified and these were responsible for 32% of TMP-SMZ-resistant UTIs. The temporal decline in the proportion of UTIs caused by CgA provides evidence that CgA caused a community outbreak of UTI. The fluctuation and occurrence of other E. coli clonal groups in this community suggest that a proportion of community-acquired UTIs may be caused by E. coli disseminated from one or more point sources.
Adolescent↗
Pseudomonas aeruginosa epidemic strain carrying bla(SPM) metallo-beta-lactamase detected in Rio de Janeiro, Brazil.
The present study was designed to characterize beta-lactamase genes and evaluate polymerase chain reaction (PCR) typing for multidrug-resistant Pseudomonas aeruginosa pulsed-field gel electrophoresis (PFGE) genotype A isolates from Rio de Janeiro, Brazil, collected between April 1999 and March 2000 and one additional isolate collected in June 2002. As reported previously, all of the genotype A isolates produced non-characterized metallo-beta-lactamase. These isolates (22) were screened for the bla(SPM) gene by PCR and dot-blotting. Isolates were typed by PCR fingerprinting with primers RAPD-1, 272, 208, 1290, ERIC-1 and ERIC-2. The bla(SPM) gene was detected in 18 (82%) of the 22 isolates. PCR fingerprinting gave results that correlated with PFGE, except with primer 1290. In Rio de Janeiro and other Brazilian states, nearly all SPM-producing P. aeruginosa isolates belong to a single PFGE type accounting for a large proportion of drug-resistant P. aeruginosa hospital infections. RAPD PCR fingerprinting may be a useful technique to screen for an epidemic multidrug-resistant strain in Brazil.
Anti-Bacterial Agents↗
A population-based study of drug resistance and transmission of tuberculosis in an urban community.
SETTING: A low-income neighborhood of Sao Paulo, Brazil. OBJECTIVE: To determine the incidence, risk factors and transmission patterns of multidrug-resistant tuberculosis (MDR-TB). DESIGN: Prospective longitudinal study of patients with pulmonary TB (PTB). METHODS: Sputum culture-confirmed patients with PTB were recruited between March 2000 and May 2002. Bivariate and multivariate logistic regression analyses were performed to identify factors associated with MDR-TB. Mycobacterium tuberculosis isolates were tested for drug susceptibility and typed by IS6110-RFLP analysis. RESULTS: Of 420 patients, respectively 71% and 27% were new and previously treated; 15.5% of the patients' M. tuberculosis isolates were resistant to at least one drug; of these, 11% and 27% were found among new and previously treated cases, respectively. Respectively 1% and 16.7% of the new and previously treated cases were MDR-TB. RFLP analysis showed that new transmission of MDR strains was uncommon. By multivariate logistic regression analysis, previous TB and hospitalization in the 24 months before TB diagnosis were identified as independent predictors of MDR-TB. CONCLUSIONS: The results showed an intermediate level of MDR-TB incidence in a neighborhood of Sao Paulo and identified predictors that can be targeted for intervention by national and local TB control programs.
Adolescent↗
Integrated portable genetic analysis microsystem for pathogen/infectious disease detection.
An integrated portable genetic analysis microsystem including PCR amplification and capillary electrophoretic (CE) analysis coupled with a compact instrument for electrical control and laser-excited fluorescence detection has been developed. The microdevice contains microfabricated heaters, temperature sensors, and membrane valves to provide controlled sample positioning and immobilization in 200-nL PCR chambers. The instrument incorporates a solid-state laser and confocal fluorescence detection optics, electronics for sensing and powering the PCR reactor, and high-voltage power supplies for conducting CE separations. The fluorescein-labeled PCR products are amplified and electrophoretically analyzed in a gel-filled microchannel in <10 min. We demonstrate the utility of this instrument by performing pathogen detection and genotyping directly from whole Escherichia coli and Staphylococcus aureus cells. The E. coli detection assay consists of a triplex PCR amplification targeting genes that encode 16S ribosomal RNA, the fliC flagellar antigen, and the sltI shigatoxin. Serial dilution demonstrates a limit of detection of 2-3 bacterial cells. The S. aureus assay uses a femA marker to identify cells as S. aureus and a mecA marker to probe for methicillin resistance. This integrated portable genomic analysis microsystem demonstrates the feasibility of performing rapid high-quality detection of pathogens and their antimicrobial drug resistance.
Animals↗
Multiclonal Leishmania braziliensis population structure and its clinical implication in a region of endemicity for American tegumentary leishmaniasis.
In Corte de Pedra (CP), northeastern Brazil, Leishmania braziliensis causes three distinct forms of American tegumentary leishmaniasis (ATL). To test the hypothesis that strain polymorphism may be involved in this disease spectrum and accurately characterize the parasite population structure in CP, we compared one L. major, two non-CP L. braziliensis, one CP L. amazonensis, and 45 CP L. braziliensis isolates, obtained over a 10-year period from localized cutaneous, mucosal, and disseminated leishmaniasis patients, with randomly amplified polymorphic DNA (RAPD). Electrophoretic profiles were mostly unique across species. All typing protocols revealed polymorphism among the 45 CP L. braziliensis isolates, which displayed eight different RAPD patterns and greater than 80% overall fingerprint identity, attesting to the adequacy of the tools to assess strain variability in CP's geographically limited population of parasites. The dendrogram based on the sum of RAPD profiles of each isolate unveiled nine discrete typing units clustered into five clades. Global positioning showed extensive overlap of these clades in CP, precluding geographic sequestration as the mechanism of the observed structuralization. Finally, all forms of ATL presented a statistically significant difference in their frequencies among the clades, suggesting that L. braziliensis genotypes may be accompanied by specific disease manifestation after infection.
Animals↗
[Distribution of HBV genotypes in the Czech population--pilot study].
A compact structure of the hepatitis B virus genome does not provide much space for development of mutations capable of further replication. In spite of that, 7 genotypes have been described so far. Discovery of the hepatitis B virus (HBV) genome diversity led to a range of further questions. Individual genotypes are subjects of investigation these days as for the pattern of disease transmission, course of disease, and consequences of an infection. The aim of this pilot study was to describe genotype distribution in the infected population of the Czech Republic. Moreover, epidemiological and clinical characteristics were monitored in our sample. We studied a group of 65 patients with positive HBsAg from which 48 samples of patients were genotyped. PCR (polymerase chain reaction) method was used in order to amplify S region of the HBV genome. Direct sequencing of PCR products was used, this is the most accurate method for genotype identification. A relatively uniform genotypic distribution was identified within the study population--only 2 genotypes, A and D, were found. Genotype A was identified in 35 (73%) patients and genotype D in 13 (27%) patients. No statistically significant association between the particular genotype and epidemiological and clinical parameters was proved in this pilot study. This study brings basic information on genotype distribution. A possible proof of correlation with clinical and epidemiological characteristics will require examination of a larger sample of patients.
Adult↗
Extended-spectrum beta-lactamase-producing Klebsiella pneumoniae in a neonatal intensive care unit: risk factors for infection and colonization.
An outbreak of extended spectrum beta-lactamase-producing Klebsiella pneumoniae (ESBLKp) infections in a neonatal intensive care unit (NICU) prompted a prospective investigation of colonization and infection with this pathogen. From August 1, 1997 to May 30, 1999, neonates admitted to the NICU for more than 24 h were screened for ESBLKp acquisition. Neonatal gastrointestinal screening was performed by means of faecal sampling within 48 h of admission and then weekly until discharge. Isolates were typed using pulsed-field gel electrophoresis (PFGE). Time-dependent proportional hazard models were used to identify independent effects of invasive procedures and antimicrobials after controlling for duration of stay at the NICU. During the study period, 464 neonates were admitted and 383 were regularly screened. Infections occurred in 13 (3.4%) neonates and 206 (53.8%) became colonized. Independent risk factors for colonization during the first nine days in the NICU were the antimicrobial combination cephalosporin plus aminoglycoside [hazard rate (HR)=4.60; 95% CI: 1.48-14.31], and each NICU-day was associated with a 26% increase in the hazard rate for colonization (HR=1.26; 95% CI: 1.16-1.37). Previous colonization (HR=5.19; 95% CI: 1.58-17.08) and central vascular catheter use (HR=13.89; 95% CI: 2.71-71.3) were independent risk factors for infection. In an outbreak setting the proportion of neonates colonized with ESBLKp was observed to increase with the duration of stay and antimicrobial use, and once colonized, infants exposed to invasive devices may become infected.
Anti-Bacterial Agents↗
Energy transfer between fluorescent proteins using a co-expression system in Mycobacterium smegmatis.
The goal of this study was to establish a two-plasmid co-expression system for Mycobacterium smegmatis. Two vectors with compatible origins of replication and a polylinker, which allows modular cloning of promoters and genes, were constructed and used to clone genes encoding a blue fluorescent protein (BFP) and a green fluorescent protein (GFP). A 160-fold variation of GFP expression levels in M. smegmatis was achieved by combining three promoters with different copy numbers of the vectors. An efficient energy transfer between BFP and GFP in M. smegmatis was observed by fluorescence measurements and demonstrated that these genes were simultaneously expressed from both vectors. Thus, these vectors will be valuable for all strategies where co-expression of proteins in M. smegmatis is needed, e.g. for constructing a two-hybrid system or for deleting essential genes.
Base Sequence↗
Widespread distribution of urinary tract infections caused by a multidrug-resistant Escherichia coli clonal group.
BACKGROUND: The management of urinary tract infections is complicated by the increasing prevalence of antibiotic-resistant strains of Escherichia coli. We studied the clonal composition of E. coli isolates that were resistant to trimethoprim-sulfamethoxazole from women with community-acquired urinary tract infections. METHODS: Prospectively collected E. coli isolates from women with urinary tract infections in a university community in California were evaluated for antibiotic susceptibility, O:H serotype, DNA fingerprinting, pulsed-field gel electrophoretic pattern, and virulence factors. The prevalence and characteristics of an antibiotic-resistant clone were evaluated in this group of isolates and in those from comparison cohorts in Michigan and Minnesota. RESULTS: Fifty-five of the 255 E. coli isolates (22 percent) from the California cohort were resistant to trimethoprim-sulfamethoxazole as well as other antibiotics. There was a common pattern of DNA fingerprinting, suggesting that the isolates belonged to the same clonal group (clonal group A), in 28 of 55 isolates with trimethoprim-sulfamethoxazole resistance (51 percent) and in 2 of 50 randomly selected isolates that were susceptible to trimethoprim-sulfamethoxazole (4 percent, P<0.001). In addition, 11 of 29 resistant isolates (38 percent) from the Michigan cohort and 7 of 18 (39 percent) from the Minnesota cohort belonged to clonal group A. Most of the clonal group A isolates were serotype O11:H(nt) or O77:H(nt), with similar patterns of virulence factors, antibiotic susceptibility, and electrophoretic features. CONCLUSIONS: In three geographically diverse communities, a single clonal group accounted for nearly half of community-acquired urinary tract infections in women that were caused by E. coli strains with resistance to trimethoprim-sulfamethoxazole. The widespread distribution and high prevalence of E. coli clonal group A has major public health implications.
Adolescent↗
Genotypic and epidemiologic characteristics of hepatitis C virus infections among recent injection drug user and nonuser populations.
The recent surge in injection drug use (IDU) in Prague, Czech Republic, provided an opportunity to compare the distribution of hepatitis C virus (HCV) genotypes among recent injection drug users and nonusers. We provide evidence that the most common HCV subtype, 1b, has been circulating in Prague longer than the other subtypes but that IDU practice has magnified its recent prevalence, and that the primary drug of choice among IDU subjects appears to influence the prevalence of a particular HCV subtype.
Adult↗
Cost of hospitalization for and risk factors associated with vancomycin-resistant Enterococcus faecium infection and colonization.
The increase in costs of hospitalization for patients with drug-resistant infection may be associated with drug resistance itself or with the severity of the underlying illness that predisposes patients to acquire the drug-resistant infection. To address this issue, risk factors and cost of hospitalization were compared for patients infected or colonized with vancomycin-susceptible Enterococcus faecium (VSEF) or vancomycin-resistant E. faecium (VREF) in a large tertiary-care hospital in New York City. From January 1995 through December 1996, 157 patients with VSEF and 262 patients with VREF were identified. CMI (case-mix index) was assigned to each patient as a measure of severity of illness, with a CMI of 1 considered to represent illness of average severity. For all patients who were assigned a CMI of <3, the cost per day of hospitalization for patients with VREF was significantly greater than that for patients with VSEF. However, for patients with a CMI of >3, there was no difference between cost of hospitalization for patients with VREF and that for patients with VSEF. These observations indicate that, although vancomycin resistance is associated with an increased cost of hospitalization for less severely ill patients with VREF, patients with severe underlying illness, regardless of vancomycin resistance, incur similar hospitalization costs.
Adolescent↗
Molecular epidemiology of tuberculosis among eight hospitals in New York City, 1996-1997.
OBJECTIVE: To determine the molecular epidemiology of tuberculosis isolated from patients cared for at eight hospitals scattered throughout New York City. MATERIALS AND METHODS: Cases of tuberculosis occurring in 1996 and 1997 at collaborating hospitals were identified, and demographic data were extracted from patient charts. All available isolates were analyzed by IS6110 for genetic relatedness. The molecular fingerprints were compared both to each other and to the larger repository of strains from New York City developed and maintained at the Public Health Research Institute. RESULTS: One hundred and eighty cases were fully characterized. Compared with New York City cases, study patients were more likely to be Asian and less likely to be non-Hispanic blacks. Overall, 97 (54%) of the cases were clustered with respect to other study strains or with respect to the other New York City isolates. Clustered strains were significantly more likely to be from non-Hispanic blacks or patients born in the United States. The largest cluster (n = 17) was the "W" strain previously associated with an outbreak of multidrug-resistant tuberculosis in New York City. In the current study, the majority of W strain isolates were fully drug-susceptible. CONCLUSIONS: High rates of genetically related tuberculosis continue to occur among patients in New York City, in spite of improved control of nosocomial outbreaks and dramatic decreases in the overall case rates. The use of molecular techniques to suggest patterns of transmission has become essential in developing and assessing routine tuberculosis control strategies.
Adult↗
Genotypic distribution of TT virus (TTV) in a Czech population: evidence for sexual transmission of the virus.
BACKGROUND: TTV is a new DNA virus distinguished by its high degree of strain heterogeneity. The geographic clustering of viral genotypes suggests frequent community transmission. While no specific human disease has yet been linked to it, a transmission mechanism that facilitates strain diversity may eventually select for a strain that will become pathogenic. OBJECTIVE: This study was performed to examine the prevalence, genotypic distribution, and mode of transmission of TTV in detail. STUDY DESIGN: Three groups of study subjects were recruited between October 1998 and January 2000 in Prague, Czech Republic. Group 1 included 152 injection drug users with liver disease; group 2 included 102 persons with liver disease who denied ever using injection drugs; group 3 included 111 prospective blood donors. TTV DNA was detected from blood by a semi-nested PCR assay, and a selected set of PCR products was genotyped by direct sequencing. Factors associated with TTV prevalence in groups 1 and 2 subjects were compared. RESULTS: TTV was detected in 15.8, 13.7, and 13.5% of Groups 1, 2, and 3 subjects, respectively (P>0.05). The most common genotype was 2 (54%), followed by 1 (13%). The prevalence of TTV viremia was nearly three times higher in persons with a present or past history of hepatitis B compared to those without (P<0.05). TTV prevalence increased proportionately with the number of lifetime sex partners in both groups (P<0.05); it was highest (32%) among non-users of injection drugs who had five or more lifetime sex partners. CONCLUSION: TTV prevalence in the Czech population is similar among blood donors, persons with liver disease, as well as in a high-risk population of injection drug users. TTV appears to be sexually transmitted.
Adolescent↗
Recombinant Mycobacterium tuberculosis protein associated with mammalian cell entry.
The ability to gain entry and resist the antimicrobial intracellular environment of mammalian cells is an essential virulence property of Mycobacterium tuberculosis. A purified recombinant protein expressed by a 1362 bp locus (mce1) in the M. tuberculosis genome promoted uptake into HeLa cells of polystyrene latex microspheres coated with the protein. N-terminus deletion constructs of Mce1 identified a domain located between amino acid positions 106 and 163 that was needed for this cell uptake activity. Mce1 contained hydrophobic stretches at the N-terminus predictive of a signal sequence, and colloidal gold immunoelectron microscopy indicated that the corresponding native protein is expressed on the surface of the M. tuberculosis organism. The complete M. tuberculosis genome sequence revealed that it contained four homologues of mce (mce1, mce2, mce3, mce4) and that they were all located within operons composed of genes arranged similarly at different locations in the chromosome. Recombinant Mce2, which had the highest level of identity (67%) to Mce1, was unable to promote the association of microspheres with HeLa cells. Although the exact function of Mce1 is still unknown, it appears to serve as an effector molecule expressed on the surface of M. tuberculosis that is capable of eliciting plasma membrane perturbations in non-phagocytic mammalian cells.
Antibodies, Bacterial↗
Identification of new repetitive element in Leptospira interrogans serovar copenhageni and its application to PCR-based differentiation of Leptospira serogroups.
A new repetitive DNA element was identified in an isolate of Leptospira interrogans serovar copenhageni from a patient in Salvador, Brazil. A Sau3A genomic library from this strain was constructed and screened for repetitive DNA elements. An insert of 438 bp (Rep1) from one library clone hybridized to multiple chromosomal DNA fragments resolved electrophoretically after digestion with BamHI, HindIII, and MfeI. A single oligonucleotide primer, designated iRepl, was designed to generate multiple PCR amplicons of various electrophoretic mobilities in a PCR typing method. The method distinguished strains belonging to the eight pathogenic and three saprophytic species of the genus Leptospira. Clinical isolates obtained during urban epidemics between 1996 and 1998 in Salvador, Brazil, were analyzed by this PCR method. Although the iRep1 primer was unable to discriminate strains among L. interrogans serovar copenhageni isolates, it was able to differentiate strains belonging to different species and serogroups of Leptospira identified in Salvador. This PCR-based method may provide a faster and less expensive alternative to serologic tests used in reference laboratories.
Animals↗