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Consequences of the Edge Effect in a Commercial Enzyme-Linked Immunosorbent Assay for the Diagnosis of Lyme Neuroborreliosis

The diagnosis of Lyme neuroborreliosis (LNB) is based on neurological symptoms, cerebrospinal fluid (CSF) pleocytosis, and intrathecally produced Borrelia-specific antibodies. In most cases, the presence of intrathecally produced Borrelia-specific antibodies is determined by using an enzyme-linked i...

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Autores principales: van Gorkom, Tamara, van Arkel, Gijs H. J., Voet, Willem, Thijsen, Steven F. T., Kremer, Kristin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8288263/
https://www.ncbi.nlm.nih.gov/pubmed/33980651
http://dx.doi.org/10.1128/JCM.03280-20
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author van Gorkom, Tamara
van Arkel, Gijs H. J.
Voet, Willem
Thijsen, Steven F. T.
Kremer, Kristin
author_facet van Gorkom, Tamara
van Arkel, Gijs H. J.
Voet, Willem
Thijsen, Steven F. T.
Kremer, Kristin
author_sort van Gorkom, Tamara
collection PubMed
description The diagnosis of Lyme neuroborreliosis (LNB) is based on neurological symptoms, cerebrospinal fluid (CSF) pleocytosis, and intrathecally produced Borrelia-specific antibodies. In most cases, the presence of intrathecally produced Borrelia-specific antibodies is determined by using an enzyme-linked immunosorbent assay (ELISA). The edge effect is a known phenomenon in ELISAs and can negatively influence the assay reproducibility and repeatability, as well as index calculations of sample pairs which are tested in the same run. For LNB diagnostics, an index calculation is used for which the relative amounts of Borrelia-specific antibodies in CSF and serum are measured to calculate a CSF/serum quotient, which is needed to calculate the Borrelia-specific antibody index (AI). The presence of an edge effect in an ELISA used for LNB diagnostics may thus have implications. In this study, we investigated the intra-assay variation of the commercial Enzygnost Lyme link VlsE/IgG ELISA used for LNB diagnostics and showed the presence of an edge effect. Minor adaptations in the ELISA protocol decreased this effect. The adapted protocol was subsequently used to test 149 CSF-serum pairs of consecutive patients received in a routine diagnostic laboratory. By simulation, we showed that, if the standard protocol would have been used, then the edge effect for this study population could have resulted in 15 (10.1%) false-pathological and two (1.3%) false-normal Borrelia-specific IgG AIs. Thus, the observed edge effect can lead to inaccurate LNB diagnoses. Our study underlines that the edge effect should be investigated when ELISAs are implemented in routine diagnostics, as this phenomenon can occur in any ELISA.
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spelling pubmed-82882632022-01-19 Consequences of the Edge Effect in a Commercial Enzyme-Linked Immunosorbent Assay for the Diagnosis of Lyme Neuroborreliosis van Gorkom, Tamara van Arkel, Gijs H. J. Voet, Willem Thijsen, Steven F. T. Kremer, Kristin J Clin Microbiol Immunoassays The diagnosis of Lyme neuroborreliosis (LNB) is based on neurological symptoms, cerebrospinal fluid (CSF) pleocytosis, and intrathecally produced Borrelia-specific antibodies. In most cases, the presence of intrathecally produced Borrelia-specific antibodies is determined by using an enzyme-linked immunosorbent assay (ELISA). The edge effect is a known phenomenon in ELISAs and can negatively influence the assay reproducibility and repeatability, as well as index calculations of sample pairs which are tested in the same run. For LNB diagnostics, an index calculation is used for which the relative amounts of Borrelia-specific antibodies in CSF and serum are measured to calculate a CSF/serum quotient, which is needed to calculate the Borrelia-specific antibody index (AI). The presence of an edge effect in an ELISA used for LNB diagnostics may thus have implications. In this study, we investigated the intra-assay variation of the commercial Enzygnost Lyme link VlsE/IgG ELISA used for LNB diagnostics and showed the presence of an edge effect. Minor adaptations in the ELISA protocol decreased this effect. The adapted protocol was subsequently used to test 149 CSF-serum pairs of consecutive patients received in a routine diagnostic laboratory. By simulation, we showed that, if the standard protocol would have been used, then the edge effect for this study population could have resulted in 15 (10.1%) false-pathological and two (1.3%) false-normal Borrelia-specific IgG AIs. Thus, the observed edge effect can lead to inaccurate LNB diagnoses. Our study underlines that the edge effect should be investigated when ELISAs are implemented in routine diagnostics, as this phenomenon can occur in any ELISA. American Society for Microbiology 2021-07-19 /pmc/articles/PMC8288263/ /pubmed/33980651 http://dx.doi.org/10.1128/JCM.03280-20 Text en Copyright © 2021 van Gorkom 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
van Gorkom, Tamara
van Arkel, Gijs H. J.
Voet, Willem
Thijsen, Steven F. T.
Kremer, Kristin
Consequences of the Edge Effect in a Commercial Enzyme-Linked Immunosorbent Assay for the Diagnosis of Lyme Neuroborreliosis
title Consequences of the Edge Effect in a Commercial Enzyme-Linked Immunosorbent Assay for the Diagnosis of Lyme Neuroborreliosis
title_full Consequences of the Edge Effect in a Commercial Enzyme-Linked Immunosorbent Assay for the Diagnosis of Lyme Neuroborreliosis
title_fullStr Consequences of the Edge Effect in a Commercial Enzyme-Linked Immunosorbent Assay for the Diagnosis of Lyme Neuroborreliosis
title_full_unstemmed Consequences of the Edge Effect in a Commercial Enzyme-Linked Immunosorbent Assay for the Diagnosis of Lyme Neuroborreliosis
title_short Consequences of the Edge Effect in a Commercial Enzyme-Linked Immunosorbent Assay for the Diagnosis of Lyme Neuroborreliosis
title_sort consequences of the edge effect in a commercial enzyme-linked immunosorbent assay for the diagnosis of lyme neuroborreliosis
topic Immunoassays
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8288263/
https://www.ncbi.nlm.nih.gov/pubmed/33980651
http://dx.doi.org/10.1128/JCM.03280-20
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