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Novel Automation of an Enzyme-Linked Immunosorbent Spot Assay Testing Method: Comparable Diagnostic Performance of the T-SPOT.TB Test Using Manual Density Gradient Cell Isolation versus Automated Positive Selection with the T-Cell Select Kit
The diagnosis of latent tuberculosis (TB) infection (LTBI) is critical to improve TB treatment and control, and the T-SPOT.TB test is a commercial enzyme-linked immunosorbent spot assay used for this purpose. The objective of the study was to increase automation and extend the time between blood col...
Autores principales: | , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Microbiology
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9491199/ https://www.ncbi.nlm.nih.gov/pubmed/36040150 http://dx.doi.org/10.1128/jcm.00551-22 |
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author | Wang, Shu-Hua Rajaram, Murgesean V. S. Trollip, Andre Wu, Qian Ayala, Doris Garza, Danyelle Benavidez, Marielena A. Fox, Kelsie Aguillón-Durán, Genesis P. Vargas-Orozco, Erika A. Torres, Lou Yu, Lianbo Ahmed, S. Rumel Aspden, Megan Jackson-Soutter, Thomas Coxon, Catherine Brignall, Ruth Restrepo, Blanca I. |
author_facet | Wang, Shu-Hua Rajaram, Murgesean V. S. Trollip, Andre Wu, Qian Ayala, Doris Garza, Danyelle Benavidez, Marielena A. Fox, Kelsie Aguillón-Durán, Genesis P. Vargas-Orozco, Erika A. Torres, Lou Yu, Lianbo Ahmed, S. Rumel Aspden, Megan Jackson-Soutter, Thomas Coxon, Catherine Brignall, Ruth Restrepo, Blanca I. |
author_sort | Wang, Shu-Hua |
collection | PubMed |
description | The diagnosis of latent tuberculosis (TB) infection (LTBI) is critical to improve TB treatment and control, and the T-SPOT.TB test is a commercial enzyme-linked immunosorbent spot assay used for this purpose. The objective of the study was to increase automation and extend the time between blood collection and processing for the T-SPOT.TB test from 0 to 8 h to 0 to 54 h. The previous maximum time between blood collection and processing for the T-SPOT.TB test is 32 h using T-Cell Xtend. For this, we compared the T-SPOT.TB test using manual peripheral blood mononuclear cell (PBMC) isolation by density gradient separation at 0 to 8 h (reference method, control arm) to an automated PBMC isolation method using magnetic beads (T-Cell Select kit) at 0 to 55 h postcollection. A total of 620 subjects were enrolled from 4 study sites, and blood samples were collected from each volunteer, comprising 1,850 paired samples in total. Overall agreement between both methods was 96.8% (confidence interval [CI], 95.9 to 97.6%), with 95.8% (CI, 93.5 to 97.5%) positive and 97.1% negative agreement (CI, 96.1 to 97.9%). In summary, there was a strong overall agreement between the automated and manual T-SPOT.TB test processing methods. The results suggest that the T-SPOT.TB test can be processed using automated positive selection with magnetic beads using T-Cell Select to decrease hands-on time. Also, this cell isolation method allowed for the time between blood collection and processing to range from 0 to 55 h. Additional studies in larger and diverse patient populations including immunocompromised and pediatric patients are needed. |
format | Online Article Text |
id | pubmed-9491199 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Society for Microbiology |
record_format | MEDLINE/PubMed |
spelling | pubmed-94911992022-09-22 Novel Automation of an Enzyme-Linked Immunosorbent Spot Assay Testing Method: Comparable Diagnostic Performance of the T-SPOT.TB Test Using Manual Density Gradient Cell Isolation versus Automated Positive Selection with the T-Cell Select Kit Wang, Shu-Hua Rajaram, Murgesean V. S. Trollip, Andre Wu, Qian Ayala, Doris Garza, Danyelle Benavidez, Marielena A. Fox, Kelsie Aguillón-Durán, Genesis P. Vargas-Orozco, Erika A. Torres, Lou Yu, Lianbo Ahmed, S. Rumel Aspden, Megan Jackson-Soutter, Thomas Coxon, Catherine Brignall, Ruth Restrepo, Blanca I. J Clin Microbiol Immunoassays The diagnosis of latent tuberculosis (TB) infection (LTBI) is critical to improve TB treatment and control, and the T-SPOT.TB test is a commercial enzyme-linked immunosorbent spot assay used for this purpose. The objective of the study was to increase automation and extend the time between blood collection and processing for the T-SPOT.TB test from 0 to 8 h to 0 to 54 h. The previous maximum time between blood collection and processing for the T-SPOT.TB test is 32 h using T-Cell Xtend. For this, we compared the T-SPOT.TB test using manual peripheral blood mononuclear cell (PBMC) isolation by density gradient separation at 0 to 8 h (reference method, control arm) to an automated PBMC isolation method using magnetic beads (T-Cell Select kit) at 0 to 55 h postcollection. A total of 620 subjects were enrolled from 4 study sites, and blood samples were collected from each volunteer, comprising 1,850 paired samples in total. Overall agreement between both methods was 96.8% (confidence interval [CI], 95.9 to 97.6%), with 95.8% (CI, 93.5 to 97.5%) positive and 97.1% negative agreement (CI, 96.1 to 97.9%). In summary, there was a strong overall agreement between the automated and manual T-SPOT.TB test processing methods. The results suggest that the T-SPOT.TB test can be processed using automated positive selection with magnetic beads using T-Cell Select to decrease hands-on time. Also, this cell isolation method allowed for the time between blood collection and processing to range from 0 to 55 h. Additional studies in larger and diverse patient populations including immunocompromised and pediatric patients are needed. American Society for Microbiology 2022-08-30 /pmc/articles/PMC9491199/ /pubmed/36040150 http://dx.doi.org/10.1128/jcm.00551-22 Text en Copyright © 2022 Wang et al. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Immunoassays Wang, Shu-Hua Rajaram, Murgesean V. S. Trollip, Andre Wu, Qian Ayala, Doris Garza, Danyelle Benavidez, Marielena A. Fox, Kelsie Aguillón-Durán, Genesis P. Vargas-Orozco, Erika A. Torres, Lou Yu, Lianbo Ahmed, S. Rumel Aspden, Megan Jackson-Soutter, Thomas Coxon, Catherine Brignall, Ruth Restrepo, Blanca I. Novel Automation of an Enzyme-Linked Immunosorbent Spot Assay Testing Method: Comparable Diagnostic Performance of the T-SPOT.TB Test Using Manual Density Gradient Cell Isolation versus Automated Positive Selection with the T-Cell Select Kit |
title | Novel Automation of an Enzyme-Linked Immunosorbent Spot Assay Testing Method: Comparable Diagnostic Performance of the T-SPOT.TB Test Using Manual Density Gradient Cell Isolation versus Automated Positive Selection with the T-Cell Select Kit |
title_full | Novel Automation of an Enzyme-Linked Immunosorbent Spot Assay Testing Method: Comparable Diagnostic Performance of the T-SPOT.TB Test Using Manual Density Gradient Cell Isolation versus Automated Positive Selection with the T-Cell Select Kit |
title_fullStr | Novel Automation of an Enzyme-Linked Immunosorbent Spot Assay Testing Method: Comparable Diagnostic Performance of the T-SPOT.TB Test Using Manual Density Gradient Cell Isolation versus Automated Positive Selection with the T-Cell Select Kit |
title_full_unstemmed | Novel Automation of an Enzyme-Linked Immunosorbent Spot Assay Testing Method: Comparable Diagnostic Performance of the T-SPOT.TB Test Using Manual Density Gradient Cell Isolation versus Automated Positive Selection with the T-Cell Select Kit |
title_short | Novel Automation of an Enzyme-Linked Immunosorbent Spot Assay Testing Method: Comparable Diagnostic Performance of the T-SPOT.TB Test Using Manual Density Gradient Cell Isolation versus Automated Positive Selection with the T-Cell Select Kit |
title_sort | novel automation of an enzyme-linked immunosorbent spot assay testing method: comparable diagnostic performance of the t-spot.tb test using manual density gradient cell isolation versus automated positive selection with the t-cell select kit |
topic | Immunoassays |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9491199/ https://www.ncbi.nlm.nih.gov/pubmed/36040150 http://dx.doi.org/10.1128/jcm.00551-22 |
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