Evaluation of a New Multiparametric Microspot Array for Serodiagnosis of Lyme Borreliosis.

BACKGROUND: Laboratory diagnosis of Lyme borreliosis follows a two-step algorithm, starting with a screening assay with maximum sensitivity followed by a confirmatory assay of positive results with respect to specificity. Line immunoassays with single recombinant antigen lines are the established confirmatory test method, although result interpretation is not standardized. The present study evaluates suitability of the multiplex test SeraSpot Anti-Borrelia IgG/IgM (SeraSpot) as confirmatory test in serodiagnosis of Lyme disease. METHODS: For this retrospective study, serum samples from patients with suspicion of Lyme borreliosis were analyzed. For determination of specificity, blood donor samples as well as rheumatoid factor (RF), ANA, and EBV positive sera were investigated. The SeraSpot IgG/IgM results were compared with the recomBead Borrelia IgG/IgM (recomBead) test as alternative multiplex-based method. RESULTS: In comparison to the recomBead test the sensitivity of SeraSpot was determined 95% for IgG/IgM detection. The analysis of blood donors revealed a specificity of ≥ 96.0% for SeraSpot IgG/IgM and of 97.3% for recomBead IgG/IgM. RF positive samples tested negative by both assays (specificity of 100%). ANA and EBV antibody positive samples resulted in specificities of 90% (SeraSpo IgG/IgM) and ≥ 94% (recomBead IgG/IgM) and ≥ 91% (SeraSpot IgG/IgM) and ≥ 91% (recomBead IgG/IgM). The intra- and interassay coefficient of variation (CV) and the lot-to-lot reproducibility of the SeraSpot assay ranged between 1 - 9% for the different Borrelia IgG and IgM antigens. Potentially interfering substances (bilirubin F, bilirubin C, hemoglobin, lipid factor, and rheumatoid factor) did not influence the SeraSpot assays. CONCLUSIONS: Our evaluation data confirm that new multiplex assay generations such as SeraSpot Anti-Borrelia IgG/IgM are reliable and robust test systems suitable for application as confirmatory tests for serodiagnosis of Lyme borreliosis.

Epidemiology and morbidity of Epstein-Barr virus infection in pediatric renal transplant recipients: a multicenter, prospective study.

BACKGROUND: The epidemiology and morbidity of Epstein-Barr virus (EBV) infection in pediatric renal transplant recipients have been characterized insufficiently. METHODS: In a prospective, multicenter study among 106 pediatric kidney allograft recipients aged 11.4 ± 5.9 years, we investigated the epidemiology of EBV infection and the relationship between EBV load, EBV serology, and EBV-related morbidity (posttransplant lymphoproliferative disease [PTLD] or symptomatic EBV infection, defined as flu-like symptoms or infectious mononucleosis). RESULTS: EBV primary infection occurred in 27 of 43 (63%) seronegative patients and reactivation/reinfection in 28 of 63 (44%) seropositive patients. There was no association between the degree or duration of EBV load and EBV-related morbidity: The vast majority (17 of 18 [94%]) of patients with a high, persistent EBV load remained PTLD-free throughout a follow-up of 5.0 ± 1.3 years, while 2 of 3 (66%) patients with EBV-related PTLD exhibited only a low EBV load beforehand. Eight of 18 (44%) patients with a high, persistent EBV load remained asymptomatic during a follow-up of 5.3 ± 2.9 years. Multivariate analysis identified the EBV high-risk (D(+)/R(-)) serostatus (odds ratio [OR], 7.07; P < .05), the presence of human leukocyte antigen (HLA)-DR7 (OR, 5.65; P < .05), and the intensity of the immunosuppressive therapy (OR, 1.53; P < .01) as independent risk factors for the development of a symptomatic EBV infection. CONCLUSIONS: Presence of EBV high-risk seroconstellation, HLA-DR7, and intensity of immunosuppressive therapy are significant risk factors for a symptomatic EBV infection, whereas there is no close association between the degree or duration of EBV load and EBV-related morbidity. Clinical Trials Registration. NCT00963248.

(Val-)Ganciclovir prophylaxis reduces Epstein-Barr virus primary infection in pediatric renal transplantation.

Epstein-Barr virus (EBV) primary infection constitutes a serious risk for pediatric transplant recipients, particularly as regards the development of EBV-related post-transplant lymphoproliferative disease (PTLD). Currently, there is no established prophylactic regimen. We investigated the association between chemoprophylaxis with valganciclovir (VGCV) or ganciclovir (GCV) and the incidence of EBV viremia in EBV-naïve pediatric renal transplant recipients (R-) who had received a graft from an EBV-positive donor (D+) and are therefore at high risk of EBV primary infection. In a prospective, multicenter trial (n = 114), we compared a cohort on chemoprophylaxis (n = 20) with a similar control cohort without chemoprophylaxis (n = 8). Over the 1-year study period, antiviral prophylaxis with VGCV/GCV was associated with a significantly decreased incidence of EBV primary infection: 9/20 patients (45%) in the prophylaxis group experienced an EBV primary infection compared to 8/8 controls (100%) (P < 0.0001). Chemoprophylaxis was associated with a significantly lower EBV viral load (P < 0.001). Type or intensity of immunosuppressive therapy did not influence the occurrence of EBV primary infection or the level/persistence of EBV viral load. Chemoprophylaxis with VGCV/GCV is associated with a reduced incidence of EBV viremia in high-risk pediatric kidney allograft recipients in the first year post-transplant. (ClinicalTrials.gov number: NCT00963248).

Where have the enteric viruses gone? - Differential effects on frequent causes of infectious diarrhoea by SARS-CoV-2 pandemic lockdown measures.

BACKGROUND: Measures of distancing, wearing face/medical masks and lockdown introduced in many countries to meet the challenges of the SARS-CoV-2 pandemic have led to gross changes in the epidemiology of important infections. The observation of decline of positive norovirus tests after introduction of lockdown in Germany led us to investigate changes in the detection of major causes of diarrhoea by comparing pre-pandemic quarters (PPQ: 1Q/17 through 1Q/20) since 2017 and pandemic quarters (PQ: 2Q/20 through 1Q/21). METHODS AND SETTING: Bioscientia Laboratory Ingelheim is a large regional clinical pathology laboratory serving > 50 hospitals and > 5000 general practitioners and specialist outpatient practices located in the federal states Hesse, Rhineland-Palatinate and North Rhine-Westphalia, Germany. Antigen detection assays were used for detection of astrovirus, adenovirus, rotavirus, and Campylobacter antigen and Clostridium difficile Toxin A/B, while norovirus was detected by qualitative RT-PCR. FINDINGS: The mean positivity-ratios of norovirus, adenovirus and astrovirus assays were 3-20 fold lower in periods PQ (2Q/20 through 1Q/21) compared to PPQ (1Q/17 through 1Q/20) (p<.01). The mean positivity-ratio was lower in PQ compared to PPQ for rotavirus (p=.31), but failed to reach statistical significance, while for campylobacter antigen (p=.91) and C. difficile Toxin A/B (p=.17) the mean positivity-ratio was even higher in PQ compared to PPQ. CONCLUSIONS: Apparently, hygienic measures used to contain the SARS-CoV-2 pandemic have differential effects on incidence of diarrhoea viruses as compared to bacterial gastrointestinal agents, particularly C. difficile, which may lead to re-evaluate measures implemented against this important cause of nosocomial diarrhoea.

Adenovirus infection and treatment with cidofovir in children after liver transplantation.

In a retrospective study, serum samples from 21 pediatric liver transplant recipients were analysed by quantitative real-time PCR for ADV infection up to 24 wk after Tx. ADV DNA was detected in serum of eight children after Tx, one of whom developed life-threatening fulminant hepatitis and sepsis. None of these children were symptomatic at the time of first detection of ADV DNA in serum after Tx. Seven children with positive ADV PCR had low adenoviral loads, showed no increase in viral load and remained clinically asymptomatic in the follow-up period of 24 wk. After 10 wk under immunosuppression one child presented clinically with adenoviral sepsis and severe necrotizing hepatitis. This patient revealed a dramatic increase of ADV from baseline titers up to 1.3 x 10(9 )copies/mL serum within 10 wk after Tx. ADV was also detected in a liver biopsy of this child at 1.2 x 10(4) copies/cell and typed by sequence analysis as human ADV species C, type 6, a rarely detected ADV type and first described in a liver transplant patient. Immunosuppression was reduced in this patient immediately and the antiviral drug cidofovir administered intravenously followed by viral suppression and clinical improvement of the child.

Evaluation of a novel immunochromatographic lateral flow assay for rapid detection of OXA-48, NDM, KPC and VIM carbapenemases in multidrug-resistant Enterobacteriaceae.

PURPOSE: The global spread and increasing clinical impact of carbapenemase-producing bacteria are alarming. Rapid diagnostic techniques for carbapenemase detection are of the utmost importance to prevent delays in efficient antibiotic therapy and the control of spread in hospitals. Recently, multiplex immunochromatographic lateral flow tests (ICTs) for the fast detection of carbapenemase-producers have become commercially available. We evaluated a novel multiplex ICT for the rapid detection of OXA-48, KPC, NDM and VIM carbapenemases. METHODOLOGY: One hundred well-characterized multidrug-resistant Enterobacteriaceae were analysed by RESIST-4 (Coris, BioConcept, Gembloux, Belgium). The reference standard included confirmation at the molecular level at the German National Reference Laboratory for multidrug-resistant Gram-negative bacteria (Ruhr-University Bochum, Germany).Results/Key findings. The tested ICT was highly reliable, showing 100 % sensitivity and 100 % specificity. CONCLUSION: As it is fast, easy to perform and has few technical requirements, the ICT represents a good opportunity to improve turnaround times and patient care for the clinical microbiology laboratory.

Comparison of two commercial carbapenemase gene confirmatory assays in multiresistant Enterobacteriaceae and Acinetobacter baumannii-complex.

Multidrug-resistant Gram-negative bacilli (MDR-GNB) producing carbapenemases are increasing at an alarming speed. Rapid confirmation of carbapenemase type will be an important diagnostic step in clinical microbiology laboratories not only to reduce the risk of transmissions but also for optimising antibiotic therapy in the future. We compared diagnostic reliability of two commercially available molecular assays (Check-Direct CPE vs. AID line probe assay) for detection and typing of carbapenemase genes in 80 well-characterized isolates of MDR-GNB. Respective strains were isolated in various clinical specimens at our clinical microbiology laboratory. The reference standard included confirmation of carbapenemase-production at the molecular level at the German National Reference Laboratory for Multidrug-resistant Gram-negative bacteria (Ruhr-University Bochum, Germany). 53 Enterobacteriaceae and 27 members of the A. baumannii-complex were used in this study. The tested assays appeared highly reliable to confirm carbapenemase-producing Enterobacteriaceae (CPE) with respective sensitivities of 97.7%, but are currently unsuitable for analysis of members of the A. baumannii-complex. Both assays are easy to perform and rapid tools for confirmation and typing of the most common carbapenemase genes in Enterobacteriaceae. Implementation should be possible for any clinical microbiology laboratory with Check-Direct CPE being easier to handle and having less technological requirements.

Complicated sea urchin-induced wound infection caused by Vibrioalginolyticus and Staphylococcus lugdunensis in a 14-year-old boy.

INTRODUCTION: Wound infections with Vibrio alginolyticus, a Gram-negative bacterium found in all temperate oceans, are rarely reported. However, a rising incidence of wound infections caused by V. alginolyticus requires better knowledge about this infectious agent. CASE PRESENTATION: We report the case of a 14-year-old boy suffering from a wound infection caused by V. alginolyticus and Staphylococcus lugdunensis after stepping on a sea urchin. Despite wound debridement and antibiotic therapy with cefaclor, the lesion did not heal over several weeks. After identification of the pathogens and antibiotic-susceptibility testing, antibiotic therapy was switched to ciprofloxacin, followed by trimethoprim/sulfamethoxazole. Two months after the accident the wound was re-epithelialized. Follow up after 6 months revealed a painful scar. CONCLUSION: Non-cholera vibrios like V. alginolyticus should be considered as possible causative agents in seawater-contaminated wounds. S. lugdunensis is a relevant pathogen in mixed wound infections. Early microbiological diagnosis and antibiotic-susceptibility testing is crucial to prevent therapeutic failure.