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Biomedical subjects

S Rauer

Publications and source records attributed to S Rauer.

18 recordsLinked to original sources
PubMed2006

Guidelines on routine cerebrospinal fluid analysis. Report from an EFNS task force.

A great variety of neurological diseases require investigation of cerebrospinal fluid (CSF) to prove the diagnosis or to rule out relevant differential diagnoses. The objectives were to evaluate the theoretical background and provide guidelines for clinical use in routine CSF analysis including total protein, albumin, immunoglobulins, glucose, lactate, cell count, cytological staining, and investigation of infectious CSF. The methods included a Systematic Medline search for the above-mentioned variables and review of appropriate publications by one or more of the task force members. Grading of evidence and recommendations was based on consensus by all task force members. It is recommended that CSF should be analysed immediately after collection. If storage is needed 12 ml of CSF should be partitioned into three to four sterile tubes. Albumin CSF/serum ratio (Qalb) should be preferred to total protein measurement and normal upper limits should be related to patients' age. Elevated Qalb is a non-specific finding but occurs mainly in bacterial, cryptococcal, and tuberculous meningitis, leptomingeal metastases as well as acute and chronic demyelinating polyneuropathies. Pathological decrease of the CSF/serum glucose ratio or increased lactate concentration indicates bacterial or fungal meningitis or leptomeningeal metastases. Intrathecal immunoglobulin G synthesis is best demonstrated by isoelectric focusing followed by specific staining. Cellular morphology (cytological staining) should be evaluated whenever pleocytosis is found or leptomeningeal metastases or pathological bleeding is suspected. Computed tomography-negative intrathecal bleeding should be investigated by bilirubin detection.

Advisory Committees↗
PubMed2006

Antimyelin antibodies and the risk of relapse in patients with a primary demyelinating event.

AIM: To investigate whether the presence of serum antibodies against myelin oligodendrocyte glycoprotein (MOG) and myelin basic protein (MBP) in patients with a clinically isolated syndrome (CIS) predicts the interval to develop more frequently and earlier a first relapse (clinically definite multiple sclerosis: CDMS) than seronegative patients. METHODS: Sera from 45 patients with a CIS and positive intrathecal IgG-synthesis were retrospectively tested for the presence of IgM antibodies against both MOG and MBP. Antibodies were detected by immunoblot using recombinant MOG (1-125) and human MBP antigen preparations. Clinical follow ups were performed retrospectively by telephone interviews and documented neurological examination. RESULTS: Using the Cox proportional hazards model there was no significant increased risk for developing CDMS in anti-MOG and anti-MBP positive patients compared with negative. However regarding the median of the time span between CIS and CDMS over the whole follow up, antibody positive patients (MOG/MBP +/+) developed significantly earlier relapses (median 5.5 months (range 3-20)) than the antibody negative ones (median 25.0 months (range 7-43); p<0.006). On testing sera from 56 apparently healthy students, quite high frequencies of anti-MOG and anti-MBP antibodies (21% and 28% respectively) were detected. This limited specificity of anti-MOG and anti-MBP antibodies has been seen earlier and restricts their diagnostic relevance in MS despite their role as a predictor of relapses after a CIS. CONCLUSIONS: This study confirms previous data only in a subanalysis indicating that patients with positive anti-MOG/MBP antibodies develop earlier relapses than patients who are antibody negative. However, the authors could not verify that the presence of these antibodies anticipates the overall risk of developing CDMS-according to study criteria-after a first demyelinating event within the study period of 21-106 months (mean 60 (SD 25)).

Adolescent↗
PubMed2005

Fulminant acute disseminated encephalomyelitis mimicking acute bacterial menigoencephalitis.

Most patients with acute disseminated encephalomyelitis (ADEM) recover quickly under corticosteroid treatment and have a favourable long-term prognosis. We report on a young woman with acute onset of an extensive and solitary white-matter lesion in the left hemisphere. Fever, high pleocytosis and elevated protein in cerebrospinal fluid initially suggested bacterial meningoencephalitis. The patient died from brain herniation despite maximal conservative therapy. Histological changes in necropsy were consistent with the diagnosis ADEM. Treatment options of fulminant ADEM are discussed.

Adult↗
PubMed2001

Avidity determination of Borrelia burgdorferi-specific IgG antibodies in Lyme disease.

The avidity indices of Borrelia burgdorferi-specific IgG antibodies were estimated using ELISA in sera from patients with different stages of Lyme disease. In addition, sera from healthy students with proof of borrelial-specific IgG antibodies from standard serology were tested. Low avidity indices were detected predominantly in sera from patients with early-stage Lyme disease [erythema migrans (EM); n = 25]. High avidity indices were found in healthy students (n = 72) and in most of the patients with neuroborreliosis (NB; n = 44) and chronic late-stage Lyme disease [acrodermatitis chronica atrophicans (ACA); n = 36]. In conclusion, early-stage Lyme disease (EM) could be differentiated from advanced and chronic stages (NB, ACA) and from "seropositive" healthy persons using avidity determination in the majority of patients in this study.

Antibody Affinity↗
PubMed2001

Enzyme linked immunosorbent assay using recombinant HuD-autoantigen for serodiagnosis of paraneoplastic neurological syndromes.

OBJECTIVES: The aim of this study was to evaluate the suitability of an ELISA employing purified recombinant HuD antigen, for detection of specific anti-HuD antibodies in sera and cerebrospinal fluid (CSF) from patients with paraneoplastic neurological syndromes (PNS). MATERIAL AND METHODS: The cutoff for optical density readings was estimated by testing of 145 sera from healthy subjects. Sera from 17 patients with paraneoplastic neurological syndromes (PNS) and evidence of a clear anti-Hu band pattern in an immunoblot employing human cerebellar crude extract as antigen were tested. RESULTS: All 17 sera from patients with PNS revealed a clear positive result in the HuD-ELISA, demonstrating a sensitivity of 100%. Two out of 150 sera from patients with various infectious, autoimmune and neoplastic diseases (excluding PNS) showed a borderline result in the HuD-ELISA. This reveals a specificity of more than 98%. In addition 10 serum/CSF pairs from patients with anti-Hu-syndrome were adjusted to equal IgG concentrations and were tested in parallel in the HuD-ELISA. A specific antibody index (AI=ODCSF/ODserum) over 1.5 indicates intrathecal antibody synthesis. Six of ten patients revealed an AI >1.5, one was borderline (AI=1.5), and three had an AI in the range of 0.9-1.2. There appears to be a correlation between elevated AIs (>1.5) and presence of oligoclonal bands in the CSF. CONCLUSION: Due to its high specificity and sensitivity the recombinant HuD-ELISA is a suitable test for detection of anti-HuD antibodies in patients with putative PNS. In addition, the HuD-ELISA seems to be appropriate for the detection of specific intrathecal HuD-antibody synthesis.

Adult↗
PubMed2000

Demonstration of anti-HuD specific oligoclonal bands in the cerebrospinal fluid from patients with paraneoplastic neurological syndromes. Qualitative evidence of anti-HuD specific IgG-synthesis in the central nervous system.

The presence of HuD-specific oligoclonal IgG bands in the CSF was investigated in five patients with paraneoplastic neurological syndromes. All patients revealed intrathecal synthesis of HuD specific antibodies in the CSF, as estimated from elevated antibody indices (>1.5) in an IgG-ELISA using recombinant HuD-protein as antigen. Isoelectrofocussing combined with affinity blotting showed reactivity of IgG bands with recombinant HuD antigen in all CSF samples. These data support the idea that HuD specific antibodies in the CSF are produced mainly by B-cell clones in the central nervous system. These findings support the hypothesis of autoimmunity in the pathogenesis of anti-Hu associated paraneoplastic neurological syndromes.

Aged↗
PubMed1999

Advantage of recombinant borrelial proteins for serodiagnosis of neuroborreliosis.

Two enzyme immunoassay (EIA) systems were compared for their ability to detect Borrelia burgdorferi sensu lato specific IgG and IgM antibodies and to differentiate between symptomatic (83 patients with neuroborreliosis) and asymptomatic seropositive subjects (80 healthy controls). Antibody concentrations were determined by EIA; the antigens used were either a sonicate of B. burgdorferi or three recombinant borrelial proteins: the 14-kDa flagellin fragment, the outer surface protein C (22 kDa) and the high molecular mass protein p83 (83 kDa). In the sonicate, EIA, IgG or IgM antibodies to B. burgdorferi, or both, were detected in all patients with neuroborreliosis and in all controls. Pre-absorption of sera with Treponema phagedenis sonicate diminished the sensitivity of detection of borrelial specific IgG (IgG or IgM or both) antibodies in patients with neuroborreliosis from 80 to 57% (100 to 82%) and in the controls from 100 to 32% (100 to 37%). While being specific for B. burgdorferi, the recombinant EIAs proved to be significantly more sensitive than the sonicate EIA: IgG or IgM, or both antibodies against any of the recombinant antigens were detected in 92% of patients with neuroborreliosis and in 24% of controls. The increase in sensitivity in patients with neuroborreliosis was mostly due to the higher detection rate of IgM antibodies in the recombinant EIA (77% versus 48% in the sonicate EIA), while IgG antibodies were demonstrated with similar frequencies in both EIA systems (57% versus 60%). It was concluded that the recombinant EIAs are superior to the sonicate EIA with pre-absorption of cross-reactive antibodies in the confirmation of an acute borrelial infection and in the differentiation between symptomatic and asymptomatic infections.

Acute Disease↗
PubMed1998

Analysis of the intrathecal immune response in neuroborreliosis to a sonicate antigen and three recombinant antigens of Borrelia burgdorferi sensu stricto.

The intrathecal synthesis of borrelial-specific IgM- and IgG-antibodies was studied in 67 patients with neuroborreliosis and in 14 patients with neurosyphilis (controls). Antibody concentrations in serum and in the cerebrospinal fluid were determined by an enzyme immunoassay (EIA) using, as antigens, a sonicate of Borrelia burgdorferi, the recombinant 14 kDa flagellin fragment, the outer surface protein C (22 kDa), and the high molecular mass protein p83 (83 kDa). In the sonicate EIA, IgG- and/or IgM-antibodies to Borrelia burgdorferi in serum were detected in all patients with neuroborreliosis and in 71% of patients with neurosyphilis. Intrathecal synthesis of borrelial-specific IgG- and/or IgM-antibodies was demonstrated in 82% of patients with neuroborreliosis and in 71% of patients with neurosyphilis. Immunoglobulin G- and/or IgM-antibodies in serum against any of the recombinant antigens were detected in 92% of patients with neuroborreliosis and in none of those with neurosyphilis. Intrathecal synthesis of IgG- and/or IgM-antibodies to individual recombinant antigens was demonstrated in 67% of patients with neuroborreliosis and in none of those with neurosyphilis. The sensitivity of the recombinant antigens in serum was almost equal to that of the sonicate EIA, whereas the recombinant antigens were clearly less sensitive in the estimation of the intrathecal specific immune response. It was concluded that in suspected cases of neuroborreliosis, the estimation of high specific antibodies in the recombinant EIA will be helpful in confirming the diagnosis.

Antibodies, Bacterial↗
PubMed1998

Enzyme-linked immunosorbent assay using recombinant OspC and the internal 14-kDa flagellin fragment for serodiagnosis of early Lyme disease.

The outer surface protein C (OspC) and the internal 14-kDa flagellin fragment of strain GeHo of Borrelia burgdorferi sensu stricto were expressed as recombinant proteins in Escherichia coli and were purified for use in an immunoglobulin M (IgM) enzyme-linked immunosorbent assay (OspC-14-kDa antigen ELISA). No hint at disturbing protein-protein interferences, which might influence the availability of immunoreactive epitopes, was found when the recombinant antigens were combined in the ELISA. The recombinant OspC-14-kDa antigen ELISA was compared to a commercial IgM ELISA that used a detergent cell extract from Borrelia afzelii PKo as the antigen. According to the manufacturer's information, the cell extract contains, in addition to other antigens, the following diagnostically relevant antigens: the 100-kDa (synonyms, 93- and 83-kDa antigens), 41-kDa, OspA, OspC, and 17-kDa antigens. The specificity was adjusted to 95% on the basis of data for 154 healthy controls. On testing of 104 serum samples from patients with erythema migrans (EM), the sensitivity of the recombinant ELISA (46%) for IgM antibodies was similar to that of the commercial ELISA (45%). However, when 42 serum samples from patients with polyclonal B-cell stimulation due to an Epstein-Barr virus infection were tested, false-positive reactions were significantly less frequent in the recombinant ELISA (10%) than in the whole-cell-extract ELISA (23%). OspC displays sequence heterogeneity of up to 40% according to the genomospecies. However, when the reactions of serum specimens from controls and EM patients with OspC from representative strains of B. burgdorferi sensu stricto (strain GeHo) and B. afzelii (strain PKo) were compared in an ELISA, almost no differences in specificity and sensitivity were seen. This demonstrates that the sera predominantly recognize the common epitopes of OspC tested in this study. In conclusion, we suggest that the OspC-14-kDa antigens ELISA is a suitable test for the detection of an IgM response in early Lyme disease.

Antibodies, Bacterial↗
PubMed1997

[Follow-up and prognosis of early summer meningoencephalitis].

Sixty-three patients with tick-borne encephalitis were studied for sequelae up to 5 years after the acute illness (median: 12 months, range: 1-44 months). Patients were examined clinically, by neuropsychological testing and by electroencephalography. The clinical presentation during the acute stage was as follows: Meningitis (M,n = 12), Meningoencephalitis (Me,n = 27), Meningoencephalomyelitis (My,n = 15), and Meningoencephaloradiculitis (R,n = 9). A total of 59 patients reported a neurasthenic syndrome after discharge, which correlated with the severity of the acute illness. Twenty patients were not able to work because of reduced stress tolerance, fatigue or an elevated emotional sensitivity, which lasted for 3 months at most. In some patients hypacusis (n = 7), severe dysarthria and dysphagia (n = 4) remained essentially unimproved for years following the acute illness. While in 8/9 patients with radiculitis paresis of the extremities improved well over months to years, improvement was quite limited in all patients with myelitis. In 41/55 patients, investigations by electroencephalography revealed normal findings even within months after acute illness. Persistent cognitive deficits were present only in 7/11 patients with a severe course of disease.

Adolescent↗
PubMed1995

Establishment of enzyme-linked immunosorbent assay using purified recombinant 83-kilodalton antigen of Borrelia burgdorferi sensu stricto and Borrelia afzelii for serodiagnosis of Lyme disease.

The 83-kDa antigen of Borrelia burgdorferi was expressed as a recombinant protein in Escherichia coli and purified for use in an enzyme-linked immunosorbent assay (p83-ELISA). Antibodies to the 83-kDa antigen of both the immunoglobulin G (IgG) and IgM isotypes could be detected in all stages of Lyme disease. Sensitivity varied, depending on the clinical stage of illness. In early stages, as defined for 118 patients with erythema migrans, it was found to be 20% (24 of 118 patients: 7 with IgM, 16 with IgG, and 1 with IgM and IgG). Of the patients with late-stage Lyme arthritis and acrodermatitis chronica atrophicans, 94% (16 of 17:2 with IgM and IgG and 14 with IgG) and 86% (36 of 42:2 with IgG and IgM and 34 with IgG) revealed positive results in the p83-ELISA, respectively. p83 displays sequence heterogeneity according to the genomospecies, but when the reactions of serum specimens from acrodermatitis chronica atrophicans patients and arthritis patients with p83 derived from representative strains of B. burgdorferi sensu stricto and Borrelia afzelii in ELISAs were compared, no differences in specificity and sensitivity were seen. When 82 serum specimens from healthy controls were tested, none had IgG and only 3 (4%) had IgM antibodies, indicating a high specificity. Positive reactions with antibodies against Treponema pallidum (1 of 37 patients; IgG) and Epstein-Barr virus (1 of 44 patients; IgM) and with autoantibodies of various specificities (1 of 53 patients; IgG) were seen with < 3% of the serum samples te11111111111111111111 high speficicity for B. burgdorferi.2+ 13% for IgM antibodies, the IgM p83-ELISA provided little diagnostic information for Lyme disease, whereas the IgG p83-ELISA appears to be a suita ;e test for serodiagnosis of advanced-stage Lyme disease.

Antibodies, Bacterial↗
PubMed1994

Use of a hybrid protein consisting of the variable region of the Borrelia burgdorferi flagellin and part of the 83-kDa protein as antigen for serodiagnosis of Lyme disease.

A hybrid protein consisting of the variable region of the Borrelia burgdorferi flagellin (an 18-kDa fragment) and a 59-kDa fragment (lacking the N-terminal part) of the 83-kDa protein has been constructed by genetic engineering. It was expressed as a nonfusion protein of an apparent molecular weight of 77,000 in Escherichia coli. The suitability of this new antigen for the diagnosis of Lyme disease was tested by immunoblotting; for comparison, the recombinant variable region of the flagellin, the 18-kDa fragment (p18), and the whole recombinant 83-kDa protein (p83), both expressed in E. coli, were used. A total of 120 serum samples from various stages of Lyme disease, which were positive in two serological assays, a passive hemagglutination assay and an indirect immunofluorescence assay, were tested. By indirect immunofluorescence, 74 samples were positive for immunoglobulin G (IgG) antibodies and 72 were positive for IgM antibodies. Of these serum samples, 69 of 74 (93%) contained IgG antibodies against p18 and/or p83, and IgG antibodies were detected by the hybrid protein in 67 (90%) samples. IgM antibodies against p18 and/or p83 were detected in 60 of 72 (83%) serum samples, and 57 (79%) serum samples were reactive with the hybrid protein. Twenty serum samples of patients with a history of syphilis and 40 serum samples, negative in routine B. burgdorferi serology, were tested as controls. The hybrid protein, made up of specific epitopes of an early (p18) and late (p83) antigen, is recognized by almost the same number of patient serum samples as the individual antigens.

Antigens, Bacterial↗
PubMedDate not supplied

Serodiagnosis of neuroborreliosis: comparison of reliability of three confirmatory assays.

The sensitivity and specificity of three confirmatory assays for the serodiagnosis of neuroborreliosis were investigated. Samples from 96 patients with proven neuroborreliosis, 80 healthy volunteers, 20 patients with neurosyphilis and 20 patients with recent infections with Epstein-Barr virus (EBV) were tested for borrelial antibodies by immunoblotting, Borrelia burgdorferi sensu lato sonicate EIA following pre-absorption of cross-reactive antibodies (Abs-EIA) and by a so-called RECO-EIA using the following recombinant borrelial proteins as antigens: a 14 kDa-internal flagellin fragment, the outer surface protein C (23 kDa) and the high molecular mass protein p83 (83 kDa). The immunoblots were evaluated according to the criteria published by Engström et al. and Hauser et al. An evaluation of IgM and/or IgG antibodies revealed a considerably higher sensitivity for the RECO-EIA (94%) compared to the Abs-EIA (82%, P < 0.0001). Evaluation of the immunoblot according to the criteria of Hauser was significantly more sensitive than according to the criteria of Engström (89 vs 51%, P = 0.0003). A higher sensitivity was demonstrated for IgM (54 vs 22%) and IgG antibodies (64 vs 24%). When both findings from RECO-EIA and immunoblotting were considered, positive findings in the first step assay (sonicate EIA without pre-absorption) were confirmed in 97% of patients. When samples were tested for IgM antibodies, the specificities of the three confirmatory assays did not differ significantly, but in the case of IgG antibodies, the immunoblot (Hauser: P = 0.013; Engström: P = 0.004) and the RECO-EIA (P = 0.02) were more specific than the Abs-EIA. It is concluded that the immunoblot (evaluated according to Hauser) and the RECO-EIA are both suitable as confirmatory assays in the serological diagnosis of neuroborreliosis. Monoclonal antibodies are mandatory tools in the evaluation of the immunoblot.

Absorption↗