Diagnostic parameters of modified two-tier testing in European patients with early Lyme disease.

Modified two-tier testing (MTTT) for Lyme borreliosis (i.e. confirmation with an EIA instead of an immunoblot) has been shown to have improved sensitivity compared with standard two-tier testing (STTT) in samples from American patients, without losing specificity. The current study assesses the sensitivity and specificity of various algorithms of MTTT in European patients with erythema migrans (EM) as a model disease for early Lyme borreliosis, and in appropriate controls. Four different immunoassays were used in the first tier, followed by either an immunoblot or the C6-EIA, or were used as standalone single-tier test. These tests were performed on consecutively collected sera of 228 Dutch patients with physician-diagnosed EM in the setting of general practice, 231 controls from the general population, and 50 controls with potentially cross-reactive antibodies. All the variants of MTTT that were studied had significantly higher sensitivity compared with their equivalent STTT, while retaining comparable specificity. Within the MTTT algorithms, classifying equivocal results as positive yielded better diagnostic parameters than classifying equivocal results as negative. The best diagnostic parameters were found using the Enzygnost-2 assay in the first tier, followed by a C6-ELISA in the second tier (sensitivity 77.6%, 95% CI 71.7-82.9; specificity 96.1%, 95% CI 92.7-98.2). This algorithm performed significantly better than the equivalent STTT algorithm in terms of sensitivity (p < 0.001), while maintaining comparable specificity (population controls p = 0.617). Our results show that MTTT can be a useful tool for the serodiagnosis of European patients with early Lyme borreliosis.

Evaluation of eight enzyme immunoassays for detection of immunoglobulin G against Helicobacter pylori.

Eight commercial enzyme-linked immunosorbent assays (ELISAs) were used to test sera taken from 102 patients in whom Helicobacter pylori infection status had been determined by means of biopsy culture, PCR, histology, and urease production and by 13C urea breath test. By those means, 61 patients had been found to be infected. Assays were compared by receiver operating characteristic analysis. Sensitivities ranged from 86 to 98%; specificities ranged from 83 to 98%. In a group consisting of the assays by Bio-Whittaker, Meddens Biotech, Orion (Pyloriset EIA G, new version), and Enteric Products, Inc. (HM Cap), differences in performance were not statistically significant. Sensitivities in this group ranged from 93 to 98%; specificities ranged from 95 to 98%. Assays from this group may be useful in addition to biopsy-based methods in diagnosing H. pylori infection.

Micromorphometrical analysis of rodent related (SPF) and unrelated (human) gut microbial flora in germfree mice by digital image processing.

Digital image processing (DIP) of bacterial smears is a new method of analysing the composition of the gut microbial flora. This method provides the opportunity to compare and evaluate differences in the complex highly concentrated anaerobic fraction of gut microbial flora, based on micromorphological differences. There is ample evidence that this fraction can be characterized as related or unrelated to the host organism by its immunogenicity. In this study germfree ND2 mice were associated with either related (rodent) SPF microflora (SPF-MF) or unrelated human MF (HUM-MF). DIP analysis was performed on original SPF-MF and HUM-MF and on the faeces of ex-germfree mice 4 weeks after association. The micromorphological pattern of highly concentrated anaerobic bacteria in faeces of HUM-MF associated ex-germfree mice was significantly different from SPF-MF associated counterparts with regard to the scores for elongation (P < 0.01) and morphological variety (P < 0.05). Moreover, gross morphological variability was present between individual HUM-MF associated mice but not between individual SPF-MF associated animals. No differences were found between original SPF and HUM-MF. The data are discussed with regard to differences in the presence of (non-)immunogenic bacteria and the ability for related and unrelated flora to colonize the murine gut. This study provides evidence that murine host specificity of microbial flora may not only be reflected in the number of non-immunogenic bacteria but also in the micromorphological pattern of highly concentrated anaerobic bacteria in faeces measured by DIP analysis.

DATAPLOT: a graphical display package for bacterial morphometry and fluorimetry data.

A microbiological image processing system has been developed, which is used to determine both morphometry and fluorimetry data on (mainly faecal) bacteria. A special program for display of these data has been developed for this system. Besides probability density estimates of any one or two parameters and scatter plots of any two or three parameters in the distribution, it can generate scatter plots using the actual bacterial shapes, rather than arbitrary symbols. A particularly powerful feature of the program is the possibility to overlay a contour plot of the probability density distribution of the bacteria in parameter space, with a scatter plot of the actual bacterial shapes involved. In this paper, the program's organization, options, and the algorithms used are discussed.

Morphometrical parameters of gut microflora in human volunteers.

The morphology of faecal microflora of nine healthy human volunteers was studied by digital image analysis of microscopic slides. Weekly specimens were collected during an 8-week period. Seven morphometrical parameters were derived: the means and medians of components 1, 2 and 3, and morphometrical entropy. Statistically significant differences among subjects were found for means of components 1 and 2, medians of components 1, 2 and 3, and entropy. The stability in normal circumstances provides an excellent basis for the detection of pathological change in gut flora balance.

Effects of ceftriaxone on faecal flora: analysis by micromorphometry.

In order to elucidate the effect of ceftriaxone therapy on the morphology of gut microflora, 11 human volunteers were treated with ceftriaxone, 1 g daily, given intramuscularly in one dose. Treatment continued for 5 days. Faecal microflora was analysed by digital image processing before, during and after the treatment period. We derived simple numerical parameters which describe the morphologic composition of the flora. They were significantly influenced by the antibiotic, and returned to their baseline values more than 7 days after treatment was stopped. The procedure holds promise for clinical application.

Short-chain fatty acid and organic acid concentrations in feces of healthy human volunteers and their correlations with anaerobe cultural counts during systemic ceftriaxone administration.

Short-chain fatty acid (SCFA) and organic acid concentrations and (an)aerobe cultural counts were determined simultaneously in fresh feces from 10 human volunteers before, during, and after 5 days of single-dose ceftriaxone administration. The changes and the correlation coefficients between these variables were tested to be significant (P less than 0.05). to investigate whether significant changes in the intestinal microflora could be predicted by significant changes in SCFA/organic acid concentrations. Multivariate analysis was used with the same objective. Significant decreases in most SCFA, total SCFA (0.01 less than P less than 0.02), and pyruvic acid (0.02 less than P less than 0.05) concentrations in feces were found on day 3 of treatment and also on day 5, with the exception of the pyruvic acid concentrations. Lactic acid concentrations, however, were significantly increased (0.01 less than P less than 0.02) on day 5. All anaerobic cultural counts were significantly decreased (P less than 0.05) on days 3 and 5 of treatment except those for Clostridium difficile agar, which were only decreased on day 5. Highly significant correlations were found on day 3 between the total anaerobe counts (P less than 0.001), the Bacteroides counts (P less than 0.001), and propionic acid, of which the latter had a predictive value during treatment. The results of multivariate analysis were significant but did not have a predictive value. From this study we can conclude that propionic acid concentrations during ceftriaxone treatment in healthy volunteers have a predictive value for the Bacteroides cultural counts. The results of this study must be confirmed by those from patients treated with ceftriaxone.