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  • 1. Edberg, Andreas
    et al.
    Jurstrand, Margaretha
    Johansson, Eva
    Wikander, Elisabeth
    Höög, Anna
    Ahlqvist, Thomas
    Falk, Lars
    Jensen, Jørgen Skov
    Fredlund, Hans
    Örebro University, School of Health and Medical Sciences.
    A comparative study of three different PCR assays for detection of Mycoplasma genitalium in urogenital specimens from men and women2008In: Journal of Medical Microbiology, ISSN 0022-2615, E-ISSN 1473-5644, Vol. 57, no Pt 3, p. 304-309Article in journal (Refereed)
    Abstract [en]

    The aim of this study was to compare conventional 16S rRNA gene PCR, real-time 16S rRNA gene PCR and real-time Mycoplasma genitalium adhesin protein (MgPa) gene PCR as detection methods for M. genitalium infection. The study also determined the prevalence of M. genitalium in male and female patients attending a sexually transmitted infections clinic in a rural area in the west of Sweden. First void urine (FVU) and/or urethral swabs were collected from 381 men, and FVU and/or cervical swabs and/or urethral swabs were collected from 298 women. A total of 213 specimens were used in the PCR comparative study: 98 consecutively sampled specimens from patients enrolled in the prevalence study, 36 consecutively sampled specimens from patients with symptoms of urethritis and 79 specimens from patients positive for M. genitalium by real-time MgPa gene PCR in the prevalence study. A true-positive M. genitalium DNA specimen was defined as either a specimen positive in any two PCR assays or a specimen whose PCR product was verified by DNA sequencing. The prevalence of M. genitalium infection in men and women was 27/381 (7.1 %) and 23/298 (7.7 %), respectively. In the PCR comparative study, M. genitalium DNA was detected in 61/76 (80.3 %) of true-positive specimens by conventional 16S rRNA gene PCR, in 52/76 (68.4 %) by real-time 16S rRNA gene PCR and in 74/76 (97.4 %) by real-time MgPa gene PCR. Real-time MgPa gene PCR thus had higher sensitivity compared with conventional 16S rRNA gene PCR and had considerably increased sensitivity compared with real-time 16S rRNA gene PCR for detection of M. genitalium DNA. Real-time MgPa gene PCR is well suited for the clinical diagnosis of M. genitalium.

  • 2.
    Ericsson, Henrik
    et al.
    Department of Food Hygiene, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Uppsala, Sweden.
    Unnerstad, Helle
    Department of Food Hygiene, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Uppsala, Sweden.
    Mattsson, Jens G.
    Department of Parasitology, National Veterinary Institute, Uppsala, Sweden.
    Danielsson-Tham, Marie-Louise
    Department of Food Hygiene, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Uppsala, Sweden.
    Tham, Wilhelm
    Department of Food Hygiene, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Uppsala, Sweden.
    Molecular grouping of Listeria monocytogenes based on the sequence of the inlB gene2000In: Journal of Medical Microbiology, ISSN 0022-2615, E-ISSN 1473-5644, ISSN 0022-2615, Vol. 49, no 1, p. 73-80Article in journal (Refereed)
    Abstract [en]

    The major part of the gene inlB was sequenced in 24 strains of Listeria monocytogenes belonging to serovars 1/2a, 1/2b, 1/2c, 3b and 4b. A phylogenetic analysis based on the inlB nucleotide sequences showed that strains of serovars 1/2a and 1/2c were closely related, as well as those of serovars 1/2b and 3b. Strains sharing serovar 4b could be divided into two distinct groups. There were differences in amino-acid sequence between all serovars except between serovars 1/2b and 3b. Differences in amino-acid sequence were also seen within each of the serovars 1/2a and 4b. The data presented indicate that the inlB gene may be useful for typing purposes as an alternative or complement to serotyping.

  • 3.
    Jurstrand, Margaretha
    et al.
    Örebro University, Department of Clinical Medicine.
    Jensen, Jörgen Skov
    Fredlund, Hans
    Falk, Lars
    Mölling, Paula
    Detection of Mycoplasma genitalium in urogenital specimens by real-time PCR and by conventional PCR assay2005In: Journal of Medical Microbiology, ISSN 0022-2615, E-ISSN 1473-5644, Vol. 54, no 1, p. 23-29Article in journal (Refereed)
    Abstract [en]

    A real-time LightCycler PCR (LC-PCR) with hybridization probesfor detection of Mycoplasma genitalium in endocervical and firstvoid urine specimens was developed and compared to a conventionalPCR. The primers for both assays were identical and designedto amplify a 427 bp fragment of the 16S rRNA gene of M. genitalium.The LC-PCR assay had a detection limit of < 5 bacterial genomesper reaction when dilutions of genomic DNA from a type strainof M. genitalium were tested. First void urine from 398 menand first void urine and endocervical specimens from 301 womenattending an STD clinic were analysed by LC-PCR and by the conventionalPCR. Using the conventional PCR as reference, the LC-PCR hada specificity of 99.7 % and a sensitivity of 72.2 % for thedetection of M. genitalium in first void urine samples frommen. There was no significant difference in the performanceof the LC-PCR assay compared to the conventional PCR when endocervicalswabs were considered (58 and 65 %, respectively) or with aset of endocervical swab/urine specimens for which the LC-PCRassay detected 73 % of the infections (specificity = 98.6 %and sensitivity = 68.2 %) while the conventional PCR detected85 % of the infections. With female urine specimens there wasa significant difference between the two assays (38 and 73 %,respectively; P = 0.01 McNemar's test). This illustrates theneed to analyse both endocervical and urine specimens, becauseM. genitalium DNA was detected in only one of the two specimensin a great number of the M. genitalium-infected women. The lowersensitivity of the LC-PCR assay was probably caused by a combinationof inhibition and limitations regarding the amount of templateDNA. The LC-PCR assay was easy to perform and the simultaneousamplification and detection eliminated the need for furtherhandling of PCR products. With improvement in sample preparationmethods and increased volumes of the template DNA, the LC-PCRassay could be a useful routine diagnostic method.

  • 4.
    Katouli, Mohammad
    et al.
    Laboratory for Bacteriology, Microbiology and Tumorbiology Centre, Karolinska Institute, Stockholm,Sweden.
    Nettelbladt, Carl Gustaf
    Department of Surgery, Karolinska Hospital, Stockholm, Sweden.
    Muratov, Vladislaw
    Laboratory for Bacteriology, Microbiology and Tumorbiology Centre, Karolinska Institute, Stockholm,Sweden.
    Ljungqvist, Olle
    Örebro University, School of Medical Sciences. Department of Surgery, Karolinska Hospital, Stockholm, Sweden.
    Bark, Tor
    Department of Surgery, Karolinska Hospital, Stockholm, Sweden.
    Svenberg, Torgny E.
    Department of Surgery, Karolinska Hospital, Stockholm, Sweden.
    Möllby, R.
    Laboratory for Bacteriology, Microbiology and Tumorbiology Centre, Karolinska Institute, Stockholm,Sweden.
    Selective translocation of coliform bacteria adhering to caecal epithelium of rats during catabolic stress1997In: Journal of Medical Microbiology, ISSN 0022-2615, E-ISSN 1473-5644, Vol. 46, no 7, p. 571-578Article in journal (Refereed)
    Abstract [en]

    Adult conventional rats were starved for 48 h with or without haemorrhage at 24 h, and translocation of caecal coliforms to mesenteric lymph nodes (MLNs) was measured. Translocation was detected in three of 11 rats without haemorrhage, in 6 of 11 starved and sham-operated rats and in 12 of 22 rats after haemorrhage. In contrast, only one of 13 non-instrumented and fed control rats showed translocation. Translocation was associated with more coliforms adhering to caecal epithelium in rats. Coliform isolates from caecum, caecal epithelium and MLNs were characterised and grouped into different biochemical phenotypes (BPTs) by a biochemical fingerprinting method. Of 291 BPTs detected in the caecum of all rats, 108 were also found on caecal epithelium; 36 BPTs were detected in MLNs, of which 17 were not detected either in the caecum or on the caecal epithelium of the corresponding rats. One isolate from each of these 36 BPTs was selected and compared to the others. Four common (C) BPTs (i.e., C1-C4) were identified among them. Strains of C1 formed the majority of isolates from the caecum (79%), caecal epithelium(71%) and MLNs (91%). In contrast, C2-C4 had a significantly lower incidence both in the caecum and on the caecal epithelium, but not in the MLNs. These findings indicate that not all caecal coliforms adhere to the epithelium during catabolic stress and that for translocation to occur, other bacterial properties besides adhesion are needed. It is also concluded that coliforms with a low incidence in the caecum can translocate with the same efficiency as those with a high incidence.

  • 5.
    Månsson, Emeli
    et al.
    Örebro University, School of Medical Sciences. Centre for Clinical Research, Hospital of Västmanland Västerås, Västerås, Sweden.
    Hellmark, Bengt
    Örebro University, School of Medical Sciences. Örebro University Hospital. Department of Laboratory Medicine, Örebro University Hospital, Örebro, Sweden.
    Stegger, Marc
    Statens Serum Institut, Copenhagen, Denmark.
    Andersen, Paal Skytt
    Statens Serum Institut, Copenhagen, Denmark.
    Sundqvist, Martin
    Örebro University, School of Medical Sciences. Örebro University Hospital. Department of Laboratory Medicine, Örebro University Hospital, Örebro, Sweden.
    Söderquist, Bo
    Örebro University, School of Medical Sciences. Örebro University Hospital.
    Genomic relatedness of Staphylococcus pettenkoferi isolates of different origins2017In: Journal of Medical Microbiology, ISSN 0022-2615, E-ISSN 1473-5644, Vol. 66, no 5, p. 601-608Article in journal (Refereed)
    Abstract [en]

    Purpose: The aim of the study was to characterize clinical and environmental Staphylococcus pettenkoferi isolates with regard to genomic diversity and antibiotic susceptibility pattern. Repetitive-sequence-based PCR and core genome phylogenetic analysis of whole-genome sequencing (WGS) data verified the presence of distinct clades comprising closely related S. pettenkoferi isolates from different geographical locations and origins.

    Methodology: Phylogenetic relationships between 25 S. pettenkoferi isolates collected from blood cultures and intra-operative air sampling were determined by repetitive-sequence-based PCR typing and analysis of similar to 157 000 SNPs identified in the core genome after WGS. Antibiotic susceptibility testing and tests for biofilm production (microtitre plate assay) were performed.

    Results: Repetitive-sequence-based PCR as well as WGS data demonstrated the close relatedness of clinically significant blood culture isolates to probable contaminants, as well as to environmental isolates. Antibiotic-susceptibility testing demonstrated a low level of antimicrobial resistance. The mecA gene was present in two cefoxitin-resistant isolates. No isolates were found to produce biofilm.

    Conclusion: Close genomic relatedness of S. pettenkoferi isolates from different geographical locations and origins were found within clades, but with substantial genomic difference between the two major clades. The ecological niche of S. pettenkoferi remains unconfirmed, but the presence of S. pettenkoferi in the air of the operating field favours the suggestion of a role in skin flora. Identification of S. pettenkoferi in clinical samples should, in a majority of cases, most likely be regarded as a probable contamination, and its role as a possible pathogen in immunocompromised hosts remains to be clarified.

  • 6.
    Söderquist, Bo
    et al.
    Örebro University, School of Health and Medical Sciences.
    Andersson, Mira
    Nilsson, Martin
    Nilsdotter-Augustinsson, Åsa
    Persson, Lennart
    Friberg, Örjan
    Jacobsson, Susanne
    Örebro University, School of Health and Medical Sciences.
    Staphylococcus epidermidis surface protein I (SesI): a marker of the invasive capacity of S. epidermidis?2009In: Journal of Medical Microbiology, ISSN 0022-2615, E-ISSN 1473-5644, Vol. 58, no Pt 10, p. 1395-1397Article in journal (Refereed)
1 - 6 of 6
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