Biomedical subjects

Göran Kronvall

Publications and source records attributed to Göran Kronvall.

At least 19 recordsLinked to original sources

PubMed2006

Establishing clonal relationships between VIM-1-like metallo-beta-lactamase-producing Pseudomonas aeruginosa strains from four European countries by multilocus sequence typing.

Ten multidrug-resistant Pseudomonas aeruginosa strains producing VIM-1-like acquired metallo-beta-lactamases (MBLs), isolated from four European countries (Greece, Hungary, Italy, and Sweden), were analyzed for genetic relatedness by several methodologies, including fliC sequence analysis, macrorestriction profiling of genomic DNA by pulsed-field gel electrophoresis (PFGE), random amplification of polymorphic DNA (RAPD), and multilocus sequence typing (MLST). The four approaches yielded consistent results overall but showed different resolution powers in establishing relatedness between isolates (PFGE>RAPD>MLST>fliC typing) and could usefully complement each other to address issues in the molecular epidemiology of P. aeruginosa strains producing acquired MBLs. In particular, the recently developed MLST approach was useful in revealing clonal relatedness between isolates when this was not readily apparent using RAPD and PFGE, and it suggested a common ancestry for some of the VIM-1-like MBL-positive P. aeruginosa strains currently spreading in Europe. The MBL producers belonged in three clonal complexes/burst groups (BGs). Of these, one corresponded to the previously described BG4 and included serotype O12 strains from Hungary and Sweden, while the other two were novel and included serotype O11 or nonserotypable strains from Greece, Sweden, and/or Italy. Comparison of the integrons carrying blaVIM-1-like cassettes of various isolates revealed a remarkable structural heterogeneity, suggesting the possibility that multiple independent events of acquisition of different blaVIM-containing integrons had occurred in members of the same clonal lineage, although a contribution of integrase-mediated cassette shuffling or other recombination mechanisms during the evolution of similar strains could also have played a role in determining this variability.

Bacterial Proteins↗

PubMed2006

High diversity of group A streptococcal emm types among healthy schoolchildren in Ethiopia.

BACKGROUND: Although the prevalence of rheumatic heart disease in Ethiopia is one of the highest in the world, the epidemiology of group A streptococci (GAS) in this country is little known. GAS typing is a hallmark of both the epidemiology and understanding of diseases caused by these organisms. We have therefore conducted this study to investigate the emm (M-protein gene) type distribution of GAS carriers among Ethiopian schoolchildren. METHODS: In the present study, we performed emm typing of 82 GAS isolates collected from the throats of healthy schoolchildren (6-14 years of age) residing in 3 different urban sites in Ethiopia: Addis Ababa, Gondar, and Dire Dawa. RESULTS: We report high diversity of GAS isolates recovered from healthy schoolchildren. Eighty-two isolates represented 43 different sequence types. Thirteen newly described subtypes were detected in this study. Of the emm types prevalent in the study communities, 46% were not included in the 26-valent GAS vaccine. CONCLUSIONS: The high diversity of emm types encountered within 3 months of collection suggest that production of a vaccine candidate based on the M-protein amino termini appears to be impractical for this population. We suggest that investigations of other vaccine candidates, including the C5a peptidase, GAS carbohydrate, and fibronectin-binding proteins, as well as conserved M-protein region vaccines, should be intensified to address the needs of this population.

Adolescent↗

PubMed2006

Epidemiological MIC cut-off values for tigecycline calculated from Etest MIC values using normalized resistance interpretation.

OBJECTIVES: To apply the normalized resistance interpretation (NRI) method to Etest MIC results which have higher precision than conventional log2 dilution MIC tests due to the inclusion of intermediate values. If successful, NRI might provide an objective tool for the definition of epidemiological MIC cut-off values. METHODS: MICs of tigecycline and other antimicrobial agents were determined for 4771 clinical isolates comprising five Gram-positive and 13 Gram-negative species or species groups using the Etest. Histograms of MIC values were constructed for each species and NRI calculations were applied to them. An upper MIC limit of 2.5 SD above the theoretical mean of the normalized distribution was used for setting the epidemiological cut-off values. RESULTS: Calculated cut-off values for wild-type strains were between 0.11 and 0.96 mg/L for Gram-positive species, and between 0.44 and 8.3 mg/L for Gram-negative species, except for Pseudomonas aeruginosa, which had a cut-off value of 450 mg/L, consistent with earlier reports on the lack of activity of tigecycline against this species. CONCLUSIONS: NRI offers an objective method for the analysis of MICs produced using Etests and the determination of epidemiological MIC cut-off values.

Anti-Bacterial Agents↗

PubMed2006

Multidrug-resistant commensal Escherichia coli in children, Peru and Bolivia.

Using a rapid screening method, we investigated the prevalence of fecal carriage of antimicrobial drug-resistant Escherichia coli in 3,174 healthy children from 4 urban settings in Peru and Bolivia. High resistance rates were observed for ampicillin (95%), trimethoprim-sulfamethoxazole (94%), tetracycline (93%), streptomycin (82%), and chloramphenicol (70%). Lower resistance rates were observed for nalidixic acid (35%), kanamycin (28%), gentamicin (21%), and ciprofloxacin (18%); resistance to ceftriaxone and amikacin was uncommon (<0.5%). In a random sample of 1,080 resistant E. coli isolates, 90% exhibited a multidrug-resistance (MDR) phenotype. The 2 most common MDR phenotypes (ampicillin/tetracycline/trimethoprim-sulfamethoxazole and ampicillin/tetracycline/trimethoprim-sulfamethoxazole/chloramphenicol) could be transferred en bloc in conjugation experiments. The most common acquired resistance genes were blaTEM, tet(A), tet(B), drfA8, sul1, sul2, and catI. These findings underscore the magnitude of the problem of antimicrobial drug resistance in low-resource settings and the urgent need for surveillance and control of this phenomenon.

Bolivia↗

PubMed2005

Extended antimicrobial resistance screening of the dominant faecal Escherichia coli and of rare resistant clones.

Fifty faecal samples from healthy adults were grown on MacConkey agar and three pink colonies were subcultured, identified to species level and their antimicrobial susceptibility determined. Forty-seven samples yielded 141 isolates of Escherichia coli that were susceptible to most antimicrobials. Resistance was noted for ampicillin (30.5%), chloramphenicol (12.1%), tetracycline (23.4%), trimethoprim (24.8%) and co-trimoxazole (22.7%). A direct faecal plating method was used for extended resistance screening with E. coli as the indicator organism. Zone breakpoints were determined using normalised resistance interpretation and gave similar susceptibility results. Eighty-eight isolates of E. coli from within the zones of inhibition revealed four times more antimicrobial resistance. Extended antimicrobial resistance screening both provides the susceptibility profile of the dominant E. coli isolate and detects greater resistance in rare isolates.

Ampicillin↗

PubMed2005

Evaluation of a rapid screening method for detection of antimicrobial resistance in the commensal microbiota of the gut.

The assessment of antimicrobial resistance among commensal bacteria is an indicator of the spread of antimicrobial resistance. Rapid screening methods for detection of antimicrobial-resistant faecal Escherichia coli directly on MacConkey plates have been successfully adopted but suffer from lack of standardisation. The purpose of this study was to evaluate a direct plating method (DPM) for detection of antimicrobial-resistant faecal E. coli and to compare it with a conventional method. Faecal samples were collected from 71 healthy children from Peru and Bolivia. In the DPM, a faecal swab was directly plated onto a MacConkey agar plate and antimicrobial disks were applied onto the seeded plate. Raw data were obtained by direct reading of the plate and were subjected to confirmatory analysis. Good concordance between the DPM and a conventional method was observed in detecting carriage of resistant E. coli, with a higher sensitivity for the DPM. Analysis of the results allowed interpretive criteria to be defined for DPM raw data. The DPM showed good sensitivity and specificity at very low cost (ten times cheaper than the conventional method) to investigate the faecal carriage of drug-resistant E. coli. It may represent a useful tool to conduct large-scale resistance surveillance studies and to monitor resistance control programmes cost effectively, particularly in low-resource countries.

Bacteriological Techniques↗

PubMed2005

Acquired sulphonamide resistance genes in faecal Escherichia coli from healthy children in Bolivia and Peru.

Antimicrobial resistance and sulphonamide resistance determinants were studied in 20 co-trimoxazole resistant Escherichia coli in faecal samples from healthy children in Bolivia and Peru. Methods used were disc diffusion susceptibility tests, PCR, sequence analysis and plasmid conjugation assays. All isolates but one were resistant to at least two different classes of antimicrobials; 19 isolates also carried at least one sul-gene. The most frequent gene was sul2 followed by sul1 and sul3, which was detected in one isolate. This is the first observation of sul3 on the American continent. In conclusion, the high prevalence of sul-genes in this material of faecal commensal E. coli isolates points to a potential role of the faecal flora in the emergence and spread of antimicrobial resistance.

Anti-Infective Agents↗

PubMed2005

Overprescribing of antibiotics to children in rural Vietnam.

100 children (aged 1-6 y), who received an antibiotic prescription after health examination in the Bavi health care system (79 children at a district hospital, 21 at a community health centre) were analysed regarding antibiotics prescribed in relation to serum C-reactive protein (CRP). A control group consisted of 35 healthy children. Children who had been treated with antibiotics within 1 week prior to the study were excluded in the community health centre and control groups. Capillary blood samples were collected and the serum CRP concentration was analysed. A questionnaire interview with the carers was performed. Elevated CRP concentrations (>10 mg/l) were detected in 17 (17%) of the study population, and only 2 had a CRP level above 25 mg/l, one 36 mg/l and the other 140 mg/l. In the control group, none of the children had elevated CRP. The most common diagnoses were acute respiratory tract infection (ARI, 55%), asthma (7%), tonsillitis (4%), and diarrhoea (4%). The average number of drugs per patient was 3.1, and 77% received vitamins and 15% corticosteroids in combination with antibiotics. A majority of children who received an antibiotic prescription based on clinical examination did not have an elevated CRP and overprescribing of antibiotics was thus indicated.

Anti-Bacterial Agents↗

PubMed2005

Worldwide distribution of Pseudomonas aeruginosa clone C strains in the aquatic environment and cystic fibrosis patients.

Highly successful bacterial clones have the ability to effectively colonize environmental niches and patients. However, the factors which determine the complex interplay between the colonization of environmental niches and patients are mainly unknown. In this study we show that Pseudomonas aeruginosa clone C strains are distributed worldwide and highly prone to infect cystic fibrosis (CF) patients in Canada, England, France and Germany. In Hanover, Germany and Vancouver, Canada, clone C strains are highly prevalent in the CF patient community, although the mechanisms of acquisition may have been different. All clone C strains showed highly related macrorestriction fragment pattern of the whole genome as visualized by pulsed-field gel electrophoresis and harboured the 102 kbp plasmid pKLC102. Comparison of three prevalent P. aeruginosa clones with different distribution between the environment and patients revealed that neither enhanced biofilm formation nor antibiotic resistance was responsible for the spread of clone C. Clone M, which was highly prevalent in the clinical environment such as sanitary facilities, lacked motility, which could explain its relatively low prevalence in CF patients. Elucidation of the mechanisms which lead to the prevalence of clone C strain in patients and the environment requires the investigation of additional phenotypes.

Biofilms↗

PubMed2005

Carbapenem resistance mechanisms in Pseudomonas aeruginosa: alterations of porin OprD and efflux proteins do not fully explain resistance patterns observed in clinical isolates.

Imipenem resistance in Pseudomonas aeruginosa is considered to be associated with loss of the porin OprD combined with activity of chromosomal beta-lactamase (AmpC), while overexpression of multidrug efflux pumps is considered to confer meropenem resistance. Carbapenem resistance can also result from production of metallo-beta-lactamases. Transcription of oprD and efflux pump genes mexB, mexY and mexF was analysed in 23 clinical isolates of P. aeruginosa by quantitative RT-PCR. oprD was sequenced in all, and mexR, regulator of efflux pump MexAB-OprM, in selected isolates. Four isolates that were imipenem susceptible had significant reduction of oprD mRNA and presence of oprD mutations causing frameshift or translational stop. In strains only resistant to imipenem no significant difference in transcription of oprD was observed between low-level and high-level resistant isolates. The differences could not be explained by either pattern of oprD mutations. Increased transcription of mexB generally correlated well with meropenem resistance. One high-level meropenem-resistant isolate showed no significant change in mexB mRNA, but sequencing confirmed presence of a nalB mutation. Furthermore, one meropenem-susceptible isolate showed significant increase in mexB transcription, but no mexR mutations. In summary, our findings indicate that the resistance patterns observed cannot be fully explained by the currently described carbapenem resistance mechanisms.

Amino Acid Sequence↗

PubMed2005

M protein gene (emm type) analysis of group A beta-hemolytic streptococci from Ethiopia reveals unique patterns.

The genetic diversity of group A streptococcal (GAS) isolates obtained in 1990 from Ethiopian children with various streptococcal diseases was studied by using emm gene sequence analysis. A total of 217 GAS isolates were included: 155 and 62 isolates from throat and skin, respectively. A total of 78 different emm/st types were detected among the 217 isolates. Of these, 166 (76.5%) belonged to 52 validated reference emm types, 26 (11.9%) belonged to 16 already recognized sequence types (st types) and 25 (11.5%) belonged to 10 undocumented new sequence types. Resistance to tetracycline (148 of 217) was not correlated to emm type. Isolation rate of the classical rheumatogenic and nephritogenic strains was low from cases of acute rheumatic fever (ARF) and acute glomerulonephritis (AGN), respectively. Instead, the recently discovered st types were overrepresented among isolates from patients with ARF (3 of 7) and AGN (9 of 16) (P < 0.01) compared to isolates from subjects with tonsillitis and from healthy carriers (10 of 57 and 16 of 90, respectively). In contrast to rheumatogenic strains from the temperate regions, more than half of the isolates from ARF (four of seven) carried the genetic marker for skin preference, emm pattern D, although most of them (six of seven) were isolated from throat. Of 57 tonsillitis-associated isolates, 16 (28%) belonged to emm pattern D compared to <1% in temperate regions. As in other reports emm patterns A to C were strongly associated with throat, whereas emm pattern D did not correlate to skin. This first large-scale emm typing report from Africa has demonstrated a heterogeneous GAS population and contrasting nature of GAS epidemiology in the region.

Antigens, Bacterial↗

PubMed2004

Molecular cloning and characterization of two Helicobacter pylori genes coding for plasminogen-binding proteins.

Helicobacter pylori binds a number of host cell proteins, including the plasma protein plasminogen, which is the proenzyme of the serine protease plasmin. Two H. pylori plasminogen-binding proteins have been described; however, no genes were identified. Here we report the use of a phage display library to clone two genes from the H. pylori CCUG 17874 genome that mediate binding to plasminogen. DNA sequence analysis of one of these genes revealed 96.6% homology with H. pylori 26695 HP0508. A subsequent database search revealed that the amino acid sequence of a lysine-rich C-terminal segment of HP0508 is identical to the C terminus of HP0863. Recombinant proteins expressed from HP0508 and HP0863 bound plasminogen specifically and in a lysine-dependent manner. We designate these genes pgbA and pgbB, respectively. These proteins are expressed by a variety of H. pylori strains, have surface-exposed domains, and do not inhibit plasminogen activation. These results indicate that pgbA and pgbB may allow H. pylori to coat its exterior with plasminogen, which subsequently can be activated to plasmin. The surface acquisition of protease activity may enhance the virulence of H. pylori.

Amino Acid Sequence↗

PubMed2004

Integrons and multidrug resistance among Escherichia coli causing community-acquired urinary tract infection in southern India.

Antimicrobial resistance genes are often clustered in integrons, genetic elements capable of recombination. There is a paucity of data on the prevalence and role of integrons from community-acquired infections in developing countries where resistance to co-trimoxazole is high. We determined the prevalence of integrons among Escherichia coli causing community-acquired urinary tract infection (UTI). Consecutive isolates of E. coli obtained from UTI of pregnant women at the Christian Medical College Hospital, Vellore, India, during 2002 were included. All isolates were tested for susceptibility to 16 antimicrobials using the disc diffusion method and for integrons of classes 1 and 2 by PCR. Of the 58 isolates tested, 28 (48.3%) were resistant to co-trimoxazole and trimethoprim. All these isolates carried integrons. Three additional isolates were sulfonamide resistant but integron negative. Class 1 integrons were present in 21 (36.2%) isolates. Resistance to ampicillin (p=0.000), nalidixic acid (p=0.001), chloramphenicol (p=0.02), tetracycline (p=0.004) and gentamicin (p=0.02) was significantly more common in isolates with integrons. DNA sequencing of two isolates with integrons showed the presence of aadA, dfr1 and dfr7 genes. This study demonstrated that integrons are widely prevalent in India and that they might play a role in multidrug resistance in E. coli from community-acquired UTI.

Community-Acquired Infections↗

PubMed2004

Detection of CTX-M-type beta-lactamase genes in fecal Escherichia coli isolates from healthy children in Bolivia and Peru.

A survey was carried out from August to November 2002 to evaluate the antimicrobial susceptibilities of fecal Escherichia coli isolates from 3,208 healthy children from four different urban areas of Latin America, two in Bolivia (Camiri and Villa Montes) and two in Peru (Yurimaguas and Moyobamba). Ceftriaxone-resistant E. coli isolates were detected in four children, one from each of the areas sampled. The isolates exhibited a multidrug-resistant phenotype, including resistance to oxyimino-cephalosporins and aztreonam, and the MICs of ceftazidime for the isolates were lower than those of cefotaxime. By PCR and sequencing, the bla(CTX-M-2) determinant was detected in three isolates and the bla(CTX-M-15) determinant was detected in one isolate (from Peru). The CTX-M-2-producing isolates belonged to three different phylogenetic groups (groups A, B2, and D), while the CTX-M-15-producing isolate belonged to phylogenetic group D. The bla(CTX-M-2) determinants were transferable to E. coli by conjugation, while conjugative transfer of the bla(CTX-M-15) determinant was not detectable. Plasmids harboring the bla(CTX-M-2) determinant exhibited similar restriction profiles, and in all of them the gene was located on a 2.2-kb PstI fragment, suggesting a genetic environment similar to that present in In35 and InS21. The findings of the present study confirm the widespread distribution of CTX-M-type beta-lactamases and underscore the role that commensal E. coli isolates could play as a potential reservoir of these clinically relevant resistance determinants. This is the first report of CTX-M-type enzymes in Bolivia and Peru and also the first report of the detection of CTX-M-15 in Latin America.

Bolivia↗

PubMed2003

Sulphonamide resistance gene sul3 found in Escherichia coli isolates from human sources.

OBJECTIVES: The aim of this study was to investigate the molecular basis of an observed increasing resistance to trimethoprim and sulphonamides despite a simultaneous decline in co-trimoxazole consumption. The distribution of sulphonamide resistance genes sul1, sul2 and the recently discovered sul3 was studied in a collection of clinical isolates of Enterobacteriaceae. METHODS: PCR with primers specific for sul1, sul2 and sul3 was used to detect the three known sulphonamide resistance genes in the isolate collection. Sequence analysis was used for confirmation of results. Restriction endonuclease digestion and conjugational transfer assays were used for plasmid analysis. RESULTS: In 64 sulphonamide-resistant isolates, 39 sul1 genes and 48 sul2 genes were detected. Twenty-five isolates carried both sul1 and sul2 and two were negative for both genes. With PCR and sequence analysis these two were shown to harbour the new sulphonamide resistance gene sul3, which was carried by different plasmids. CONCLUSIONS: Sulphonamide resistance gene sul3, which is widespread among pigs in Switzerland, has now also been identified in two different clinical isolates of Escherichia coli, located in urinary tract infections in patients in Sweden.

Anti-Infective Agents↗

PubMed2003

Analysis of parameters and validation of method for normalized interpretation of antimicrobial resistance.

The method of normalized resistance interpretation (NRI), uses the high-zone side of the susceptible peak in a zone diameter histogram as an internal calibrator to construct the real standard distribution of susceptible isolates even in the presence of resistant isolates. NRI parameters were optimized using control strain histograms from microbiology laboratories in Stockholm, Argentina, and the Philippines. A moving average based on four-zone values was slightly better than based on two-zone average values. The optimal peak adjustment from the switch position of the moving average was 1.0 for two-zone averages and 2.5 for four-zone averages. A comparison between true means and NRI-calculated means showed a highly significant correlation (R(2)=0.963). Coefficients of variation (CV), comparing the CV of the true distribution of control strain test results with the NRI calculated distribution, identified two types of aberrant histograms. NRI calculations on clinical isolates of Escherichia coli and Staphylococcus aureus from selected laboratories showed a good agreement between the local resistance interpretations with the NRI calculated levels. One type of deviation was most marked with cephalothin histograms for E. coli isolates where the regular zone breakpoints used cut through the population of susceptible strains. With proper markers for required quality of disc test results, the NRI method might be a valuable tool for both resistance surveillance and for quality control of the disc diffusion method.

Anti-Bacterial Agents↗

PubMed2003

Determination of the real standard distribution of susceptible strains in zone histograms.

The procedure for NRI, a method for normalised resistance interpretation, is presented. By means of its promising autocalibration system the method generates zone breakpoints for resistance in zone diameter histograms for bacterial species-antibiotic combinations. These breakpoints give a normalised interpretation of resistance which is independent of disc test standard chosen, disc potency, medium used, inoculum size, etc. of the individual laboratory as long as the performance shows good reproducibility and precision. The method should be further tested as a tool for setting "bacteriological breakpoints" or "epidemiological breakpoints", for calculating resistance rates for comparative purposes, e.g. in antibiotic resistance surveillance, and for quality control of the disc diffusion test.

Drug Resistance, Bacterial↗

PubMed2003

Bacteraemia in hospitalised patients with malignant blood disorders: a retrospective study of causative agents and their resistance profiles during a 14-year period without antibacterial prophylaxis.

Bacteraemia is a major cause of morbidity and mortality in patients with haematological disorders during chemotherapy-induced neutropenia. The generally reported trend during the last two decades has been a gradual replacement of Gram-negative bacilli by Gram-positive cocci as the major causes of bacteraemia in neutropenic hosts. However, data that are unaffected by the use of antibacterial prophylaxis are scarce. Our objective therefore was to study the incidence of bacteraemia with different microorganisms in a haematology centre where antibacterial prophylaxis has not been used during the years 1988-2001. A total of 1402 episodes of clinically significant bacteraemia in 927 patients were identified. All patients were treated in the haematology wards and had an underlying haematological disorder, with lymphoma, leukaemia, and myeloma dominating. There were 536 (58%) male, and 391 (42%) female patients, with a median age of 58 years. The dominating pathogens were coagulase-negative staphylococci (CNS) 17%, Escherichia coli 16%, alpha-haemolytic streptococci 12%, Staphylococcus aureus 9%, Klebsiella spp 9%, Enterococcus spp 7%, and Pseudomonas spp 5%. The only significant incidence change was an increase of E. faecium bacteraemia. The balance between Gram-negative and Gram-positive microorganisms was essentially stable over the 14-year period. The rates of antibiotic resistance were generally low and stable. Gram-negative bacteria exhibited resistance to fluoroquinolones after 1998. The 7- and 30-day mortality rates were 6.3 and 15.6%, respectively, being significantly higher in patients with bacteraemia caused by Pseudomonas aeruginosa, Stenotrophomonas maltophilia, or E. faecium.

Adolescent↗