The diagnostic accuracy of serological tests for Lyme borreliosis in Europe: a systematic review and meta-analysis.

BACKGROUND: Interpretation of serological assays in Lyme borreliosis requires an understanding of the clinical indications and the limitations of the currently available tests. We therefore systematically reviewed the accuracy of serological tests for the diagnosis of Lyme borreliosis in Europe. METHODS: We searched EMBASE en MEDLINE and contacted experts. Studies evaluating the diagnostic accuracy of serological assays for Lyme borreliosis in Europe were eligible. Study selection and data-extraction were done by two authors independently. We assessed study quality using the QUADAS-2 checklist. We used a hierarchical summary ROC meta-regression method for the meta-analyses. Potential sources of heterogeneity were test-type, commercial or in-house, Ig-type, antigen type and study quality. These were added as covariates to the model, to assess their effect on test accuracy. RESULTS: Seventy-eight studies evaluating an Enzyme-Linked ImmunoSorbent assay (ELISA) or an immunoblot assay against a reference standard of clinical criteria were included. None of the studies had low risk of bias for all QUADAS-2 domains. Sensitivity was highly heterogeneous, with summary estimates: erythema migrans 50% (95% CI 40% to 61%); neuroborreliosis 77% (95% CI 67% to 85%); acrodermatitis chronica atrophicans 97% (95% CI 94% to 99%); unspecified Lyme borreliosis 73% (95% CI 53% to 87%). Specificity was around 95% in studies with healthy controls, but around 80% in cross-sectional studies. Two-tiered algorithms or antibody indices did not outperform single test approaches. CONCLUSIONS: The observed heterogeneity and risk of bias complicate the extrapolation of our results to clinical practice. The usefulness of the serological tests for Lyme disease depends on the pre-test probability and subsequent predictive values in the setting where the tests are being used. Future diagnostic accuracy studies should be prospectively planned cross-sectional studies, done in settings where the test will be used in practice.

Evaluating frequency, diagnostic quality, and cost of Lyme borreliosis testing in Germany: a retrospective model analysis.

BACKGROUND: Data on the economic impact of Lyme borreliosis (LB) on European health care systems is scarce. This project focused on the epidemiology and costs for laboratory testing in LB patients in Germany. MATERIALS AND METHODS: We performed a sentinel analysis of epidemiological and medicoeconomic data for 2007 and 2008. Data was provided by a German statutory health insurance (DAK) company covering approx. 6.04 million members. In addition, the quality of diagnostic testing for LB in Germany was studied. RESULTS: In 2007 and 2008, the incident diagnosis LB was coded on average for 15,742 out of 6.04 million insured members (0.26%). 20,986 EIAs and 12,558 immunoblots were ordered annually for these patients. For all insured members in the outpatient sector, a total of 174,820 EIAs and 52,280 immunoblots were reimbursed annually to health care providers (cost: 2,600,850€). For Germany, the overall expected cost is estimated at 51,215,105€. However, proficiency testing data questioned test quality and standardization of diagnostic assays used. CONCLUSION: Findings from this study suggest ongoing issues related to care for LB and may help to improve future LB disease management.

Species, developmental stage and infection with microbial pathogens of engorged ticks removed from dogs and questing ticks.

Research into tick-borne diseases implies vector sampling and the detection and identification of microbial pathogens. Ticks were collected simultaneously from dogs that had been exposed to tick bites and by flagging the ground in the area in which the dogs had been exposed. In total, 200 ticks were sampled, of which 104 came from dogs and 96 were collected by flagging. These ticks were subsequently examined for DNA of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Rickettsia spp. and Babesia canis. A mixed sample of adult ticks and nymphs of Ixodes ricinus (Ixodida: Ixodidae) and Haemaphysalis concinna (Ixodida: Ixodidae) was obtained by flagging. Female I. ricinus and adult Dermacentor reticulatus (Ixodida: Ixodidae) ticks dominated the engorged ticks removed from dogs. Rickettsia spp. were detected in 17.0% of the examined ticks, A. phagocytophilum in 3.5%, B. canis in 1.5%, and B. burgdorferi s.l. in 16.0%. Ticks with multiple infections were found only among the flagging sample. The ticks removed from the dogs included 22 infected ticks, whereas the flagging sample included 44 infected ticks. The results showed that the method for collecting ticks influences the species composition of the sample and enables the detection of a different pattern of pathogens. Sampling strategies should be taken into consideration when interpreting studies on tick-borne pathogens.

To test or not to test? Laboratory support for the diagnosis of Lyme borreliosis: a position paper of ESGBOR, the ESCMID study group for Lyme borreliosis.

BACKGROUND: Lyme borreliosis (LB) is a tick-borne infection caused by Borrelia burgdorferi sensu lato. The most frequent clinical manifestations are erythema migrans and Lyme neuroborreliosis. Currently, a large volume of diagnostic testing for LB is reported, whereas the incidence of clinically relevant disease manifestations is low. This indicates overuse of diagnostic testing for LB with implications for patient care and cost-effective health management. AIM: The recommendations provided in this review are intended to support both the clinical diagnosis and initiatives for a more rational use of laboratory testing in patients with clinically suspected LB. SOURCES: This is a narrative review combining various aspects of the clinical and laboratory diagnosis with an educational purpose. The literature search was based on existing systematic reviews, national and international guidelines and supplemented with specific citations. IMPLICATIONS: The main recommendations according to current European case definitions for LB are as follows. Typical erythema migrans should be diagnosed clinically and does not require laboratory testing. The diagnosis of Lyme neuroborreliosis requires laboratory investigation of the spinal fluid including intrathecal antibody production, and the remaining disease manifestations require testing for serum antibodies to B. burgdorferi. Testing individuals with non-specific subjective symptoms is not recommended, because of a low positive predictive value.

Indium 111-monoclonal antimyosin antibody and magnetic resonance imaging in the diagnosis of acute Lyme myopericarditis.

BACKGROUND: Lyme borreliosis is a tick-borne multisystem disorder that may present as self-limiting early or persistent chronic diseases of the skin, nervous system, joints, heart, and other organs. Cardiac involvement has mainly been reported as acute atrioventricular conduction disturbances or transient ventricular dysfunction. METHODS AND RESULTS: We treated a patient with clinical signs of acute myopericarditis and serologic evidence of Lyme borreliosis confirmed by silver staining of endomyocardial biopsy specimens and indium 111-monoclonal antimyosin antibody scan, which we believe has not been reported previously. Additionally, magnetic resonance imaging revealed epicardial and myocardial areas of increased intensity. CONCLUSION: Indium 111-monoclonal antimyosin antibody scanning and magnetic resonance imaging might play an additional important role in assessing and confirming the diagnosis of Lyme carditis in the presence of clinical symptoms and positive serologic findings.

Testing patients with non-specific symptoms for antibodies against Borrelia burgdorferi sensu lato does not provide useful clinical information about their aetiology.

The aim of this study was to determine whether patients with antibodies against Borrelia burgdorferi sensu lato or who report a history of erythema migrans (EM) or tick bite are more likely to have non-specific symptoms such as musculoskeletal pain, fatigue, sensory disorder, and headache. The study group comprised 423 subjects with non-specific symptoms tested for antibodies against B. burgdorferi sensu lato between July 2012 and December 2014 because of suspicion of Lyme borreliosis (LB). Of these, 285 were females (67%) and 138 were males (33%); the median age was 53 years (range, 7-89 years). Patients with a confirmed diagnosis of LB and patients with a known underlying disease that could influence the development of the symptoms were excluded from the evaluation. Subjects were assigned to the seronegative group or to one of three seropositive groups, and the history of EM and tick bite was also recorded. Statistical analysis was performed with single chi-square tests of independence and multiple logistic regression models. No differences in the occurrence of non-specific symptoms were observed between patients grouped according to antibody status. A history of EM showed no significant effect on any of the non-specific symptoms. A history of tick bite was weakly correlated with joint pain and joint swelling (p <0.05). In conclusion, it is highly unlikely that the complaints are related to a previous infection with B. burgdorferi sensu lato. The results show that testing patients with non-specific symptoms for antibodies against B. burgdorferi sensu lato in the everyday clinical setting does not provide any useful information about their aetiology.

Chronic forms of Borrelia burgdorferi infection of the nervous system.

Three European patients had chronic active forms of Borrelia burgdorferi infection of the nervous system, with high titers of antibodies to this spirochete in serum and CSF. Two patients had meningitis for 3 to 4 years, with remissions in one and slowly progressive symptoms in the other. Both had CT lucencies in the basal ganglia. The third patient had lumbosacral plexus neuropathy for 1 year. All three patients responded to intravenous penicillin treatment.

Genetic approaches to cell biology and metabolism of spirochetes.

Genetic analysis and methodology have only comparatively recently been applied to the study of spirochetes. Although genetic transfer procedures for spirochetes are not widely available, there are several examples of progress in genetic analysis of spirochetes by other approaches. Some examples of these approaches are the following. 1) Genes for synthetic pathways in Treponema and Leptospira have been cloned by complementation of Escherichia coli serving as plasmid hosts. 2) The OspA protein of Borrelia burgdorferi has been overexpressed in E. coli without the signal peptide; the recombinant product has been suitable for circular dichroism as well as other biochemical analyses. 3) The heat shock proteins of B. burgdorferi are homologous to heat shock proteins of E. coli. 4) Enzyme activity profiles of B. burgdorferi and other spirochetes show strain heterogeneity and also indicate which biosynthetic and enzymatic activities are conserved within different spirochetes. 5) The gene organization of rRNA genes have revealed differences between spirochetes and other types of bacteria.

Characterization of the first tick isolate of Borrelia burgdorferi from Italy.

We report on the first isolation of a spirochetal organism from Ixodes ricinus ticks of the Trieste area (Northern Italy) which was identified as Borrelia burgdorferi by its reactivity with specific monoclonal antibodies directed against the OSPA and flagella proteins.

In vitro properties of mixtures of fibrin seal and antibiotics.

The incorporation of gentamycin, neomycin and Polymyxin E into fibrin seal results in prolonged clotting time after mixture with 4 NIH-units of thrombin per ml in vitro. The aminoglycoside antibiotics gentamycin and neomycin also diminished fibrin-alpha-chain crosslinkage and, as a consequence, clot rigidity, as demonstrated with clots containing gentamycin. Clotting time and the rate of alpha-chain crosslinking can be adjusted to normal values by the use of higher thrombin concentrations and incorporation of additional factor XIII into the sealing system. Drug release from the clots was similar for all three antibiotics tested and mainly dependent on the concentration gradient between the clot and its environment. Under the conditions of the present study, about 85% of the antibiotic content of fibrin seal clots were released within 72 h.

Detection of Borrelia burgdorferi in patients with Lyme disease by the polymerase chain reaction.

Borrelia burgdorferi, the causative agent of Lyme disease, was detected in patients' serum by DNA amplification using the polymerase chain reaction (PCR). B burgdorferi was pelleted from serum samples by centrifugation (10,000 x g for 10 minutes) and lysed by treatment with ammonium hydroxide (100 degrees C for 15 minutes). Two pairs of "nested" PCR primers complementary to the gene encoding a major outer surface protein (OSP A) of B burgdorferi were used in DNA amplification under standard PCR conditions (Perkin-Elmer Cetus). Two out of five patients with erythema migrans, the characteristic primary skin lesion associated with early Lyme disease, were positive by the PCR. This method could form the basis of a useful routine laboratory test in those cases of early Lyme disease where conventional serological testing commonly yields equivocal or false negative results.

Comparison of SeroMP IgA with four other commercial assays for serodiagnosis of Mycoplasma pneumoniae pneumonia.

Acute phase serum samples from 23 patients with Mycoplasma pneumoniae pneumonia were examined for specific antibodies with microparticle agglutination (MAG)-assay, complement fixation (CF)-test, IgM-specific ELISA, and Immunocard-based IgM-EIA. A novel 3-h IgA-ELISA (SeroMP-IgA) was tested to assess its diagnostic value. For MAG, CF, IgM-ELISA, card-based IgM-EIA, and IgA-ELISA positive results were obtained in 87%, 87%, 91%, 87%, and 100%, respectively. Overall concordance was 78%. Specificity testing of 46 healthy blood donors revealed an optimal diagnostic cut-off range of 22-30 IgA-ELISA binding units (BU)/mL resulting in 91%-100% specificity and 100%-96% sensitivity for serologic diagnosis of acute M. pneumoniae infection.

Specificity of CSF antibodies against components of Borrelia burgdorferi in patients with meningopolyneuritis Garin-Bujadoux-Bannwarth.

The specificity of immunoglobulin (Ig) for components of Borrelia burgdorferi was investigated in cerebrospinal fluid (CSF), serum and in CSF oligoclonal bands from nine patients with meningopolyneuritis Garin-Bujadoux-Bannwarth (MPN-GBB). All patients showed specific IgG and IgM antibodies in serum and CSF when incubated with a lysate of B. burgdorferi strain B 31. Specific antibody was detected in CSF but not in paired serum samples, indicating intrathecal synthesis. Investigation of the specificity of oligoclonal Ig in the CSF revealed oligoclonal bands with specificity for B. burgdorferi in one of the nine patients.

Growth of various bacteria and yeast in a peptide and an element diet.

Solutions of a peptide and an elemental diet were artificially contaminated with several bacterial and yeast strains. Their growth was measured after eight and 24 h incubation at 4 degrees and 22 degrees C. At 4 degrees C no increase in colony count could be observed in either diet within 24 h. At 22 degrees a significant increase of all but the campylobacter strains was obtained after 24 h in the peptide diet. In the elemental diet only the pseudomonas and the yeast strains showed a small increase, the campylobacters were reduced and the colony count of all the other test strains remained constant. As such diets are administered continuously for up to 24 h via a tube into the duodenum for severe intestinal disorders it is suggested (i) that they should be prepared under sterile conditions (ii) that peptide diets should be given for no longer than 8 h and (iii) that solutions of both diets should not be stored for longer than 24 h in the refrigerator.

Bacterial contamination of dialysate in dialysis-associated endotoxaemia.

Bacteriological investigations and endotoxin (ET) determinations were performed during a routine haemodialysis session for six patients. The glucose free dialysate was prepared with untreated tap water. All patients were dialysed for 5 h. Pseudomonas aeruginosa was regularly isolated in numbers up to 10(7) cfu ml-1 from samples of the dialysate inflow, the dialysate site and the dialysate outflow. ET levels in the plasma of the patients increased continuously during haemodialysis and were always higher in the blood outflow line of the dialyzer than in the blood inflow. Despite the high bacterial counts in the dialysate and the increasing ET levels in the patients plasma neither bacteraemia nor fever was observed. The former is due to the impermeability of the dialyzer membrane for bacteria, the latter is explained by low pyrogenicity of P. aeruginosa endotoxin. Inspection of the dialyzer machines revealed that air-traps and heater-unit for the incoming (untreated) tap water before mixing with the dialysate concentrate were the only sites where high bacterial release was feasible, as this part of the machine escaped disinfection due to the construction of these devices. We recommend the regular disinfection of all parts of a dialyzer machine, including heating units, air traps and valves.

Reactivity of neuroborreliosis patients (Lyme disease) to cardiolipin and gangliosides.

A subset of patients (50%) with neuroborreliosis (Lyme disease) showed IgG reactivity to cardiolipin in solid phase ELISA. In addition, a subset of patients with neuroborreliosis (29%) and syphilis (59%) had IgM reactivity to gangliosides with a Gal(beta 1-3) GalNac terminal sequence (GM1, GD1b, and asialo GM1). Anti-ganglioside IgM antibodies were significantly more frequent in these two groups of patients compared to patients with cutaneous and articular Lyme disease, primary antiphospholipid syndrome, systemic lupus erythematosus and normal controls. Correlative evidence and adsorption experiments indicated that antibodies to cardiolipin had separate specificities from those directed against the gangliosides. IgM antibodies to Gal(beta 1-3) GalNac gangliosides appeared to have similar specificities since these were positively correlated and inhibitable by cross adsorption assays. Given the clinical associations of patients with neuroborreliosis and syphilis with IgM reactivity to gangliosides sharing the Gal(beta 1-3) GalNac terminus, we suggest that these antibodies could represent a response to injury in neurological disease or a cross reactive event caused by spirochetes.

Overfeeding macronutrients to critically ill adults: metabolic complications.

Metabolic complications from overfeeding critically ill patients are serious and sometimes fatal. Nutrition care is best provided through repeated evaluation of patients' responses to feeding. Nutrition support may need to be modified over time to maintain metabolic stability and promote recovery. This article describes the etiology of 10 metabolic complications of overfeeding. Guidelines for recommending macronutrients are discussed, as are factors that could increase the risk of overfeeding. Patients who are very small, very large, or very old are particularly vulnerable to overfeeding. Overfeeding protein has led to azotemia, hypertonic dehydration, and metabolic acidosis. Excessive carbohydrate infusion has resulted in hyperglycemia, hypertriglyceridemia, and hepatic steatosis. High-fat infusions have caused hypertriglyceridemia and fat-overload syndrome. Hypercapnia and refeeding syndrome have also been caused by aggressive overfeeding. Dietitians can prevent or curtail the metabolic complications of overfeeding by identifying patients at risk, providing adequate assessment, coordinating interdisciplinary care plans, and delivering timely and appropriate monitoring and intervention. Dietitians need to document complications, interventions, and the outcomes of their clinical care to evaluate the appropriateness of existing nutrition guidelines.

Borrelia burgdorferi, a possible cause of Bell's palsy?

The etiology of Bell's palsy (B.P.) is as yet unknown. In 7 out of 20 patients with B.P. we observed significantly elevated antibodies against Borrelia burgdoferi (IgM and IgG) by means of an indirect Immunofluorescence test (IIFT). Controls were 50 healthy subjects and 25 patients with various neurological disorders. All were of comparable age and sex, with similar residential backgrounds. In this group no significant titer of antibodies against B.burgdorferi were detectable.

Possible causes of symptoms in suspected coronary heart disease but normal angiograms.

BACKGROUND: In patients with suspected coronary heart disease and normal angiography, the causes of cardiac symptoms frequently remain undetermined. A correct diagnosis is desirable, however, since some of the underlying disorders may be curable, treatable, influence prognosis, or induce screening of the relatives. HYPOTHESIS: In such patients, the prevalence of arterial hypertension, hemochromatosis, hypothyroidism, hypoparathyroidism, tachycardiomyopathy, amyloidosis, and neuromuscular disorders as a possible cause for their symptoms and the seroprevalence of micro-organisms, known to cause myocardial damage, were assessed. METHODS: Consecutive patients with normal coronary angiograms were invited for two visits comprising clinical history and investigation, electrocardiograms, blood tests, and echocardiography. Patients were investigated neurologically if unexplained anginal chest pain or creatine kinase elevation persisted or if echocardiography showed isolated left ventricular abnormal trabeculations. RESULTS: In 71 patients (31 women, 40 men, mean age 60 years), the most common cause for cardiac symptoms was hypertension (66%), followed by neuromuscular disorders (13%), tachycardiomyopathy (9%), hypothyroidism (4%), and hemochromatosis (3%). The seroprevalence for Chlamydia species was 90%, Helicobacter pylori 70%, Chlamydia pneumoniae 63%, Borrelia burgdorferi sensu lato 15%, and Rickettsia conorii 10%. No possible cause was found in 24% of the patients. CONCLUSIONS: In patients with suspected coronary heart disease and normal angiograms, hypertension, neuromuscular disorders, tachycardiomyopathy, hypothyroidism, and hemochromatosis should be considered as possible causes.