1. |
- Helldal, Lisa, et al.
(författare)
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Evaluation of MLVA for epidemiological typing and outbreak detection of ESBL-producing Escherichia coli in Sweden
- 2017
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Ingår i: Bmc Microbiology. - : Springer Science and Business Media LLC. - 1471-2180. ; 17
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Tidskriftsartikel (refereegranskat)abstract
- Background: To identify the spread of nosocomial infections and halt outbreak development caused by Escherichia coli that carry multiple antibiotic resistance factors, such as extended-spectrum beta-lactamases (ESBLs) and carbapenemases, is becoming demanding challenges due to the rapid global increase and constant and increasing influx of these bacteria from the community to the hospital setting. Our aim was to assess a reliable and rapid typing protocol for ESBL-E. coli, with the primary focus to screen for possible clonal relatedness between isolates. All clinical ESBL-E. coli isolates, collected from hospitals (n = 63) and the community (n = 41), within a single geographical region over a 6 months period, were included, as well as clinical isolates from a polyclonal outbreak (ST131, n = 9, and ST1444, n = 3). The sporadic cases represented 36 STs, of which eight STs dominated i.e. ST131 (n = 33 isolates), ST648 (n = 10), ST38 (n = 9), ST12 and 69 (each n = 4), ST 167, 405 and 372 (each n = 3). The efficacy of multiple-locus variable number tandem repeat analysis (MLVA) was evaluated using three, seven or ten loci, in comparison with that of pulsed-field gel electrophoresis (PFGE) and multi locus sequence typing (MLST). Results: MLVA detected 39, 55 and 60 distinct types, respectively, using three (GECM-3), seven (GECM-7) or ten (GECM-10) loci. For GECM-7 and -10, 26 STs included one type and eleven STs each included several types, the corresponding numbers for GECM-3 were 29 and 8. The highest numbers were seen for ST131 (7,7 and 8 types, respectively), ST38 (5,5,8) and ST648 (4,5,5). Good concordance was observed with PFGE and GECM-7 and -10, despite fewer types being identified with MLVA; 78 as compared to 55 and 60 types. The lower discriminatory power of MLVA was primarily seen within the O25b-ST131 lineage (n = 34) and its H30-Rx subclone (n = 21). Epidemiologically unrelated O25b-ST131 isolates were clustered with O25b-ST131 outbreak isolates by MLVA, whereas the ST1444 outbreak isolates were accurately distinguished from unrelated isolates. Conclusion: MLVA, even when using only three loci, represents an easy initial typing tool for epidemiological screening of ESBL-E. coli. For the ST131-O25b linage, complementary methods may be needed to obtain sufficient resolution.
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2. |
- Helldal, Lisa, et al.
(författare)
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Increasing prevalence of ESBL but not AmpC in Escherichia coli and Klebsiella pneumoniae, in Göteborg, 2004-2008
- 2009
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Ingår i: Scandinavian Society for Antimicrobial Chemotherapy - 2009, September 3, Tromsø, Norway.
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- Introduction: The increasing prevalence of transferable broad-spectrum resistance to beta-lactams, such as Extended Spectrum Beta Lactamases (ESBL) and AmpC, is troublesome, since they confer resistance to cephalosporins and penicillins, and often also are associated with additional resistance. Materials and methods: Resistance for E. coli and Klebsiella pneumoniae isolated from urine (~10.000 isolates/year) and blood (~xxx isolates/year), during 2004-2008 were determined. Cephalosporin resistant isolates were examined for presence of ESBL with a double-disk-assay using clavulanic acid as inhibitory substance. Cefoxitine-resistant strains were analyzed for presence of AmpC with a second double-disk-assay using cloxacillin as inhibitory substance. Positive strains where further tested with PCR assays for ESBL and plasmid AmpC. Results: During 2004-2008 presence of ESBL increased from 0,3–1,5% in urinary and 0,0–1,4 % in blood E. coli. For Klebsiella the corresponding figures were 0,08–0,68% and 0%, respectively. For ESBL-producing E. coli, 60–80% were resistant also to quinolones and trimetoprime. In 2008, the vast a majority of the ESBL-isolates carried CTX-M subtype 1. Approximately 50 % is community-acquired isolates. 0,15-0,23% of the urinary E. coli-strains had phenotypical characteristics indicating AmpC-production. Presence of plasmid-mediated AmpC will be tested. Approximately 50% of these were multidrug resistant. In blood E coli isolates as well as in Klebsiella from urine and blood AmpC was rarely detected (0-2 isolates/year). Discussion and Conclusion: There is a steady increase in ESBL-producing bacteria in our region. However, the frequency of isolates with AmpC is still low. In addition, a majority of these strains are multidrug resistant which is particularly alarming.
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3. |
- Helldal, Lisa, et al.
(författare)
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Shift of CTX-M genotypes has determined the increased prevalence of extended-spectrum beta-lactamase-producing Escherichia coli in south-western Sweden
- 2013
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Ingår i: Clinical Microbiology and Infection. - : Elsevier BV. - 1198-743X .- 1469-0691. ; 19:2
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Tidskriftsartikel (refereegranskat)abstract
- The prevalence of Escherichia coli producing extended-spectrum β-lactamases (ESBLs) markedly increased during 2004-2008 in south-western Sweden, with a greater increase in urinary isolates in hospitals (0.2-2.5%) than in the community (0.2-1.6%). ESBLs of genotype CTX-M predominated, with a significant (p<0.02) shift from the CTX-M-9 to CTX-M-1 phylogroup occurring among urinary ESBL-producing E.coli isolated early (n=41) as compared with late (n=221) in the study period. The increase in ESBL-producing E.coli was polyclonal, and only partly attributable to an increase (0-24%) in the number of O25b-ST131 isolates carrying CTX-M-15. The increase was prominent in men and in elderly patients, and warrants continued surveillance.
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4. |
- Helldal, Lisa, et al.
(författare)
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Shifts in Extended-Spectrum Beta-Lactamase types with increasing prevalence of Escherichia coli producing ESBL
- 2010
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Ingår i: 20th European Congress of Clinical Microbiology and Infectious Diseases (ECCMID), Vienna, Austria.
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- Objectives: Contrary to other multidrug-resistant pathogens, the prevalence of bacteria producing extended-spectrum beta-lactamase (ESBL) is increasing rapidly in Sweden. In Europe, ESBL of CTX-M-, TEM-, OXA- and SHV-types are generally associated with E. coli infections, CTX-M being the most predominant. We have investigated how the prevalence of these types has changed during the last five years in the low endemic setting of western Sweden. Methods: Yearly resistance in urinary (approximately 10,000 isolates/year) and blood (approximately 250 isolates/year) E. coli during 2004-2008 were determined. Cephalosporin-resistant isolates were screened for ESBL, using a double-disk assay with clavulanic acid as the inhibitory agent. All ESBL-E. coli isolated in the region during the periods Sept 2003-April 2005 (n=46) and April 2008-March 2009 (n=256) were typed by multiplex-PCR, detecting CTX-M, TEM, OXA and SHV. CTX-M-positive isolates were sub-typed by real time Q-PCR for CTX-M-1, CTX-M-2 and CTXM-9 groups. Results: During 2004-2008, ESBL-producing E. coli strains increased from 0.3-1.5% in urinary and 0-1.4% in blood isolates. Resistance to quinolones and trimethoprim was observed in 60-80% of strains, as compared to less than 8% in non-ESBL-producing E. coli. The majority of the ESBL-E. coli strains possessed the CTX-M gene-type, increasing from 78% (36/46) in 2003-2005 to 93% (238/256) in 2008-2009. Between these time-periods, a marked shift occurred in the distribution of CTX-M types, in that strains with the CTX-M-9 group decreased from 42% (15/36) of isolates to 21% (51/238, p=0.01) and, simultaneously, strains with the CTX-M-1 group increased from 58% (21/36) to 78% (185/238, p= 0.02). Furthermore, strains of CTX-M-type exhibiting also TEM- and/or OXA increased to comprise 86% of cases, as compared to 75% previously. Similar trends were seen for community and hospital detected isolates and with no differences associated with age in affected patients. Conclusion: A steady increase in multidrug-resistant ESBL-E. coli, possessing the genes for multiple ESBL-types, was observed in western Sweden, contrary to the patterns of other multidrug-resistant bacteria. As ESBL has increased during the five-year study period, we detected a shift in the prevalence of ESBL-types, currently dominated by the CTX-M-1 group. These observations suggest that a novel ESBL-producing E. coli clone may have emerged in the area, which will be further investigated and presented.
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5. |
- Karami, Nahid, 1959, et al.
(författare)
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Sub-Typing of Extended-Spectrum-beta-Lactamase-Producing Isolates from a Nosocomial Outbreak: Application of a 10-Loci Generic Escherichia coli Multi-Locus Variable Number Tandem Repeat Analysis
- 2013
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Ingår i: Plos One. - : Public Library of Science (PLoS). - 1932-6203. ; 8:12
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Tidskriftsartikel (refereegranskat)abstract
- Extended-spectrum beta-lactamase producing Escherichia coli (ESBL-E. coli) were isolated from infants hospitalized in a neonatal, post-surgery ward during a four-month-long nosocomial outbreak and six-month follow-up period. A multi-locus variable number tandem repeat analysis (MLVA), using 10 loci (GECM-10), for 'generic' (i.e., non-STEC) E. coli was applied for sub-species-level (i.e., sub-typing) delineation and characterization of the bacterial isolates. Ten distinct GECM-10 types were detected among 50 isolates, correlating with the types defined by pulsed-field gel electrophoresis (PFGE), which is recognized to be the 'gold-standard' method for clinical epidemiological analyses. Multi-locus sequence typing (MLST), multiplex PCR genotyping of bla(CTX-M), bla(TEM), bla(OXA) and bla(SHV) genes and antibiotic resistance profiling, as well as a PCR assay specific for detecting isolates of the pandemic O25b-ST131 strain, further characterized the outbreak isolates. Two clusters of isolates with distinct GECM-10 types (G06-04 and G07-02), corresponding to two major PFGE types and the MLST-based sequence types (STs) 131 and 1444, respectively, were confirmed to be responsible for the outbreak. The application of GECM-10 sub-typing provided reliable, rapid and cost-effective epidemiological characterizations of the ESBL-producing isolates from a nosocomial outbreak that correlated with and may be used to replace the laborious PFGE protocol for analyzing generic E. coli.
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6. |
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7. |
- Åhrén, Christina, et al.
(författare)
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Outbreak with ESBL (CTX-M)-producing Escherichia coli in a pediatric surgical ward
- 2009
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Ingår i: Scandinavian Society for Antimicrobial Chemotherapy 2009, September 3, Tromsø, Norway.
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- Introduction: Nosocomial outbreaks with ESBL-producing bacteria in Scandinavia are still rare. In a surgical ward carrying mainly for small children with congenital gastrointestinal disorders spread with ESBL-producing bacteria most likely had been ongoing for three months when we detected the outbreak in December 2008. Five children with ESBL-infections had been cared for since September. Four had septicaemia as compared to no E.coli isolated in blood the previous year. Materials and methods: ESBL-detection has been performed according to routine methods. Positive isolates from patients hospitalised in the ward since 2007 were typed with pulse field gel electrophoresis (PFGE) and the PhP-phenplate method. Results: Altogether 125/169 children hospitalised during September-December were screened and 23 were positive for ESBL-producing bacteria (~50 available isolates), of which15 were only positive in stool. They had been hospitalised for a few days to several months. Four children probably constituted the infection pool. Twenty children carried the likely outbreak strain, but ESBL-producing E coli with four additional PFGE-types as well as Klebsiella pneumonie of one type were identified. Each type demonstrated up to three different resistance-patterns against trimetoprim, ciprofloxacin and tobramycin. Six children had multiple types. Discussion and Conclusion: Spread of ESBL-producing bacteria may go undetected for a long time when only clinical isolates are available. By comparing resistance pattern we missed this outbreak by more than a month. PFGE has been an invaluable tool in the investigation. Through cohorting, enforced hygiene routines, including food handling, for personnel, parents and siblings no new child has been infected (uninfected children are screened twice weekly) since the outbreak was detected.
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8. |
- Abdeldaim, Guma, 1969-, et al.
(författare)
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Multiplex quantitative PCR for detection of lower respiratory tract infection and meningitis caused by Streptococcus pneumoniae, Haemophilus influenzae and Neisseria meningitidis
- 2010
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Ingår i: BMC Microbiology. - : Springer Science and Business Media LLC. - 1471-2180. ; 10, s. 310-
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Tidskriftsartikel (refereegranskat)abstract
- Background. Streptococcus pneumoniae and Haemophilus influenzae cause pneumonia and as Neisseria meningitidis they are important agents of meningitis. Although several PCR methods have been described for these bacteria the specificity is an underestimated problem. Here we present a quantitative multiplex real-time PCR (qmPCR) for detection of S. pneumoniae (9802 gene fragment), H. influenzae (omp P6 gene) and N. meningitidis (ctrA gene). The method was evaluated on bronchoalveolar lavage (BAL) samples from 156 adults with lower respiratory tract infection (LRTI) and 31 controls, and on 87 cerebrospinal fluid (CSF) samples from meningitis patients. Results. The analytical sensitivity was not affected by using a combined mixture of reagents and a combined DNA standard (S. pneumoniae/H. influenzae/N. meningitidis) in single tubes. By blood- and BAL-culture and S. pneumoniae urinary antigen test, S. pneumoniae and H. influenzae were aetiological agents in 21 and 31 of the LTRI patients, respectively. These pathogens were identified by qmPCR in 52 and 72 of the cases, respectively, yielding sensitivities and specificities of 95% and 75% for S. pneumoniae, and 90% and 65% for H. influenzae, respectively. When using a cut-off of 105 genome copies/mL for clinical positivity the sensitivities and specificities were 90% and 80% for S. pneumoniae, and 81% and 85% for H. influenzae, respectively. Of 44 culture negative but qmPCR positive for H. influenzae, 41 were confirmed by fucK PCR as H. influenzae. Of the 103 patients who had taken antibiotics prior to sampling, S. pneumoniae and H. influenzae were identified by culture in 6% and 20% of the cases, respectively, and by the qmPCR in 36% and 53% of the cases, respectively. In 87 CSF samples S. pneumoniae and N. meningitidis were identified by culture and/or 16 S rRNA in 14 and 10 samples and by qmPCR in 14 and 10 samples, respectively, giving a sensitivity of 100% and a specificity of 100% for both bacteria. Conclusions. The PCR provides increased sensitivity and the multiplex format facilitates diagnosis of S. pneumoniae, H. influenzae and N. meningitidis and the assay enable detection after antibiotic treatment has been installed. Quantification increases the specificity of the etiology for pneumonia.
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9. |
- Alaridah, Nader, et al.
(författare)
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Transmission dynamics study of tuberculosis isolates with whole genome sequencing in southern Sweden
- 2019
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Ingår i: Scientific Reports. - : Nature Publishing Group. - 2045-2322. ; 9
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Tidskriftsartikel (refereegranskat)abstract
- Epidemiological contact tracing complemented with genotyping of clinical Mycobacterium tuberculosis isolates is important for understanding disease transmission. In Sweden, tuberculosis (TB) is mostly reported in migrant and homeless where epidemiologic contact tracing could pose a problem. This study compared epidemiologic linking with genotyping in a low burden country. Mycobacterium tuberculosis isolates (n = 93) collected at Scania University Hospital in Southern Sweden were analysed with the standard genotyping method mycobacterial interspersed repetitive units-variable number tandem repeats (MIRU-VNTR) and the results were compared with whole genome sequencing (WGS). Using a maximum of twelve single nucleotide polymorphisms (SNPs) as the upper threshold of genomic relatedness noted among hosts, we identified 18 clusters with WGS comprising 52 patients with overall pairwise genetic maximum distances ranging from zero to nine SNPs. MIRU-VNTR and WGS clustered the same isolates, although the distribution differed depending on MIRU-VNTR limitations. Both genotyping techniques identified clusters where epidemiologic linking was insufficient, although WGS had higher correlation with epidemiologic data. To summarize, WGS provided better resolution of transmission than MIRU-VNTR in a setting with low TB incidence. WGS predicted epidemiologic links better which could consolidate and correct the epidemiologically linked cases, avoiding thus false clustering.
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10. |
- Berntsson, Matilda, 1968, et al.
(författare)
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Viral and bacterial aetiologies of male urethritis: findings of a high prevalence of Epstein-Barr virus
- 2010
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Ingår i: International Journal of STD & AIDS. - 0956-4624. ; 21:3, s. 191-194
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Tidskriftsartikel (refereegranskat)abstract
- Male urethritis is one of the most common sexually transmitted infections (STIs). However, the aetiology is still unclear in many cases. In this study the prevalences of Epstein-Barr virus (EBV), herpes simplex virus type 1 (HSV-1), HSV-2, cytomegalovirus (CMV), adenovirus, Chlamydia trachomatis, Mycoplasma genitalium and Ureaplasma urealyticum (including subtyping) were investigated. Samples from 112 male STI attendants with microscopically verified urethritis and from a control group of 103 men without clinical or microscopic signs of urethritis were analysed. Prevalences in the urethritis group compared with the controls were as follows: EBV 21%, 6% (P < 0.01); C. trachomatis 15%, 3% (P < 0.01); M. genitalium 6%, 1% (P = 0.067) and U. urealyticum 10%, 10% (ns). The results for HSV-1, HSV-2, CMV and adenovirus were negative in patients, and therefore not analysed in the controls. EBV was shown to be an independent predictor of urethritis and may play a role in its pathogenesis.
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