Hamaguchi Yasuhito, Kuwana Masataka, Takehara Kazuhiko
Department of Dermatology, Faculty of Medicine, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa, 920-8641, Japan.
Department of Allergy and Rheumatology, Nippon Medical School Graduate School of Medicine, Tokyo, Japan.
Clin Rheumatol. 2020 Nov;39(11):3489-3497. doi: 10.1007/s10067-020-04973-0. Epub 2020 May 8.
INTRODUCTION/OBJECTIVES: A line blot (LB) assay is a multi-analyte platform capable of simultaneously detecting multiple anti-nuclear antibody specificities. Here, we evaluated the performance of a commercial LB assay developed for the identification of myositis- or systemic sclerosis (SSc)-related autoantibodies (autoAbs).
We screened 300 serum samples from patients with various connective tissue diseases using an LB assay and compared the results of myositis- or SSc-related autoAbs with those identified by RNA and protein immunoprecipitation (IP) assays or indirect immunofluorescence (IIF).
The IP assays revealed anti-Jo-1 Abs in 14 patients, anti-EJ Abs in 12, anti-PL-7 Abs in 8, anti-PL-12 Abs in 4, anti-Mi-2 Abs in 6, anti-SRP Abs in 8, anti-topoisomerase I Abs in 54, anti-RNA polymerase III Abs in 24, anti-U3 RNP Abs in 9, anti-Th/To Abs in 9, anti-Ku Abs in 14 and anti-hUBF Abs in 4, whereas IIF identified anti-centromere in 35. Good agreement between the IP assays and the LB assay was found only for anti-Jo-1 and anti-centromere antibodies. When a cut-off was adjusted to reconcile with the results of IP assays, the detection performance of LB assay was improved for anti-EJ, anti-PL-7, anti-PL-12, anti-SRP, anti-topoisomerase I and anti-RNA polymerase III Abs. However, the results of anti-Mi-2, anti-U3 RNP, anti-Th/To, anti-hUBF and anti-Ku Abs remained discordant between the LB assay and IP assays at all cut-off levels.
Detection of myositis- or SSc-related autoAbs using a commercial LB assay requires great caution since it can yield analytically false-positive or false-negative results. Key Points • A line blot (LB) assay is a multi-analyte platform capable of simultaneously detecting multiple antibodies with anti-nuclear specificities. • Detection of myositis- or systemic sclerosis-related autoantibodies using a commercial LB assay requires great caution since it can yield analytically false-positive or false-negative results.
引言/目的：线印迹（LB）检测是一种能够同时检测多种抗核抗体特异性的多分析物平台。在此，我们评估了一种用于鉴定肌炎或系统性硬化症（SSc）相关自身抗体（自身抗体）的商业LB检测的性能。
我们使用LB检测对300例各种结缔组织疾病患者的血清样本进行了筛查，并将肌炎或SSc相关自身抗体的检测结果与通过RNA和蛋白质免疫沉淀（IP）检测或间接免疫荧光（IIF）鉴定的结果进行了比较。
IP检测在14例患者中发现抗Jo-1抗体，12例中发现抗EJ抗体，8例中发现抗PL-7抗体，4例中发现抗PL-12抗体，6例中发现抗Mi-2抗体，8例中发现抗SRP抗体，54例中发现抗拓扑异构酶I抗体，24例中发现抗RNA聚合酶III抗体，9例中发现抗U3 RNP抗体，9例中发现抗Th/To抗体，14例中发现抗Ku抗体，4例中发现抗hUBF抗体，而IIF在35例中鉴定出抗着丝点抗体。仅在抗Jo-1和抗着丝点抗体方面，IP检测与LB检测结果具有良好的一致性。当调整临界值以与IP检测结果一致时，LB检测对抗EJ、抗PL-7、抗PL-12、抗SRP、抗拓扑异构酶I和抗RNA聚合酶III抗体的检测性能有所提高。然而，在所有临界值水平下，LB检测与IP检测在抗Mi-2、抗U3 RNP、抗Th/To、抗hUBF和抗Ku抗体的结果上仍不一致。
使用商业LB检测来检测肌炎或SSc相关自身抗体需要格外谨慎，因为它可能会产生分析性假阳性或假阴性结果。要点 • 线印迹（LB）检测是一种能够同时检测多种具有抗核特异性抗体的多分析物平台。 • 使用商业LB检测来检测肌炎或系统性硬化症相关自身抗体需要格外谨慎，因为它可能会产生分析性假阳性或假阴性结果。
