Detection Of Oligoclonal IgG Bands Research Articles

IgM to phosphatidylcholine in multiple sclerosis patients: from the diagnosis to the treatment.

Multiple sclerosis (MS) is a demyelinating and neurodegenerative disease of the central nervous system. It affects young people, and a considerable percentage of patients need the help of a wheelchair in 15 years of evolution. Currently, there is not a specific technique for the diagnosis of MS. The detection of oligoclonal IgG bands (OIgGBs) is the most sensitive assay for it, but it is not standardizable, only reference laboratories develop it, and uses cerebrospinal fluid. To obtain this sample, a lumbar puncture is necessary, an invasive proceeding with important side effects. It is important to develop and implement standard assays to obtain a rapid diagnosis because the earlier the treatment, the better the evolution of the disease. There are numerous modifying disease therapies, which delay the progression of the disease, but they have important side effects, and a considerable percentage of patients give up the treatment. In addition, around 40% of MS patients do not respond to the therapy and the disease progresses. Numerous researches have been focused on the characterization of predictive biomarkers of response to treatment, in order to help physicians to decide when to change to a second-line treatment, and then the best therapeutic option. Here, we review the new biomarkers for the diagnosis and response to treatment in MS. We draw attention in a new assay, the detection of serum IgM to phosphatidylcholine, that showed a similar sensitivity as OIgGBs and predicts the response to disease modifying treatments.

Investigation of Oligoclonal IgG Bands in Tear Fluid of Multiple Sclerosis Patients.

Background: Oligoclonal IgG bands (OCB) in the cerebrospinal fluid (CSF) represent a typical marker for inflammation in multiple sclerosis (MS) patients and have a predictive and diagnostic value in patients with a first suspected demyelinating event. The detection in tears remains controversial but some reports suggested a replacement of CSF analysis by OCB detection in tears. We aimed to investigate the value of OCB detection in tears systematically in patients with MS.Methods: Tears of 59 patients with suspected or diagnosed MS were collected with Schirmer filter paper strips. Tear IgG was purified by affinity chromatography with protein G. After isoelectric focusing in polyacrylamide gels OCB detection was performed with direct silver staining. Paired triplets of CSF, serum, and tears were analyzed. For comparison purposes we additionally used other tear collection methods (flush procedure and plastic capillary tubes) or detection techniques (Immunoblotting). Clinical and paraclinical parameters are provided.Results: IgG collection in tears was most reliable by using Schirmer strips. Thirteen patients had to be excluded due to insufficient sample material. Tear specific proteins that interfered with OCB detection were successfully eliminated by IgG purification. The concordance of OCB in tears and CSF of all investigated MS patients was 39% with a high rate of only marginal pattern in tears. Five patients demonstrated restricted bands in tears, neither detectable in CSF nor serum. Occurrence of OCB in tears was significantly associated with pathological visual evoked potentials (P = 0.0094) and a history of optic neuritis (P = 0.0258).Conclusion: Due to the limited concordance, high rate of samples with insufficient material, and the unknown origin of tear IgG we cannot recommend that tear OCB detection may replace CSF OCB detection in MS patients. The detection of unique OCB in tears might offer new insights in ophthalmological diseases.

BAFF is decreased in the cerebrospinal fluid of multiple sclerosis at clinical onset

B-cells are thought to play a relevant role in multiple sclerosis (MS) pathology. BAFF (B cell activating factor of the TNF family) is a B-cell survival factor constitutively produced inside the CNS by astrocytes. We studied the intrathecal synthesis of BAFF in MS at clinical onset.Paired serum and cerebrospinal fluid (CSF) specimens from 40 clinically isolated syndromes (CIS) suggestive of MS or early relapse-onset MS (eRRMS) and from 18 healthy controls (HC) were analysed. Patients were classified based on the detection of oligoclonal IgG bands in the CSF (IgGOB+ and IgGOB−). BAFF was detected by highly sensitive ELISA and its ratio (CSF-BAFF/serum-BAFF, QBAFF) and Index (QBAFF/QAlb, BAFF-Index) were calculated.IgGOB+ presented lower CSF concentrations of BAFF compared to both HC and IgGOB− (p<0.05). BAFF Index was significantly lower in IgGOB+ compared to both HC and IgGOB− (p<0.01). A significant inverse correlation between QIgG and QBAFF (r: −0.4, p<0.05) and between BAFF index and IgGIF (r: −0.4, p<0.05) or IgG Index (r: −0.4, p=0.05) was found in IgGOB+. The decreased CSF levels of BAFF in IgGOB+ at clinical onset suggest the absorption of this factor by intrathecally recruited B cells since the early disease phases.

K Index in cerebrospinal fluid: a valid tool in multiple sclerosis diagnosis

Detection of oligoclonal IgG bands in cerebrospinal fluid by isoelectrocfocusing and immunodetection is the current gold standard to detect an inflammatory process in the central nervous system. It has been proposed that the presence of free light chains (FLCs) in CSF was associated with recent demyelination activity in MS and might be used as a prognosis marker. Our study’s objective is assessing the diagnostic accuracy of a new highly sensitive latex-enhanced nephelometric assay for k free light chain (kFLC) determination in CSF/serum as an alternative to traditional tests and its clinical application. kFLCs were measured in CSF/serum pairs from 80 patients by the use of a new highly sensitive latex-enhanced nephelometric automated immunoassay for detection of immunoglobulin FLC. The eighty patients were split into three groups according to the neurological diagnosis. In this study we confirm even more the use of the k Index as a diagnostic aid in multiple sclerosis. kFLC Index seems to be more accurate parameter respect the determination of oligoclonal immunoglobulin bands (OCBs). We recalculate the K Index sensitivity and specificity respect the precedent published result. Two patients previously diagnosed with leukoencephalopathy have gone to group 3 as confirmed the diagnosis of MS. These new data reinforce even more the use of the k Index to diagnose MS in comparison to classical methods and to the reference method, the OCBs.

Optimization of supervised self-organizing maps with genetic algorithms for classification electrophoretic profiles

&lt;p&gt;Standard electrophoresis methods were used in the classification of analyzed proteins in cerebrospinal fluid from patients with multiple sclerosis. Disc electrophoresis was carried out for detection of oligoclonal IgG bands in cerebrospinal fluid on polyacrylamide gel, mainly with multiple sclerosis and other central nervous system dysfunctions. ImageMaster 1D Elite and GelPro specialized software packages were used for fast accurate image and gel analysis. The classification model was based on supervised self-organizing maps. In order to perform the modeling in automated manner genetic algorithms were used. Using this approach and a data set composed of 69 samples we were able to develop models based on supervised self-organizing maps which were able to correctly classify 83 % of the samples in the data set used for external validation.&lt;/p&gt;

The utility of cerebrospinal fluid analysis in patients with multiple sclerosis

Diagnosis of multiple sclerosis (MS) requires the exclusion of other possible diagnoses. For this reason, the cerebrospinal fluid (CSF) should be routinely analysed in patients with a first clinical event suggestive of MS. CSF analysis is no longer mandatory for diagnosis of relapsing-remitting MS, as long as MRI diagnostic criteria are fulfilled. However, caution is required in diagnosing MS in patients with negative MRI findings or in the absence of CSF analysis, as CSF investigation is useful to eliminate other causes of disease. The detection of oligoclonal IgG bands in CSF has potential prognostic value and is helpful for clinical decision-making. In addition, CSF analysis is important for research into the pathogenesis of MS. Pathophysiological and neurodegenerative findings of inflammation in MS have been derived from CSF investigations. Novel CSF biomarkers, though not yet validated, have been identified for diagnosis of MS and for ascertaining disease activity, prognosis and response to treatment, and are likely to increase in number with modern detection techniques. In this Review, we summarize CSF findings that shed light on the differential diagnosis of MS, and highlight the potential of novel biomarkers for this disease that could advance understanding of its pathophysiology.

Cluster analysis method in electrophoretic patterns classification

The conventional electrophoresis methods are well known techniques for protein detection and analysis of cerebrospinal fluid (CSF). Disc electrophoresis (DEP) was carried out for detection of oligoclonal IgG bands in cerebrospinal fluid (CSF) on polyacrilamide gel. However, the advance of automation has made rapid collection of large amounts of data feasible and the development of microcomputers has made sophisticated processing even of old electrophoregrams possible. Automated analysis, data storage and sophisticate data acquisition were carried out with Gel Pro Analyzer 3.1, which is specifically structured to analyze gels and elctrophoregrams: complex band pattern matching (gel variation, dendogram analysis etc.); lane relation studies (sophisticated lane database); general gel analysis (accurate molecular size, quantitative determination of protein mixture etc.). Clustering techniques have been applied for detection of intrathecal immune response. Different hierarchic cluster analysis methods such as single linkage, complete linkage, unweighted pair-group average (UPMGA) were used. In addition, other cluster characteristics such, distance matrix and Euclidean distance matrix were calculated. Pairing of electrophoresis methods and cluster image analysis, could lead to additional diagnostic information of inflammatory conditions of the central nervous system (CNS) or dysfunction of blood-CSF barrier.

Place de l'immunofixation des IgG du LCR dans le diagnostic de sclérose en plaques (SEP)

We presented the analytical and diagnostic performances of immunofixation for detection of oligoclonal IgG bands in native unconcentrated cerebrospinal fluid (CSF) and diluted serum samples analysed in the same run on Hydragel 6 CSF Sebia on Hydrasys ® Sebia, an alternative to the manual technique of IgG isoelectric focusing (IEF). Pairs of CSF and serum of 124 patients studied over a 2 year period were classified in: 29 definite multiple sclerosis (MS) 35 possible MS, 11 inflammatory neurological diseases other than MS, and 49 non-inflammatory diseases of the central nervous system. Pairs of CSF and serum of 98 patients were tested by isoelectric focusing for control of intrathecal IgG synthesis. The analytical performances of the two techniques to reveal IgG oligoclonal bands are practically identical. The sensivity of immunofixation is 96%, of isoelectric focusing is 93%. Their specificity is 93% and 96%, respectively. The positive predictive value to establish diagnosis of MS is 90% with immunofixation and 92% with isoelectric focusing. The negative predictive value to exclude diagnosis is 97% with immunofixation and 96% with isoelectric focusing. The interpretation of IgG immunofixation is in practice more difficult than IgG isoelectric focusing, the procedure is easy and quick to perform.

An ultrasensitive method for the detection of oligoclonal IgG bands.

We have developed an ultrasensitive isoelectrofocusing (IEF) method for the detection of oligoclonal (OGC) IgG bands in cerebrospinal fluid (CSF) and sera. In this procedure, double antibody and peroxidase immunodetection have been substituted by a single antibody labelled with alkaline phosphatase. Alkaline phosphatase immunodetection not only improves tenfold the sensitivity of the assay but also gives a sharper pattern resolution making the interpretation easier and increases the specificity of this technique. This preliminary report should be validated in a larger number of patients with and without multiple sclerosis (MS).

Clinical characteristics and neurophysiologic findings in patients with multiple sclerosis without oligoclonal IgG in cerebrospinal fluid

Besides magnetic resonance imaging, the presence of locally produced oligoclonal IgG bands (OCB) in the cerebrospinal fluid (CSF) is the most consistent laboratory abnormality in patients with multiple sclerosis (MS). The most sensitive method for the detection of CSF OCB is isoelectric focusing (IEF) [6]. Occasional patients with clinically definite MS lack evidence for intrathecal IgG synthesis [7, 8]. This study was designed to compare clinical data and evoked potential (EP) findings between CSF OCB positive and OCB negative MS patients. The study comprised 22 OCB negative patients with clinically definite MS [11] and 22 OCB positive controls matched for age, disease duration, activity and course of MS. In both groups clinical assessment was performed by using Expanded Disability Status Scale (EDSS) score [12] and progression rate (PR). All patients underwent multimodal EP: visual (VEPs), brainstem auditory (BAEPs) and median somatosensory (mSEPs). The VEPa were considered abnormal if the P100 latency exceeded 117 ms or inter-ocular difference greater than 8 ms was detected. The BAEPs were considered abnormal if waves III or V were absent or the interpeak latencies I-III, III-V, or I-V were increased. The mSEPs were considered abnormal when N9, N13 and N20 potentials were absent or when increased interpeak latencies were recorded. The severity of the neurophysiological abnormalities was scored for each modality as follows: normal EP score 0; every other EP abnormality except the absence of one of the main waves, score 1; absence of one or more of the main waves, score 2 [13]. Both mean EDSS score (4.0 vs. 3.5) and PR (0.6 vs. 0.5) were similar in OCB positive and OCB negative group, (p > 0.05). In the first group males were predominant, but without statistical significance (Table 1). Disease started more often with the brainstem symptoms in the OCB positive than in OCB negative MS group (p = 0.028), while there was no differences in other initial symptoms between the groups (Graph 2). The frequency of (multimodal) EP abnormalities was higher in the OCB positive group but the differences were not statistically significant, except for bilateral SEP abnormalities (p = 0.012). The severity of the AEPs abnormalities was similar in both groups while for the VEPs and SEPs abnormalities were more pronounced in the OCB positive group but not significantly (Table 2). The male preponderance of OCB negative MS patients in our study is in accordance with previous studies [14, 15]. This finding could be potentially ascribed to the well known gender-related differences in both humoral and cellular immune responses [17]. We found no statistically significant differences in either disability or PR between the two patient groups, although OCB negative MS patients had lower EDSS score and PR than OCB positive cases. In accordance with these findings, Fukazawa et al. also failed to show differences in disability between OCB negative and positive MS patients. On the other hand, few studies reported that OCB negative MS patients have a better prognosis [16, 18]. The only clinical difference between two groups of patients that we found was that the disease more often started with brainstem symptoms in OCB positive MS patients (p = 0.028). OCB positive MS patients had more often bilateral SEPs abnormalities (p = 0.012). There was no statistically significant differences between two groups of patients in the severity of trimodal EPs abnormalities and the frequency of BAEPs and VEPs abnormalities although OCB negative patients had trend towards less pronounced EP disturbancies. Our results did not reveal significant difference in clinical and neurophysiological(y) parameters between two groups of patients. However, they indicate a trend towards better prognosis of the disease in OCB negative MS patients.

Magnetic resonance findings in the brain of patients with multiple sclerosis without oligoclonal bands in the cerebrospinal fluid

Locally produced oligoclonal IgG bands (OCB) are present in the cerebrospinal fluid (CSF) of 95% patients with multiple sclerosis (MS) [2,3]. The most sensitive method for the detection of OCB is isoelectric focusing (IEF) [1]. Occasional patients with clinically definite MS lack evidence for intrathecal IgG synthesis [2,9]. This study was designed to compare brain magnetic resonance imaging (MRI) findings between CSF OCB positive and negative MS patients. The study comprised 22 OB negative patients with clinically definite MS and 22 OCB positive controls matched for age, disease duration, activity and course of MS. In the both groups clinical assessment was performed by using Expanded Disability Status Scale (EDSS) score. T2 weighted MRI of the brain was performed on a Siemens Magnetom (1.0T). Lesions were counted and sized for 15 anatomically defined locations: 7 periventricular (PV) and 8 non-periventricular (NPV) regions. An arbitrary scoring system weighted for lesions size was used to estimate total and regional lesions loads: a) 1 point was given for each lesion with a diameter 1-5 mm, b) 2 points for one lesion with a diameter 6-10 mm, c) 3 points for one over 10 mm, and confluent lesions scored one extra point [16]. Atrophy were scored as follows: 0-normal size, 1-mild atrophy, 2-moderate atrophy and 3-severe atrophy. Mean score of total brain MRI loads was lower in OCB negative than in OCB positive MS patients (44 vs. 50) but the difference was not statistically significant. Mean periventricular (32 vs. 23), non-periventricular (26 vs. 19) and infratentorial (11 vs. 9) scores were higher in OCB positive MS group in comparison with OCB negative patients, but non-significant (figure 1). There was no correlation between EDSS score and total MRI lesions load in OCB negative MS patients, while in OCB positive group we detected significant correlation between EDSS score and total MRI lesions load (p = 0.026) (figure 2). The results of this study demonstrate that by using conventional brain MRI the extent end severity of the pathological process seems to be similar in OCB negative and OCB positive MS patients. On the other hand, we found statistically significant correlation between brain MRI total lesion load and EDSS in the OCB positive MS patients, while this correlation was not detected in OCB negative MS patients. Differences in brain MRI findings between OCB positive nad OCB negative MS patients have been already analyzed [9,12]. In the first, Zeman et al. reported that OCB negative MS patients have lower total MRI brain lesion loads in comparison to OCB positive group, but the differences was not statistically significant [9]. In accordance with these findings, Fukazawa et al. also failed to show differences in the distribution, extent, shape and number of brain MRI lesions between OCB positive and negative MS patients. [12]. On the other hand, it has been demonstrated that the rate of intrathecal IgG synthesis apparently correlates with plaque volume in the brain, as demonstrated on MRI, in MS patients [17]. However, our results along with those from two above-mentioned previous studies do not support this notion. In conclusion, trend towards lesser MRI lesion load and lack of its correlation with EDSS in OCB negative MS patients, warrants further investigations with new MRI techniques (magnetic resonance spectroscopy and magnetisation transfer), including the thorough exploration of normal-appearing while matter, in OCB negative MS patients.

Comparison of Methods for the Detection of Oligoclonal IgG Bands in Cerebrospinal Fluid and Serum: Results of the Dutch Quality Control Survey

Detection of oligoclonal IgG bands (OCBs) in parallel cerebrospinal fluid (CSF) and serum samples is an important tool to support the diagnosis of neuroinflammatory diseases such as multiple sclerosis (MS) (1). Because ∼90–95% of the patients diagnosed with MS present with OCBs in the CSF (2), whereas other CSF indices may be normal, this type of analysis has long been recommended for the diagnosis of MS, especially when the clinical presentation does not clearly suggest this diagnosis (1)(3). CSF-restricted OCBs are also essential for the diagnosis of primary progressive MS (1). Furthermore, in several neuroimmunologic disorders, including neuroborreliosis, neurosyphilis, and viral infections of the brain, the identification of OCBs in CSF may be helpful in the diagnostic process (2)(4)(5). Twice a year, pairs of CSF and serum samples are distributed by our laboratories to the participants of the Dutch CSF Quality Control survey for analysis of OCBs. It was the aim of this study to evaluate the performance of the 56 participating laboratories for OCB analysis with regard to both the methods used for protein separation and detection of OCBs and to derive a general recommendation on laboratory management for this type of analysis. To collect a sufficient volume of CSF, samples stored at −80 °C and selected for the absence of OCBs were pooled. Sera taken from healthy controls were collected and pooled as well. Sera containing either a known OCB pattern or monoclonal IgG were also collected with informed consent from patients. We added minimal amounts of these clinical serum samples to the CSF pool to create artificial patient CSF samples. In each survey, an OCB-enriched CSF sample and a serum sample were sent out as a pair of clinical CSF and serum samples. In this way we were able to …

Respuesta inmune intracraneal oligoclonal y poliespecífica en la esclerosis múltiple

The intrathecal response of IgG, is the most frequent neuropathological sign in multiple sclerosis (MS) patients beside the detection of oligoclonal IgG bands in cerebrospinal fluid. At the same time the observation of an intrathecal antibody synthesis (antibody index> 1.4) against neurotropic viruses like measles, rubella, varicella zoster or herpes simple shows a higher frequency in MS than any other chronic disease (MRZH reaction). We report the intrathecal, polyspecific and oligoclonal immune response in patients with definitive MS. CSF and serum were tested for Albumin, IgG, IgM and IgA by standard immunochemical nephelometry assay while virus specific antibodies in CSF and serum samples were evaluated by ELISA and calculated as antibody index. Oligoclonal IgG by, isoelectric focusing was detectable in all patients. A differential pattern of combined antibody index against neurotropic virus was observed. The largest frequency of a single species in the oligoclonal immune response was for measles antibodies, while the antibody response was found as a combination with increased rubella antibody index and VZ antibody index, respectively. Our results although preliminary for our country, enrich the criteria about that MRZH reaction provide a major neuroimmunological support to MS diagnosis.

Significance of detection of oligoclonal IgG bands in diagnosing toxoplasma gondii encephalitis in AIDS

Significance of detection of oligoclonal IgG bands in diagnosing toxoplasma gondii encephalitis in AIDS

Formulas for the quantitation of intrathecal IgG production: Their validity in the presence of blood-brain barrier damage and their utility in multiple sclerosis

There are several formulas for the quantitative determination of intrathecal IgG production: Reiber and Felgenhauer's formula (IgG(loc)), the Extended IgG index, Tourtellotte's formula (TOURT), Schuller and Sagar's formula (SCHULL), the IgG index, the Log IgG index, and Blennow and co-workers' formula (IGGPROD). To evaluate the utility of these formulas in the presence of blood-brain barrier (BBB) damage, we present the results from a study of serum and cerebrospinal fluid (CSF) samples from 125 healthy individuals, 18–88 years of age; 1072 consecutive patients without oligoclonal IgG bands (OCBs) in the CSF, 683 without BBB damage (CSF/S: albumin ratio < 9.8) and 389 with BBB damage (CSF/S albumin ratio 9.8–30); and 106 patients with definite multiple sclerosis (MS). The relation between the CSF/S albumin ratio and the CSF/S IgG ratio was remarkably linear in both healthy individuals ( r = 0.95; P < 0.0001) and patients without oligoclonal bands in the CSF ( r = 0.95; P < 0.0001). Therefore, IgG(loc) and the Extended IgG index, two formulas based on a nonlinear relation between the CSF/S albumin ratio and the CSF/S IgG ratio, yielded biased results (lower values) in the presence of BBB damage. TOURT and SCHULL also yielded biased (higher) values in the presence of BBB damage, probably because of incorrect constants in these formulas. There were no significant correlations between the CSF/S albumin ratio (i.e. the BBB function) and the IgG index or the Log IgG index, two dimensionless quotients for the detection of intrathecal IgG production, or between the CSF/S albumin ratio and IGGPROD, an empirical formula for the determination of intrathecal IgG production in mg/l. These results show that the IgG index, the Log IgG index and IGGPROD all give valid quantitative determinations of intrathecal IgG production even in the presence of BBB damage. The percentage of MS patients with a formula value higher than the mean + 2 SD of the control value was identical for the IgG index, the Log IgG index and IGGPROD (92%), and similar to the number of MS patients exhibiting OCBs in their CSF (96%). These results suggest that the quantitative determination of intrathecal IgG production is almost as sensitive as the qualitative detection of OCBs for demonstration of intrathecal IgG production.

Detection of oligoclonal IgG bands in cerebrospinal fluid by immunofixation after isoelectric focusing on polyacrylamide gels with the PhastSystem.

This paper describes the detection of oligoclonal bands of IgG immunoglobulins in cerebrospinal fluids (CSF) using isoelectric focusing (IEF) and immunofixation with the PhastSystem equipment. The proteins were separated by IEF on polyacrylamide gels, pH 3-9. The IgG immunoglobulins were then fixed by overlaying the gel with antihuman IgG antiserum. Fixed immunoglobulins were detected with silver staining in the PhastSystem Development Unit. One microliter of serum or CSF containing 15 mg/L of IgG was found to be optimal. The procedure had good reproducibility both with regard to IEF and silver staining of the fixed immunoglobulins. The manpower need was reduced, and the procedure has been found to be useful in the clinical laboratory.

Comparison of oligoclonal immunoglobulin G bands in multiple sclerosis cerebrospinal fluid by immunoblotting and immunofixation

We previously analyzed unconcentrated cerebrospinal fluid (CSF) from patients with multiple sclerosis (MS) and other neurologic diseases by isoelectric focusing in agarose gel. We have now developed an immunoblot method for detection of oligoclonal IgG bands in unconcentrated MS CSF. The oligoclonal IgG band patterns seen after immunoblotting were compared with those of conventional immunofixation. Although immunoblotting was found to be rapid the resolution and intensity of oligoclonal IgG bands were somewhat better after immunofixation. Since immunofixation is simpler than immunoblotting, we recommend that clinical laboratories use immunofixation after isoelectric focusing to detect oligoclonal IgG bands in CSF.

Immunoblotting on polyvinyldifluoride improves detection of oligoclonal IgG bands in CSF.

Journal Article Immunoblotting on polyvinyldifluoride improves detection of oligoclonal IgG bands in CSF. Get access A Nespolo, A Nespolo Search for other works by this author on: Oxford Academic Google Scholar G Bianchi, G Bianchi Search for other works by this author on: Oxford Academic Google Scholar A Salmaggi, A Salmaggi Search for other works by this author on: Oxford Academic Google Scholar D Cerrato D Cerrato Search for other works by this author on: Oxford Academic Google Scholar Clinical Chemistry, Volume 33, Issue 9, 1 September 1987, Pages 1669–1670, https://doi.org/10.1093/clinchem/33.9.1669 Published: 01 September 1987

Comparison of polyacrylamide and agarose support media for detection of oligoclonal IgG bands in CSF

Parallel samples of cerebrospinal fluid (CSF) from 120 patients clinically suspected of having multiple sclerosis were electrophoresed using agarose or 7% polyacrylamide. 57 samples showed oligoclonal IgG patterns in polyacrylamide but only 34 showed bands after concentration and electrophoresis in agarose. The concentrations of IgG and albumin were also measured in each sample and IgG/albumin ratios calculated. The sensitivities of the three methods for the detection of intrathecally synthesised IgG were compared. Electrophoresis in 7% polyacrylamide gave the highest proportion of significant results. In an attempt to increase the sensitivity of the agarose method, silver training was performed on unconcentrated samples run in agarose. This did not alter the pattern of electrophoresis in samples containing oligoclonal IgG, but the sensitivity of the technique was lower than that of polyacrylamide.

Detection of oligoclonal IgG bands in unconcentrated CSF by isoelectric focusing in agarose gel and silver staining

We analyzed 102 unconcentrated cerebrospinal fluids from a variety of neurologic diseases for oligoclonal IgG bands by isoelectric focusing in agarose gel followed by staining with silver. Ten to 12 μl of cerebrospinal fluid containing 0.4–0.8 μg of IgG was found to be optimum. Cerebrospinal fluid from 38 of 40 patients with clinically definite multiple sclerosis, 6 of 9 with suspected multiple sclerosis and 8 of 53 patients with other neurologic diseases had oligoclonal IgG bands by IEF in agarose followed by immunofixation. The commercial system employed here is a simple sensitive and rapid method for detection of oligoclonal bands in unconcentrated cerebrospinal fluid of patients with multiple sclerosis.