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Evaluation of a Fully Automated Antinuclear Antibody Indirect Immunofluorescence Assay in Routine Use

Indirect immunofluorescence assay (IFA) using HEp-2 cells as a substrate is the gold standard for detecting antinuclear antibodies (ANA) in patient serum. However, the ANA IFA has labor-intensive nature of the procedure and lacks adequate standardization. To overcome these drawbacks, the automation...

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Autores principales: Choi, Hyun-Woo, Kwon, Yong Jun, Park, Ju-Heon, Lee, Seung-Yeob, Chun, Sejong, Won, Eun Jeong, Lee, Jun Hyung, Choi, Hyun-Jung, Kim, Soo Hyun, Shin, Myung-Geun, Shin, Jong-Hee, Kee, Seung-Jung
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7746920/
https://www.ncbi.nlm.nih.gov/pubmed/33343581
http://dx.doi.org/10.3389/fimmu.2020.607541
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author Choi, Hyun-Woo
Kwon, Yong Jun
Park, Ju-Heon
Lee, Seung-Yeob
Chun, Sejong
Won, Eun Jeong
Lee, Jun Hyung
Choi, Hyun-Jung
Kim, Soo Hyun
Shin, Myung-Geun
Shin, Jong-Hee
Kee, Seung-Jung
author_facet Choi, Hyun-Woo
Kwon, Yong Jun
Park, Ju-Heon
Lee, Seung-Yeob
Chun, Sejong
Won, Eun Jeong
Lee, Jun Hyung
Choi, Hyun-Jung
Kim, Soo Hyun
Shin, Myung-Geun
Shin, Jong-Hee
Kee, Seung-Jung
author_sort Choi, Hyun-Woo
collection PubMed
description Indirect immunofluorescence assay (IFA) using HEp-2 cells as a substrate is the gold standard for detecting antinuclear antibodies (ANA) in patient serum. However, the ANA IFA has labor-intensive nature of the procedure and lacks adequate standardization. To overcome these drawbacks, the automation has been developed and implemented to the clinical laboratory. The purposes of this study were to evaluate the analytical performance of a fully automated Helios ANA IFA analyzer in a real-life laboratory setting, and to compare the time and the cost of ANA IFA testing before and after adopting the Helios system. A total of 3,276 consecutive serum samples were analyzed for ANA using the Helios system from May to August 2019. The positive/negative results, staining patterns, and endpoint titers were compared between Helios and visual readings. Furthermore, the turnaround time and the number of wells used were compared before and after the introduction of Helios system. Of the 3,276 samples tested, 748 were positive and 2,528 were negative based on visual readings. Using visual reading as the reference standard, the overall relative sensitivity, relative specificity, and concordance of Helios reading were 73.3, 99.4, and 93.4% (κ = 0.80), respectively. For pattern recognition, the overall agreement was 70.1% (298/425) for single patterns, and 72.4% (89/123) for mixed patterns. For titration, there was an agreement of 75.9% (211/278) between automated and classical endpoint titers by regarding within ± one titer difference as acceptable. Helios significantly shortened the median turnaround time from 100.6 to 55.7 h (P < 0.0001). Furthermore, routine use of the system reduced the average number of wells used per test from 4 to 1.5. Helios shows good agreement in distinguishing between positive and negative results. However, it still has limitations in positive/negative discrimination, pattern recognition, and endpoint titer prediction, requiring additional validation of results by human observers. Helios provides significant advantages in routine laboratory ANA IFA work in terms of labor, time, and cost savings. We hope that upgrading and developing softwares with more reliable capabilities will allow automated ANA IFA analyzers to be fully integrated into the routine operations of the clinical laboratory.
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spelling pubmed-77469202020-12-19 Evaluation of a Fully Automated Antinuclear Antibody Indirect Immunofluorescence Assay in Routine Use Choi, Hyun-Woo Kwon, Yong Jun Park, Ju-Heon Lee, Seung-Yeob Chun, Sejong Won, Eun Jeong Lee, Jun Hyung Choi, Hyun-Jung Kim, Soo Hyun Shin, Myung-Geun Shin, Jong-Hee Kee, Seung-Jung Front Immunol Immunology Indirect immunofluorescence assay (IFA) using HEp-2 cells as a substrate is the gold standard for detecting antinuclear antibodies (ANA) in patient serum. However, the ANA IFA has labor-intensive nature of the procedure and lacks adequate standardization. To overcome these drawbacks, the automation has been developed and implemented to the clinical laboratory. The purposes of this study were to evaluate the analytical performance of a fully automated Helios ANA IFA analyzer in a real-life laboratory setting, and to compare the time and the cost of ANA IFA testing before and after adopting the Helios system. A total of 3,276 consecutive serum samples were analyzed for ANA using the Helios system from May to August 2019. The positive/negative results, staining patterns, and endpoint titers were compared between Helios and visual readings. Furthermore, the turnaround time and the number of wells used were compared before and after the introduction of Helios system. Of the 3,276 samples tested, 748 were positive and 2,528 were negative based on visual readings. Using visual reading as the reference standard, the overall relative sensitivity, relative specificity, and concordance of Helios reading were 73.3, 99.4, and 93.4% (κ = 0.80), respectively. For pattern recognition, the overall agreement was 70.1% (298/425) for single patterns, and 72.4% (89/123) for mixed patterns. For titration, there was an agreement of 75.9% (211/278) between automated and classical endpoint titers by regarding within ± one titer difference as acceptable. Helios significantly shortened the median turnaround time from 100.6 to 55.7 h (P < 0.0001). Furthermore, routine use of the system reduced the average number of wells used per test from 4 to 1.5. Helios shows good agreement in distinguishing between positive and negative results. However, it still has limitations in positive/negative discrimination, pattern recognition, and endpoint titer prediction, requiring additional validation of results by human observers. Helios provides significant advantages in routine laboratory ANA IFA work in terms of labor, time, and cost savings. We hope that upgrading and developing softwares with more reliable capabilities will allow automated ANA IFA analyzers to be fully integrated into the routine operations of the clinical laboratory. Frontiers Media S.A. 2020-12-04 /pmc/articles/PMC7746920/ /pubmed/33343581 http://dx.doi.org/10.3389/fimmu.2020.607541 Text en Copyright © 2020 Choi, Kwon, Park, Lee, Chun, Won, Lee, Choi, Kim, Shin, Shin and Kee http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Choi, Hyun-Woo
Kwon, Yong Jun
Park, Ju-Heon
Lee, Seung-Yeob
Chun, Sejong
Won, Eun Jeong
Lee, Jun Hyung
Choi, Hyun-Jung
Kim, Soo Hyun
Shin, Myung-Geun
Shin, Jong-Hee
Kee, Seung-Jung
Evaluation of a Fully Automated Antinuclear Antibody Indirect Immunofluorescence Assay in Routine Use
title Evaluation of a Fully Automated Antinuclear Antibody Indirect Immunofluorescence Assay in Routine Use
title_full Evaluation of a Fully Automated Antinuclear Antibody Indirect Immunofluorescence Assay in Routine Use
title_fullStr Evaluation of a Fully Automated Antinuclear Antibody Indirect Immunofluorescence Assay in Routine Use
title_full_unstemmed Evaluation of a Fully Automated Antinuclear Antibody Indirect Immunofluorescence Assay in Routine Use
title_short Evaluation of a Fully Automated Antinuclear Antibody Indirect Immunofluorescence Assay in Routine Use
title_sort evaluation of a fully automated antinuclear antibody indirect immunofluorescence assay in routine use
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7746920/
https://www.ncbi.nlm.nih.gov/pubmed/33343581
http://dx.doi.org/10.3389/fimmu.2020.607541
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