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  • 1
    Electronic Resource
    Electronic Resource
    Borrelia burgdorferi-associated cutaneous marginal zone lymphoma: a clinicopathological study of two cases illustrating the temporal progression of B. burgdorferi-associated B-cell proliferation in the skin (2000)
    Oxford, UK : Blackwell Science Ltd
    Histopathology 37 (2000), S. 0 
    Details
    ISSN: 1365-2559
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: A relationship between Borrelia burgdorferi and primary cutaneous B-cell lymphoma (PCBCL) has recently been confirmed following demonstration of the organism in lesional skin of patients with PCBCL. We report herein two cases of B. burgdorferi-associated PCBCL which strengthen this association by demonstrating the organism in cutaneous B-cell infiltrates present at sites in which PCBCL subsequently developed.All studies were performed on formalin-fixed paraffin-embedded tissues. These were examined by routine light microscopy and immunohistochemically by a standard streptavidin–biotin–complex technique. Genotypic studies were also undertaken using semi-nested polymerase chain reaction (PCR) for immunoglobulin heavy chain gene rearrangement, and nested PCR for B. burgdorferi flagellin gene. Both patients presented with erythematous skin lesions, biopsy of which showed dense perivascular infiltrates comprising small T-lymphocytes and collections of B-blasts. Primary cutaneous marginal zone lymphoma (MZL) developed subsequently in both cases at the same site. PCR for B. burgdorferi flagellin gene was positive in the perivascular lymphocytic infiltrates and the succeeding lymphomas in both patients.These results show that, at least in some instances, PCBCL arises from chronically stimulated lymphoid tissue acquired in the skin in response to B. burgdorferi infection. This may have significant therapeutic implications and warrant further studies on the extent of this association.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-2559.2000.01003.x
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  • 2
    Electronic Resource
    Electronic Resource
    Cell-Culture System for Continuous Production of Toxoplasma gondii Tachyzoites (1999)
    Springer
    European journal of clinical microbiology & infectious diseases 18 (1999), S. 879-884 
    Details
    ISSN: 1435-4373
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The aim of this study was to identify a sustainable cell line and culture method that could continuously provide a sufficient quantity of Toxoplasma gondii tachyzoites to serve the needs of a general hospital laboratory. Three continuous cell lines (HeLa, LLC and Vero) and three cell-culture methods (culture in conventional flasks, culture in membrane-based flasks and an automated culture system) were investigated. In multiplicity-of-infection and time-course experiments, HeLa was the cell line of choice. Harvests from HeLa cells had significantly higher tachyzoite yields than those from LLC cells (P〈0.00005) or Vero cells (P〈0.05). Membrane-based flasks gave higher yields (6.15×106 tachyzoites/ml) than conventional flasks (1–2×106 tachyzoites/ml) initially, but these were not sustained. The automated cell-culture system was unsuitable for parasite culture. Continuous passage in 25 cm2 flasks was successful, yielding 1×106 tachyzoites/ml; viability exceeded 90% after 96–120 h of infection throughout 38 passes, during which time the viability improved and the time to harvest became more consistent. Toxoplasma gondii grown in continuous culture in HeLa cells can provide a regular supply of viable tachyzoites. Demonstration that HeLa-derived tachyzoites could be used for the dye test confirms the potential of this in vitro system for use in general hospital laboratories.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s100960050423
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  • 3
    Electronic Resource
    Electronic Resource
    Evidence-Based Diagnosis of Lyme Disease (1999)
    Springer
    European journal of clinical microbiology & infectious diseases 18 (1999), S. 484-489 
    Details
    ISSN: 1435-4373
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The aim of this study was to make an evidence-based comparison of four commercial enzyme immunoassays (EIAs) (Serion Classics, Sigma Diagnostics, Cambridge Biotech and ICN Diagnostics) and an in-house enzyme immunoassay (EIA) in order to select the most appropriate screening assay for diagnosis of Lyme disease. Borrelia burgdorferi sensu stricto cultured in BSK-H medium was used to develop the in-house assay. Escherichia coli antigen (0.9 mg/ml) was included in the serum diluent to reduce non-specific background. Comparison of the number of tests needed to diagnose (i.e. to indicate a positive result) and the cost per positive diagnosis for the five assays was made using a panel of 176 Western blot-characterised sera. The Cambridge Biotech and Sigma assays had the highest sensitivity but poorer specificity, whereas the Serion and ICN assays had highest specificity but poorer sensitivity. The in-house assay had average sensitivity and specificity, the number of tests needed to diagnose being 2.32 compared to 1.92 for Serion, 2.17 for ICN, 2.5 for Sigma and 2.7 for Cambridge Biotech. In a diagnostic protocol that uses an EIA as screening test, with confirmation by Western blot, a good balance of sensitivity and specificity is essential. The in-house assay was the most cost-effective (lowest cost per positive diagnosis), and is probably the best option for specialist laboratories in Europe.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s100960050328
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  • 4
    Electronic Resource
    Electronic Resource
    Evidence-Based Diagnosis of Toxoplasma Infection (2000)
    Springer
    European journal of clinical microbiology & infectious diseases 19 (2000), S. 829-833 
    Details
    ISSN: 1435-4373
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The aim of this study was to compare the performance of one in-house and four commercially available toxoplasma assays with the Sabin-Feldman dye test. One hundred fifty-seven routine sera and 20 potentially cross-reactive sera were tested blindly using four commercial assays: Abbott AxSym IgG (Abbott Laboratories, UK), Captia Select Toxo-G (Trinity Biotech, UK), Toxreagent 'Eiken' (Eiken Chemical, Japan) and Toxolatex Fumouze (Fumouze Laboratoires, France); an in-house IgG and IgM enzyme immunoassay (EIA); and the gold standard Sabin-Feldman dye test. The sensitivity, specificity and the values using the formulae for numbers needed to diagnose (NND) and the cost per positive diagnosis (CPPD) were calculated for each assay. These formulae use the sensitivity and specificity of the assay to allow for evidence-based comparisons between assays. The NND values for the in-house IgG EIA, AxSym, Eiken and Fumouze latex kits were similar (1.21–1.24), whereas the Captia yielded the poorest value (1.33). The in-house EIA IgG had the lowest CPPD value (£0.57/$0.91), and the Fumouze and Eiken latex kits had the lowest CPPD values for commercial assays (£1.42/$2.27 and £1.81/$2.90, respectively). Both assays were simple and straightforward to use. Specialist laboratories should opt to use in-house assays, as they were most cost-effective. Although nonspecialist laboratories could use commercial assays, specimens from immunocompromised patients and patients with ocular disease should be forwarded to specialist laboratories without prior testing.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s100960000378
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