Campylobacter spp. are fastidious microorganisms, and their detection by culture depends on the freshness of the stool sample and the skills of the laboratory staff. To improve laboratory diagnosis, assays for the detection of specific antigens have been developed. Here, we evaluated two assays for the detection of Campylobacter spp.-specific antigens, i.e., one immunochromatographic test and one enzyme-linked immunosorbent assay (EIA), in 38 frozen Campylobacter spp.-positive specimens and prospectively in 533 fresh stool samples with a conventional enzyme immunoassay (EIA) and culture.
Both assays were positive for 36 samples with Campylobacter jejuni and one with Campylobacter coli among 38 Campylobacter spp.-positive frozen samples. One Campylobacter lari-positive sample was identified by the immunochromatographic assay (ICA) only. In a prospective study performed within the course of routine microbiology, both assays were positive for 24/25 C. jejuni culture-positive samples (positive percent agreement, 96.0% [95% CI: 78.9–100%]). ICA and EIA also were positive for 14 and 10 culture-negative samples, respectively (negative percent agreement: ICA, 97.2% [95% CI: 95.4–98.4%]; EIA, 98.0% [95% CI: 96.4–99.0%]). In conclusion, the high agreement between both antigen-detection assays and culture indicates that both assays may be initially performed followed by culture only upon a positive test result.
Authors:Ralf Ignatius, Christiane Berg, Chris Weiland, Angela Darmer, Thilo Wenzel, Marion Lorenz, Jörg Fuhrmann and Michael Müller
Stool antigen tests are recommended for the diagnosis of Helicobacter pylori infection. Here, we compared two novel assays, i.e., one enzyme immunoassay (EIA) and one immunochromatography assay (ICA), with a chemiluminescence immunoassay (CLIA) that had previously been compared with rapid urease test, histology, and urea breath test. Two hundred sixty-six frozen stool samples with defined CLIA results (42 positives, 219 negatives, and 5 samples with borderline results) collected between January and May 2018 were thawed and immediately tested by EIA, ICA, and CLIA.
In 248 samples with repeatedly positive/negative CLIA results, EIA and ICA were positive for 40 and 37 of 41 CLIA-positive samples and yielded negative results for 206 and 201 of 207 CLIA-negative samples, respectively. There was a high positive percent agreement (EIA, 97.6%; 95% confidence interval (95% CI), 86.3–100%; ICA, 90.2%; 95% CI, 76.9–96.7%), as well as a negative percent agreement between the assays (EIA, 99.5%; 95% CI, 97.0–100%; ICA, 97.1%; 95% CI, 93.7–98.8%). This was further supported by kappa values indicating very good agreement (CLIA vs. EIA, 0.971; CLIA vs. ICA, 0.857). In conclusion, both EIA and ICA comprise valuable assays for the detection of H. pylori antigen in stool samples.
Authors:Ralf Ignatius, Robert Neuber, Heike Kietzmann, Christiane Berg, Thilo Wenzel, Jörg Fuhrmann and Michael Müller
This study aimed at evaluating in outpatients an algorithm for the laboratory diagnosis of Clostridioides (Clostridium) difficile infection (CDI), i.e., enzyme immunoassays (EIAs) detecting bacterial glutamate dehydrogenase (GDH) and toxin A/B, followed by polymerase chain reaction (PCR) analyses of samples with discordant EIA results.
In total, 9802 examinations of stool samples by GDH and toxin EIAs performed in 7263 outpatients and 488 inpatients were analyzed retrospectively. Samples with discordant EIA results had been tested by a commercially available PCR assay detecting genes of the C. difficile-specific triose phosphate isomerase (tpi) and toxin B (tcdB).
Concordant EIA results (686 C. difficile-positive, 8121 negative) were observed for 8807 (89.8%; 95% CI, 89.2–90.4%) samples. Of 958 samples with discordant EIA results, 895 were analyzed using PCR and 580 of 854 GDH-positive/borderline, toxin-negative samples (67.9%; 95% CI, 64.7–71.0%) were positive for tpi and tcdB, while 274 samples (32.1%; 95% CI, 29.0–35.3%) were tcdB-negative. In contrast, 35 of 41 GDH-negative, toxin-positive/borderline samples (85.4%; 95% CI, 71.2–93.5%) were tcdB-negative. Still, 6 samples (14.6%; 95% CI, 6.5–28.8%) yielded positive PCR results for both genes.
In conclusion, around 90% of the samples were analyzed appropriately by only applying EIAs. Approximately one third of the PCR-analyzed samples were tcdB-negative; thus, patients most likely did not require CDI treatment.
Authors:Michael Fehlings, Lea Drobbe, Macarena Beigier-Bompadre, Pablo Renner Viveros, Verena Moos, Thomas Schneider, Thomas F. Meyer, Toni Aebischer and Ralf Ignatius
Direct effects of Helicobacter pylori (H. pylori) on human CD4+ T-cells hamper disentangling a possible bacterial-mediated interference with major histocompatibility complex class II (MHC-II)-dependent antigen presentation to these cells. To overcome this limitation, we employed a previously described assay, which enables assessing human antigen-processing cell function by using murine T-cell hybridoma cells restricted by human leukocyte antigen (HLA) alleles. HLA-DR1+ monocyte-derived dendritic cells were exposed to H. pylori and pulsed with the antigen 85B from Mycobacterium tuberculosis (M. tuberculosis). Interleukin-2 (IL-2) secretion by AG85Baa97-112-specific hybridoma cells was then evaluated as an integral reporter of cognate antigen presentation. This methodology enabled revealing of interference of H. pylori with the antigen-presenting capacity of human dendritic cells.