scholarly journals Our Laboratory Experience: Comparison of Capillary Electrophoresis/Immunosubtraction and Agarose Gel/Immunofixation

2020 ◽  
Vol 2 (7) ◽  
pp. 267-277
Author(s):  
Massimo Pieri ◽  
Flaminia Tomassetti ◽  
Caterina Iodice ◽  
Rosa Piazzolla ◽  
Elena Riboldi ◽  
...  
Keyword(s):  
Serum Protein ◽  
Wide Spectrum ◽  
Protein Electrophoresis ◽  
Serum Protein Electrophoresis ◽  
Common Denominator ◽  
Agarose Gel ◽  
Serum Samples ◽  
Suitable Alternative ◽  
Immunofixation Electrophoresis ◽  
Monoclonal Proteins

Abstract. Monoclonal gammopathies represent a wide spectrum of related diseases. The common denominator is the presence of a monoclonal protein in the serum or urine, which can be in the form of intact immunoglobulin, immunoglobulin fragments, and/or free light chains (κ and λ). We can detect these abnormalities by immunofixation electrophoresis (IFE) in which specific antibodies are overlaid after electrophoresis and the corresponding immunoglobulin. In our study, we compared gel-based and capillary-based serum protein electrophoresis methods to identify and characterize monoclonal immunoglobulins in serum samples. We analyzed 500 serum samples by Sebia Hydragel agarose gel electrophoresis (AGE)/immunofixation electrophoresis (IFE) and CAPILLARYS 2 capillary zone electrophoresis (CZE)/immunosubtraction (IS). AGE/IFE and CZE/IS had an overall agreement of 98% on serum protein electrophoresis. In our hands, AGE/IFE and CZE/IS had the same specificity for detection of monoclonal proteins; however, CZE/IS seems to be more sensitive than AGE/IFE for the detection of some critical cases. However, CZE/IS is an analytically suitable alternative to AGE/IFE, although laboratories should need to combine, for singular samples, both electrophoretic methods in their clinical routine. A combined use of AGE/IFE and CZE/IS is necessary to lead to an accurate diagnosis and clinically error-free results.

The Detection by Immunofixation Electrophoresis (IFE) of Monoclonal Proteins in Veteran Patients with Hypogammaglobulinemia on Negative Serum Protein Electrophoresis, the Veteran Affairs Medical Center (VAMC) Experience

2020 ◽  
Vol 154 (Supplement_1) ◽  
pp. S89-S89
Author(s):  
J M Petersen ◽  
M Litman ◽  
R Millili ◽  
D Jhala
Keyword(s):  
Serum Protein ◽  
Medical Center ◽  
Protein Electrophoresis ◽  
Serum Protein Electrophoresis ◽  
Time Analysis ◽  
Improvement Project ◽  
Immunofixation Electrophoresis ◽  
M Spike ◽  
Monoclonal Proteins ◽  
First Time

Abstract Introduction/Objective Serum protein electrophoresis (SPEP) is the backbone laboratory test for the detection of abnormal monoclonal proteins. However, IFE is a sensitive assay that can sometimes detect monoclonal proteins even when the corresponding SPEP is negative. The fact that IFE is more sensitive than SPEP combined with the need to avoid overutilization of IFE has led to published algorithms for guidance. Hypogammaglobulinemia in a new patient has been recognized in these algorithms as a reason to reflex to IFE when SPEP is negative, though studies on veteran patients are sparse. Therefore, this QA study of the percentage of positive IFEs in negative new SPEP veteran patients with hypogammaglobulinemia was undertaken to ensure reflex IFEs would still be indicated. Methods As part of a quality assurance/quality improvement project, a retrospective Vista/Fileman search of all SPEPs with IFE performed from January 2017 to February 2019 was undertaken to identify cases of SPEPs showing hypogammaglobulinemia (<0.7 g/dL). Diagnostic comments were then analyzed to identify cases of hypogammaglobulinemia along with the M-spike (<0.1 g/dL) to identify negative SPEPs. Only those cases that were consistent with first time hypogammaglobulinemia without an obvious M-spike were included and tabulated for calculations. Any result that was not negative for a monoclonal band was considered as positive. Results There were a total of 194 specimens that met the criteria of SPEP with hypogammaglobulinemia and standard comments consistent with first time analysis and without an obvious M-spike on SPEP. Out of these 194 specimens, 45 had a positive result, either as a monoclonal band comigrating with the beta protein peak or as a very faint gamma or beta monoclonal band. This represented approximately 23% of the specimens, about double the literature published rate for the non-veteran population. Conclusion The performance of IFE on new hypogammaglobulinemia veteran patients appears to be indicated like previously published algorithms and is supported by the fact that about double (23%), compared to the non-veteran population, had positive IFEs despite negative SPEPs. IFE is a helpful tool for new hypogammaglobulinemia patients for the detection of monoclonal proteins despite negative SPEPs.

When Patients Have Neuropathy: Frequency Of A Positive Immunofixation Electrophoresis (IFE) For Monoclonal Protein When The Serum Protein Electrophoresis (SPEP) Is Negative, The Veteran Affairs Medical Center (VAMC) Experience

2020 ◽  
Vol 154 (Supplement_1) ◽  
pp. S88-S88
Author(s):  
J M Petersen ◽  
M Litman ◽  
R Millili ◽  
D Jhala
Keyword(s):  
Serum Protein ◽  
Limit Of Detection ◽  
Medical Center ◽  
Protein Electrophoresis ◽  
Serum Protein Electrophoresis ◽  
Record System ◽  
Improvement Project ◽  
Veteran Affairs ◽  
Immunofixation Electrophoresis ◽  
Monoclonal Proteins

Abstract Introduction/Objective Although serum protein electrophoresis (SPEP) is the backbone of testing to determine the presence of monoclonal proteins, immunofixation electrophoresis (IFE) is more sensitive with the ability to pick up fainter bands or bands comigrating with the normal beta protein peak. Given the costs of performing additional IFE and the known increased sensitivity, algorithms for reflex IFE published in the literature have stated that unexplained neuropathy would be an indication for reflex IFE even if the SPEP appears negative in a new patient. However, literature on exactly how many of these reflex IFE’s end up positive is sparse. Therefore, we present the Veteran Affairs Medical Center (VAMC) experience in a veteran population. Methods As part of a quality assurance/improvement project, retrospective Vista/Fileman search of all SPEPs with IFE performed from January 2017 to February 2019 was undertaken to identify all SPEPs/IFE with the standard comment language typically used in patients with neuropathy, namely a preliminary diagnosis of no monoclonal bands on SPEP with an IFE to confirm this diagnosis. All SPEPs and IFEs had undergone review by a pathologist prior to signout which includes a chart review in the computerized patient record system (CPRS) to rule out the documented presence of unexplained neuropathy in new patients who have not previously had IFE workup. Results There were 129 SPEP/IFE cases identified. Of these 129, the overwhelming majority or 123 specimens (95%) were negative on IFE. However, there were 6 specimens (5%) that demonstrated a band on IFE without a band being identified on the SPEP. These bands were in the beta region or very faintly located in the gamma region at or below the limit of detection of the SPEP Conclusion The performance of IFE for patients with unexplained neuropathy would, in the veteran population at the VAMC, identify a monoclonal band in about 1/20 cases. This provides support for the published algorithms advocating for performing IFE on negative SPEP specimens from patients with unexplained neuropathy, as this finding might be important and would otherwise have been missed if just the SPEP was performed alone.

Measurement of Monoclonal Immunoglobulin Protein Concentration in Serum Protein Electrophoresis: Comparison of Automated vs Manual/Human Readings

2019 ◽  
Vol 51 (3) ◽  
pp. 252-258 ◽  
Author(s):  
Alex Clavijo ◽  
Nathan Ryan ◽  
Hongyan Xu ◽  
Gurmukh Singh
Keyword(s):  
Serum Protein ◽  
Protein Concentration ◽  
Protein Electrophoresis ◽  
Serum Protein Electrophoresis ◽  
Response To Treatment ◽  
Computer Assisted ◽  
Positive Bias ◽  
Monoclonal Immunoglobulin ◽  
Monoclonal Immunoglobulins ◽  
Immunofixation Electrophoresis

Abstract Background Protein concentration of monoclonal immunoglobulin in plasma-cell myeloma/multiple myeloma provides an estimate of the tumor mass and allows for monitoring of the response to treatment. Accurate and reproducible estimates of the monoclonal immunoglobulin concentration are important for patient care. Objective To address the optimum method for estimation of the concentration of monoclonal immunoglobulins. Methods Serum protein electrophoresis and immunofixation electrophoresis were conducted by using the Helena SPIFE Touch instrument. Estimation of the protein concentration of monoclonal immunoglobulin in the gamma region by computer-assisted reading was compared with the reading by technologists and pathology residents, in 300 gels. The data were compared using t-testing and analysis of variance. Results Computer-generated readings had a consistent positive bias. The correlation coefficient of the average reading by technologists and residents with the computer generated value was 0.997. The average positive bias by the computer reading was 0.29 g per dL. The intercept on the regression analysis was 0.22 g per dL. The reading by the computer was significantly higher than each of the human-interpreted readings. The readings by the 3 human groups were not significantly different amongst them. The main reason for the higher reading by the computer was inclusion of a greater area on the anodal size of the peak on the densitometric scan. Conclusions Human- and computer-interpreted readings of the protein concentration of monoclonal immunoglobulin have a high degree of correlation. The consistent positive bias by the computer reading occurred due to inclusion of a greater area of the densitometric scan on the anodal side of the peak. We suggest that vendors should adjust such computer programs to provide readings comparable to those generated by expert humans. We recommend manual delineation of the monoclonal peaks for measuring the concentration of monoclonal immunoglobulins.

scholarly journals Agarose gel serum protein electrophoresis in cats with and without lymphoma and preliminary results of tandem mass fingerprinting analysis

2011 ◽  
Vol 40 (2) ◽  
pp. 159-173 ◽  
Author(s):  
Magda Gerou-Ferriani ◽  
Alix R. McBrearty ◽  
Richard J. Burchmore ◽  
Kamburapola G.I. Jayawardena ◽  
P. David Eckersall ◽  
...  
Keyword(s):  
Serum Protein ◽  
Protein Electrophoresis ◽  
Serum Protein Electrophoresis ◽  
Tandem Mass ◽  
Agarose Gel ◽  
Preliminary Results ◽  
Fingerprinting Analysis

scholarly journals Simple thrombin-based method for eliminating fibrinogen interference in serum protein electrophoresis of haemodialysed patients

2020 ◽  
Vol 30 (2) ◽  
pp. 265-271
Author(s):  
Anamarija Rade ◽  
Anamarija Đuras ◽  
Irena Kocijan ◽  
Patricija Banković Radovanović ◽  
Ana Turčić
Keyword(s):  
Serum Protein ◽  
Venous Catheter ◽  
Protein Electrophoresis ◽  
Serum Protein Electrophoresis ◽  
Blood Collection ◽  
Serum Samples ◽  
Becton Dickinson ◽  
Vascular Access Devices ◽  
Hospital Protocols ◽  
Clot Activator

Introduction: Serum samples of haemodialysed patients collected through vascular access devices, e.g. central venous catheter (CVC) can contain residual heparin, which can cause incomplete clotting and consequently fibrinogen interference in serum protein electrophoresis (SPE). We hypothesized that this problem may be overcome by addition of thrombin and aimed to find a simple thrombin-based method for fibrinogen interference removal. Materials and methods: Blood samples of 51 haemodialysed patients with CVC were drawn through catheter into Clot Activator Tube (CAT) and Rapid Serum Tube Thrombin (RST) vacutainers (Becton Dickinson, New Jersey, USA) following the routine hospital protocols and analysed with gel-electrophoresis (Sebia, Lisses, France). Samples were redrawn in the CAT tubes and re-analysed after being treated with thrombin using two methods: transferring CAT serum into RST vacutainer and treatment of CAT serum with fibrinogen reagent (Multifibren U, Siemens, Marburg, Germany). Results: Direct blood collection in RST proved to be slightly more efficient than CAT in removing the interfering band in beta fraction (CAT removed 6/51 and RST removed 12/51, P = 0.031). Transferring CAT serum into the RST vacutainer proved to be more efficient for subsequent removal of interfering band from CAT serum than the addition of fibrinogen reagent (39/45 vs. 0/45 samples with efficiently removed interfering band, P < 0.001). Conclusion: Fibrinogen interference caused by incomplete clotting because of residual heparin can be overcome by addition of thrombin. Transferring CAT serum into the RST vacutainer was the most efficient method.

Test Utilization of Serum Protein Electrophoresis and Immunofixation Electrophoresis

2019 ◽  
Vol 152 (Supplement_1) ◽  
pp. S146-S147
Author(s):  
Roula Katerji ◽  
Tamera Paczos ◽  
Li Liu
Keyword(s):  
Multiple Myeloma ◽  
Serum Protein ◽  
Active Treatment ◽  
Cost Effective ◽  
Protein Electrophoresis ◽  
Serum Protein Electrophoresis ◽  
Efficient Test ◽  
Testing Frequency ◽  
Immunofixation Electrophoresis ◽  
Changes Over Time

Abstract Objectives Serum protein electrophoresis (SPE) and immunofixation electrophoresis (IFE) are commonly used to screen and monitor monoclonal gammopathies. Currently, there are no consensus guidelines on optimal testing frequency leading to overutilization. Here we examined the testing frequencies of SPE and IFE in our institution to provide an evidence-based perspective on efficient test utilization. Methods We retrospectively reviewed all SPE and IFE tests performed in 2018. Ordering patterns and testing frequencies were analyzed. In cases with more than monthly repeats, electronic medical records were reviewed to follow the result changes over time. Results There were 10,054 SPE and 4,248 IFE orders in 2018. The 4,248 IFE cases represented 2,439 patients, among whom 104 patients (4.3%) had IFE repeated more frequently than every 2 months and 50 patients (2%) more frequently than monthly. The 10,054 SPE cases represented 5,472 patients; 127 patients (2.3%) had SPE performed more than 12 times. Rare cases (0.1%) had SPE and IFE repeated every 1 to 2 weeks. Most IFE tests were ordered together with SPE (89% of all IFE orders), among which 28% of cases had normal SPE findings. Among the cases with more than monthly SPE and IFE tests, IFE results showed meaningful interpretation changes in a minimum period of 2 to 3 months; 35% cases had no IFE interpretation changes throughout the year. In contrast, during active treatment period of multiple myeloma, SPE detected paraprotein level change weekly. Conclusion IFE is overutilized and monthly monitoring does not add value even during active treatment of multiple myeloma. Our results support the need for the development of testing frequency guidelines to avoid overutilization and provide more cost-effective patient care.

A comparison between high resolution serum protein electrophoresis and screening immunofixation for the detection of monoclonal gammopathies in serum

2018 ◽  
Vol 56 (2) ◽  
pp. 256-263 ◽  
Author(s):  
Joel Smith ◽  
Geoffrey Raines ◽  
Hans-Gerhard Schneider
Keyword(s):  
High Resolution ◽  
Sensitivity And Specificity ◽  
Serum Protein ◽  
Light Chain ◽  
Monoclonal Gammopathy ◽  
Protein Electrophoresis ◽  
Serum Protein Electrophoresis ◽  
Agarose Gel ◽  
Serum Free ◽  
Monoclonal Gammopathies

Abstract Background: There are a variety of initial laboratory tests or combinations of tests that can be performed when a monoclonal gammopathy is suspected including serum protein electrophoresis (SPEP), urine protein electrophoresis (UPEP), serum immunofixation (IFE) and serum free light chain assays. Some groups have recently used simplified “screening” IFE methods for the detection of monoclonal gammopathies leveraging the greater sensitivity of IFE over SPEP alone to improve the detection of monoclonal gammopathies. These screening techniques have been predominantly evaluated against lower resolution agarose gel electrophoresis techniques. Methods: In this study we evaluated the diagnostic performance of the combined κ and λ light chain screening immunofixation (CLIF) in comparison to serum protein electrophoresis on a high-resolution (Sebia Hydragel 15 HR) agarose gel system. Each gel was interpreted by three adjudicators. A total of 156 patient samples were analysed. Adjudicated diagnoses based on the screening techniques were compared against the results of high resolution serum protein electrophoresis and high resolution standard immunofixation performed during routine laboratory operation. Where standard immunofixation was not performed a combination of a review of medical records, serum free light chains, UPEP and bone marrow aspirate and trephine and subsequent standard immunofixation and protein electrophoresis results where available were used to confirm the absence of a monoclonal gammopathy. Results: In this cohort a total of 65 (41%) patients had a paraprotein confirmed by standard immunofixation. HR SPEP had a sensitivity and specificity of 95% and 85%, respectively, while CLIF had a sensitivity and specificity of 88% and 97%, respectively. Conclusions: Overall we found that high-resolution gel serum protein electrophoresis using a Sebia Hydragel 15 HR system was more sensitive than a screening immunofixation method (CLIF) for the detection of paraproteins in patient serum in this patient cohort. The drawback of the greater sensitivity of HR SPEP was a higher false positive rate requiring an increased utilisation of follow up immunofixation electrophoresis.

Laboratory testing requirements for diagnosis and follow-up of multiple myeloma and related plasma cell dyscrasias

2016 ◽  
Vol 54 (6) ◽  
Author(s):  
Maria A.V. Willrich ◽  
Jerry A. Katzmann
Keyword(s):  
Multiple Myeloma ◽  
Serum Protein ◽  
Plasma Cell ◽  
Light Chain ◽  
Cell Clone ◽  
Protein Electrophoresis ◽  
Free Light Chain ◽  
Molecular Weights ◽  
Immunofixation Electrophoresis ◽  
Monoclonal Proteins

AbstractMonoclonal immunoglobulins are markers of plasma cell proliferative diseases and have been described as the first (and perhaps best) serological tumor marker. The unique structure of each monoclonal protein makes them highly specific for each plasma cell clone. The difficulties of using monoclonal proteins for diagnosing and monitoring multiple myeloma, however, stem from the diverse disease presentations and broad range of serum protein concentrations and molecular weights. Because of these challenges, no single test can confidently diagnose or monitor all patients. Panels of tests have been recommended for sensitivity and efficiency. In this review we discuss the various disease presentations and the use of various tests such as protein electrophoresis and immunofixation electrophoresis as well as immunoglobulin quantitation, free light chain quantitation, and heavy-light chain quantitation by immuno-nephelometry. The choice of tests for inclusion in diagnostic and monitoring panels may need to be tailored to each patient, and examples are provided. The panel currently recommended for diagnostic screening is serum protein electrophoresis, immunofixation electrophoresis, and free light chain quantitation.

Serum Free Kappa to Free Lambda Ratios as an Adjunct to Serum Protein Electrophoresis for the Detection of Monoclonal Proteins in the Serum.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4856-4856
Author(s):  
Arthur R. Bradwell ◽  
Jean Garbincius ◽  
Earle W. Holmes
Keyword(s):  
Serum Protein ◽  
Light Chain ◽  
Protein Electrophoresis ◽  
Free Light Chain ◽  
Serum Protein Electrophoresis ◽  
Serum Free ◽  
Monoclonal Protein ◽  
Serum Free Light Chain ◽  
Monoclonal Proteins

Abstract Serum free light chain measurements have been shown to be useful in the diagnosis and monitoring of patients with monoclonal gammopathies. The present study was undertaken to evaluate the effect of adding the measurement of serum free light chain kappa to lambda ratios to the serum protein electrophoresis evaluation that we typically use as an initial screen for the detection of monoclonal proteins. We retrospectively tested 347 consecutive samples from individuals who had no previous history of plasma cell dyscrasia and had not previously had a serum or urine electrophoresis or immunofixation electrophoresis test at our institution. The quantitative serum protein electrophoresis test that was ordered was performed using Hydragel Beta 1- Beta 2 gels and Hydrasis instrument (Sebia, Inc., Norcross, GA). The protein content of the electrophoresis zones were quantitated by scanning densitometry and the electrophoresis pattern of each sample was qualitatively examined for abnormal bands and suspicious findings by a single, experienced observer. Serum free light chain concentrations and the serum free light chain kappa to lambda ratios were determined using the Freelite Human Kappa and Lambda Kits (The Binding Site Ltd, Birmingham, UK) and the Immage analyzer (Beckman Coulter Inc., Brea, CA). The serum free light chain kappa to lambda ratios were outside the reference interval (0.25 to1.65) in 23 of the samples. Ten abnormal ratios were observed among a group of 57 samples that had either positive or suspicious qualitative evaluations for the presence of a restriction or that demonstrated hypo-gammaglobulinemia. Both abnormalities led to recommendations for follow-up testing, which confirmed the presence of a monoclonal protein in 21 of the samples. Six abnormal ratios were observed among a group of 159 specimens that had quantitative abnormalities in albumin or one or more of globulin fractions (hypo-gammaglobulinemia excepted) and normal qualitative evaluations. Seven abnormal ratios were observed among a group of 131 samples that had normal quantitative results and normal qualitative evaluations. Follow-up testing is not usually recommended for serum protein electrophoresis results like those in the latter two groups. We found that the addition of the serum free light chain kappa to lambda ratio to the serum protein electrophoresis test increased the number of abnormal screens that would have required further clinical and/or laboratory evaluation by 23%(i.e. from 57 to 70). Given the high specificity of the serum free light chain kappa to lambda ratio for monoclonal light chains, the additional 13 abnormal samples identified by this test are expected to have a high likelihood of harboring a monoclonal protein that would have otherwise eluded detection. Pending a definitive prospective study, we estimate that the addition of a serum free light chain kappa to lambda ratio to the serum protein electrophoresis screen would increase the rate of detection of serum monoclonal proteins by as much as 1.6-fold.

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