Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou, China.
J Clin Lab Anal. 2013 Jul;27(4):277-83. doi: 10.1002/jcla.21597.
Cytokeratin 19 fragment antigen (CYFRA 21-1) is used to diagnose and monitor neoplasms. However, the main disadvantages of the currently available CYFRA 21-1 assays include heterogenous technology, being time-consuming, and having low through-put with low insensitivity. This study investigated the use of amplified luminescent proximity homogeneous immunoassay (AlphaLISA) for the quantization of CYFRA 21-1 in human serum.
The AlphaLISA kit was developed based on AlphaScreen detection technology with two different anti-CYFRA 21-1 monoclonal antibodies. One was coated on AlphaLISA acceptor beads and the other was biotinylated. Donor beads were coated with streptavidin. The test conditions were optimized and analytical performance was studied.
The measurement range of AlphaLISA CYFRA 21-1 kit was 0.08-500 ng/ml. Assay detection limit was 0.08 ng/ml. The intra- and interassay coefficients of variation were 3.00-9.00% and 4.00-10.00%, respectively. There was no cross-reaction to alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), cancer antigen 19-9 (CA19-9), cytokeratins 8 (CK8), and cytokeratins 18 (CK18). The correlation coefficient of blood samples involved was 0.974 between CYFRA 21-1-AlphaLISA assay and a commercial electrochemiluminescence immunoassay (ECLIA) CYFRA 21-1 kit (Roche).
The AlphaLISA CYFRA 21-1 kit developed in this study had favorable performance characteristics for clinical application with acceptable analytical sensitivity, specificity, and accuracy.
细胞角蛋白 19 片段抗原（CYFRA 21-1）用于诊断和监测肿瘤。然而，目前可用的 CYFRA 21-1 检测方法的主要缺点包括技术异质性、耗时、高通量低且灵敏度低。本研究探讨了扩增发光邻近均相免疫测定（AlphaLISA）在人血清中定量检测 CYFRA 21-1 的应用。
基于 AlphaScreen 检测技术，使用两种不同的抗 CYFRA 21-1 单克隆抗体开发了 AlphaLISA 试剂盒。一种包被在 AlphaLISA 接受珠上，另一种生物素化。供体珠包被有链霉亲和素。优化了测试条件并研究了分析性能。
AlphaLISA CYFRA 21-1 试剂盒的测量范围为 0.08-500ng/ml。检测限为 0.08ng/ml。批内和批间变异系数分别为 3.00-9.00%和 4.00-10.00%。与甲胎蛋白（AFP）、癌胚抗原（CEA）、神经元特异性烯醇化酶（NSE）、癌抗原 19-9（CA19-9）、细胞角蛋白 8（CK8）和细胞角蛋白 18（CK18）无交叉反应。涉及的血液样本的相关系数在 0.974 之间，AlphaLISA 法和商业电化学发光免疫分析法（ECLIA）CYFRA 21-1 试剂盒（罗氏）之间（Roche）。
本研究开发的 AlphaLISA CYFRA 21-1 试剂盒具有良好的临床应用性能，具有可接受的分析灵敏度、特异性和准确性。
