Maria Infantino1, O Shovman2,3, B Gilburd2, M Manfredi4, V Grossi4, Maurizio Benucci5, A Damiani5, D Chimenti6, K Malyavantham7, Y Shoenfeld2,3,8. 1. Immunology and Allergology Laboratory Unit, S. Giovanni di Dio Hospital, Azienda Usl Toscana Centro, Via Torregalli, 3, 50143, Florence, Italy. maria2.infantino@uslcentro.toscana.it. 2. Zabludowitz Center for Autoimmune Diseases, Sheba Medical Center, Ramat Gan, Israel. 3. Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel. 4. Immunology and Allergology Laboratory Unit, S. Giovanni di Dio Hospital, Azienda Usl Toscana Centro, Via Torregalli, 3, 50143, Florence, Italy. 5. Rheumatology Unit, S. Giovanni di Dio Hospital, Azienda Usl Toscana Centro, Florence, Italy. 6. A.Menarini Diagnostics, Florence, Italy. 7. Trinity Biotech, Buffalo, NY, USA. 8. Incumbent of the Laura Schwarz-Kipp Chair for Research of Autoimmune Diseases, Tel Aviv University, Tel Aviv, Israel.
Abstract
INTRODUCTION/ OBJECTIVES: Accurate interpretation of DFS70 (dense fine speckled 70) and mixed antinuclear antibodies (ANAs) patterns can be challenging using conventional HEp-2 immunofluorescence (IIF) method. We evaluated a novel HEp-2 IIF substrate (HEp-2 ELITE/DFS70-KO) composed of a mixture of engineered HEp-2 devoid of the DFS70 autoantigen and conventional HEp-2 cells. The study assessed the utility of the new substrate in ANA screening and its advantages. METHOD: One thousand and five consecutive routine samples sent for ANA screening were tested on both standard HEp-2 and the HEp-2 ELITE DFS70 KO substrates (ImmuGlo ANA HEp-2 and HEp-2 ELITE/DFS70-KO, Trinity Biotech, Buffalo, NY). Anti-DFS70 antibody specificity was additionally determined by immunoblot (IB). Clinical and serological data were included in the analysis of the overall impact of the novel HEp-2 substrate on DFS pattern interpretation. RESULTS: Of the 22 cases suspected as positive for DFS pattern alone or in combination with homogeneous or speckled patterns on conventional HEp-2 cells, 17 were interpreted with a higher accuracy using the new HEp-2 ELITE method as positive for DFS70 (monospecific DFS70 (10), mixed DFS70 (7)), speckled (3), and DFS (2) patterns. CONCLUSIONS: The new substrate was not only useful in deciphering unclear mixed ANA patterns but also highly sensitive in detecting DFS70 pattern in comparison to the DFS70 positivity obtained using IB.
INTRODUCTION/ OBJECTIVES: Accurate interpretation of DFS70 (dense fine speckled 70) and mixed antinuclear antibodies (ANAs) patterns can be challenging using conventional HEp-2 immunofluorescence (IIF) method. We evaluated a novel HEp-2 IIF substrate (HEp-2 ELITE/DFS70-KO) composed of a mixture of engineered HEp-2 devoid of the DFS70 autoantigen and conventional HEp-2 cells. The study assessed the utility of the new substrate in ANA screening and its advantages. METHOD: One thousand and five consecutive routine samples sent for ANA screening were tested on both standard HEp-2 and the HEp-2 ELITE DFS70 KO substrates (ImmuGlo ANA HEp-2 and HEp-2 ELITE/DFS70-KO, Trinity Biotech, Buffalo, NY). Anti-DFS70 antibody specificity was additionally determined by immunoblot (IB). Clinical and serological data were included in the analysis of the overall impact of the novel HEp-2 substrate on DFS pattern interpretation. RESULTS: Of the 22 cases suspected as positive for DFS pattern alone or in combination with homogeneous or speckled patterns on conventional HEp-2 cells, 17 were interpreted with a higher accuracy using the new HEp-2 ELITE method as positive for DFS70 (monospecific DFS70 (10), mixed DFS70 (7)), speckled (3), and DFS (2) patterns. CONCLUSIONS: The new substrate was not only useful in deciphering unclear mixed ANA patterns but also highly sensitive in detecting DFS70 pattern in comparison to the DFS70 positivity obtained using IB.
Authors: Nancy Agmon-Levin; Jan Damoiseaux; Cees Kallenberg; Ulrich Sack; Torsten Witte; Manfred Herold; Xavier Bossuyt; Lucille Musset; Ricard Cervera; Aresio Plaza-Lopez; Carlos Dias; Maria José Sousa; Antonella Radice; Catharina Eriksson; Olof Hultgren; Markku Viander; Munther Khamashta; Stephan Regenass; Luis Eduardo Coelho Andrade; Allan Wiik; Angela Tincani; Johan Rönnelid; Donald B Bloch; Marvin J Fritzler; Edward K L Chan; I Garcia-De La Torre; Konstantin N Konstantinov; Robert Lahita; Merlin Wilson; Olli Vainio; Nicole Fabien; Renato Alberto Sinico; Pierluigi Meroni; Yehuda Shoenfeld Journal: Ann Rheum Dis Date: 2013-10-14 Impact factor: 19.103
Authors: M Infantino; F Meacci; V Grossi; M Manfredi; F Li Gobbi; P Sarzi-Puttini; F Atzeni; M Benucci Journal: Immunol Res Date: 2017-02 Impact factor: 2.829
Authors: Edward K L Chan; Jan Damoiseaux; Orlando Gabriel Carballo; Karsten Conrad; Wilson de Melo Cruvinel; Paulo Luiz Carvalho Francescantonio; Marvin J Fritzler; Ignacio Garcia-De La Torre; Manfred Herold; Tsuneyo Mimori; Minoru Satoh; Carlos A von Mühlen; Luis E C Andrade Journal: Front Immunol Date: 2015-08-20 Impact factor: 7.561