Isoelectric focusing followed by affinity immunoblotting to detect monoclonal free light chains in monoclonal gammopathies: Comparison with immunofixation electrophoresis and free light chain ratio.

IF 2.1 4区 医学 Q3 MEDICAL LABORATORY TECHNOLOGY
Annals of Clinical Biochemistry Pub Date : 2024-07-01 Epub Date: 2024-02-07 DOI:10.1177/00045632231221439
David Zeman, Martin Štork, Lenka Švancarová, Marek Borský, Michaela Pospíšilová, Zdeněk Adam, Miroslava Beňovská, Luděk Pour
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引用次数: 0

Abstract

Background: Isoelectric focusing (IEF) is a method with an exquisite resolution, and coupled with affinity immunoblotting (AIB), it can provide superior sensitivity to detect monoclonal free light chains (FLC).

Methods: We tested the hypothesis that IEF/AIB is more sensitive and specific for monoclonal FLC detection in serum and urine samples than conventional methods, that is, electrophoresis (ELP), immunofixation (IF) and serum FLC ratio assessment. Investigation included 107 samples of 68 patients, among which 21 multiple myeloma patients were recently tested for minimal residual disease and 18 patients with AL amyloidosis.

Results: Monoclonal FLC were detected by IEF/AIB in 37% of serum samples negative for monoclonal FLC on ELP/IF. As for urine samples, significant advantage of the IEF/AIB over ELP/IF was not demonstrated. Considering both serum and urine results, IEF/AIB definitely revealed monoclonal FLC in 20/83 (24%) of ELP/IF-negative samples. FLC ratio was abnormally high (>1.65) in all 11 patients definitely positive for monoclonal FLC kappa by IEF/AIB but also in 16/47 (34%) IEF/AIB-negative samples. Abnormally low values (<0.26) were found only in 10/28 samples (36%) positive for monoclonal FLC lambda. Appropriate use of renal FLC ratio reference range reduced the number of presumably false positives (6/47, i.e. 13%) but not false negatives (17/28, i.e. 61%).

Conclusions: The IEF/AIB method is more sensitive than IF and might be used in patients with negative IF results before deciding whether to proceed to minimal residual disease testing.

等电聚焦后进行亲和免疫印迹法检测单克隆丙种球蛋白病中的单克隆游离轻链。与免疫固定电泳和游离轻链比率进行比较。
背景:等电聚焦(IEF)是一种分辨率极高的方法,与亲和免疫印迹(AIB)相结合,可提供检测单克隆游离轻链(FLC)的更高灵敏度:方法:我们对 IEF/AIB 检测血清和尿液样本中的单克隆游离轻链的灵敏度和特异性优于电泳(ELP)、免疫固定(IF)和血清游离轻链比值评估等传统方法这一假设进行了测试。调查包括 68 名患者的 107 份样本,其中 21 名多发性骨髓瘤患者最近接受了最小残留病检测,18 名 AL 淀粉样变性患者:结果:在ELP/IF检测单克隆FLC阴性的血清样本中,有37%通过IEF/AIB检测到了单克隆FLC。至于尿液样本,IEF/AIB与ELP/IF相比没有明显优势。考虑到血清和尿液结果,在 20/83 例(24%)ELP/IF 阴性样本中,IEF/AIB 确实显示了单克隆 FLC。在所有 11 例通过 IEF/AIB 检测单克隆 FLC kappa 呈阳性的患者中,FLC 比值异常偏高(>1.65),但在 16/47 例(34%)IEF/AIB 阴性样本中,FLC 比值也异常偏高(>1.65)。异常低值(结论:IEF/AIB 方法比 IF 更敏感,可用于 IF 结果为阴性的患者,然后再决定是否进行最小残留病检测。
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来源期刊
Annals of Clinical Biochemistry
Annals of Clinical Biochemistry Biochemistry, Genetics and Molecular Biology-Clinical Biochemistry
CiteScore
5.20
自引率
4.50%
发文量
61
期刊介绍: Annals of Clinical Biochemistry is the fully peer reviewed international journal of the Association for Clinical Biochemistry and Laboratory Medicine. Annals of Clinical Biochemistry accepts papers that contribute to knowledge in all fields of laboratory medicine, especially those pertaining to the understanding, diagnosis and treatment of human disease. It publishes papers on clinical biochemistry, clinical audit, metabolic medicine, immunology, genetics, biotechnology, haematology, microbiology, computing and management where they have both biochemical and clinical relevance. Papers describing evaluation or implementation of commercial reagent kits or the performance of new analysers require substantial original information. Unless of exceptional interest and novelty, studies dealing with the redox status in various diseases are not generally considered within the journal''s scope. Studies documenting the association of single nucleotide polymorphisms (SNPs) with particular phenotypes will not normally be considered, given the greater strength of genome wide association studies (GWAS). Research undertaken in non-human animals will not be considered for publication in the Annals. Annals of Clinical Biochemistry is also the official journal of NVKC (de Nederlandse Vereniging voor Klinische Chemie) and JSCC (Japan Society of Clinical Chemistry).
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