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使用抗游离轻链抗血清通过增强敏感性改良免疫固定电泳自动检测游离单克隆轻链。

Automated detection of free monoclonal light chains by enhanced-sensitivity modified immunofixation electrophoresis with antisera against free light chains.

作者信息

Singh Gurmukh, Saldaña Emily J, Spencer Jeff, Bollag Roni J

机构信息

Department of Pathology, Medical College of Georgia at Augusta University, Augusta, GA, United States.

Helena Laboratories Inc, Beaumont, TX, United States.

出版信息

Lab Med. 2025 Sep 8;56(5):536-540. doi: 10.1093/labmed/lmaf014.

DOI:10.1093/labmed/lmaf014
PMID:40414711
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12417074/
Abstract

INTRODUCTION

About one-third of multiple myelomas produce excess free monoclonal light chains. Detection of monoclonal light chains is important for diagnosis, prognosis, and monitoring of such lesions. A previously described method for detection of monoclonal light chains in serum required multiple manual wash steps. Even though the method has sensitivity similar to that of mass spectrometry, the manual wash steps were a hindrance to the method's widespread use.

METHODS

To mitigate the laborious nature of the previous method, the SPIFE Nexus instrument (Helena Laboratories) was modified to automate the sample application, electrophoretic separation, antibody application, washing and blotting steps needed for removal of background proteins. Background noise was mitigated by modifying the wash buffer by adding a detergent. This revised automated electrophoresis protocol was tested in parallel with the previously described method.

RESULTS

The sensitivity and specificity of the modified method using antisera to free light chains from 2 sources was comparable to the parameters of the previously described method without the need for manual manipulation.

DISCUSSION

The automated protocol employing the SPIFE Nexus instrument and incorporating antisera to free light chains is suitable for routine use in clinical laboratories in an automated, enhanced-sensitivity assay for monoclonal light chains with no need for manual manipulation.

摘要

引言

约三分之一的多发性骨髓瘤会产生过量游离单克隆轻链。检测单克隆轻链对于此类病变的诊断、预后评估及监测至关重要。先前描述的一种血清中单克隆轻链检测方法需要多个手动洗涤步骤。尽管该方法的灵敏度与质谱法相似,但手动洗涤步骤阻碍了其广泛应用。

方法

为减轻先前方法的繁琐程度,对SPIFE Nexus仪器(海伦娜实验室)进行了改进,以实现样品加样、电泳分离、抗体加样、去除背景蛋白所需的洗涤和印迹步骤的自动化。通过添加去污剂对洗涤缓冲液进行改良,降低背景噪音。将修订后的自动电泳方案与先前描述的方法进行平行测试。

结果

使用来自2种来源的游离轻链抗血清的改良方法的灵敏度和特异性与先前描述方法的参数相当,无需手动操作。

讨论

采用SPIFE Nexus仪器并结合游离轻链抗血清的自动方案适用于临床实验室的常规使用,可在无需手动操作的情况下,以自动化、高灵敏度的检测方法检测单克隆轻链。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9b0b/12417074/a5ef2893e0aa/lmaf014_fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9b0b/12417074/fb6cf7df7cad/lmaf014_fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9b0b/12417074/5eab26dacfff/lmaf014_fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9b0b/12417074/a5ef2893e0aa/lmaf014_fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9b0b/12417074/fb6cf7df7cad/lmaf014_fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9b0b/12417074/5eab26dacfff/lmaf014_fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9b0b/12417074/a5ef2893e0aa/lmaf014_fig3.jpg

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