BACKGROUND: 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. METHODS: 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. RESULTS: 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). CONCLUSIONS: The AlphaLISA CYFRA 21-1 kit developed in this study had favorable performance characteristics for clinical application with acceptable analytical sensitivity, specificity, and accuracy.
BACKGROUND: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. METHODS: 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. RESULTS: 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). CONCLUSIONS: The AlphaLISA CYFRA 21-1 kit developed in this study had favorable performance characteristics for clinical application with acceptable analytical sensitivity, specificity, and accuracy.
Authors: E F Ullman; H Kirakossian; A C Switchenko; J Ishkanian; M Ericson; C A Wartchow; M Pirio; J Pease; B R Irvin; S Singh; R Singh; R Patel; A Dafforn; D Davalian; C Skold; N Kurn; D B Wagner Journal: Clin Chem Date: 1996-09 Impact factor: 8.327
Authors: Stefan Holdenrieder; Joachim Von Pawel; Thomas Duell; Knut Feldmann; Hannelore Raith; Annette Schollen; Dorothea Nagel; Petra Stieber Journal: Anticancer Res Date: 2010-05 Impact factor: 2.480
Authors: Khaled Alkotyfan; Susanne Wiegand; Hans-Helge Müller; Jochen P Windfuhr; Jochen A Werner; Andreas M Sesterhenn Journal: Anticancer Res Date: 2010-06 Impact factor: 2.480
Authors: Gero Kramer; Hamdiye Erdal; Helena J M M Mertens; Marius Nap; Julian Mauermann; Georg Steiner; Michael Marberger; Kenneth Bivén; Maria C Shoshan; Stig Linder Journal: Cancer Res Date: 2004-03-01 Impact factor: 12.701
Authors: Gabriel Lassabe; Karl Kramer; Bruce D Hammock; Gualberto González-Sapienza; Andrés González-Techera Journal: Anal Chem Date: 2018-05-01 Impact factor: 6.986
Authors: Helena Pulido-Olmo; Elena Rodríguez-Sánchez; José Alberto Navarro-García; María G Barderas; Gloria Álvarez-Llamas; Julián Segura; Marisol Fernández-Alfonso; Luis M Ruilope; Gema Ruiz-Hurtado Journal: Front Immunol Date: 2017-07-24 Impact factor: 7.561