A second-generation ELISA (STRATIFY JCV™ DxSelect™) for detection of JC virus antibodies in human serum and plasma to support progressive multifocal leukoencephalopathy risk stratification
Section snippets
Background
Progressive multifocal leukoencephalopathy (PML) is a rare demyelinating disease of the central nervous system associated with the use of certain immunomodulatory agents.1 While multiple host and viral factors contribute to the development of PML, infection with a common polyomavirus, JC virus (JCV), is a known prerequisite.2 We previously reported the development and validation of a novel two-step enzyme-linked immunosorbent assay (ELISA) to detect JCV antibodies in human serum or plasma and
Objectives
Our objectives were to enhance the robustness and performance characteristics of the two-step JCV antibody ELISA, translate it into a kit format, validate the enhanced assay, and demonstrate its potential utility as a tool for stratifying multiple sclerosis (MS) patients by risk of developing PML.
Samples
Assay cut-points were determined using serum samples from 228 urinary JCV DNA-positive patients (a subset of the 875 MS patients enrolled in the Safety of Tysabri Re-dosing and Treatment [STRATA] study [NCT00297232] whose data were used for development of the original two-step JCV antibody assay)3 and serum samples from 1091 MS patients collected at baseline in the STRATIFY-1 study (NCT01070823). Serum samples from 792 MS patients enrolled in the STRATIFY-2 study (NCT01070836) and 538 patients
ELISA plate surface chemistry and coating buffer selection
Signal-to-noise ratios were calculated for both positive control 1 (PC1) and positive control 2 (PC2) relative to NC for all surface chemistries and coating buffer conditions, with the PC2/NC ratio being the primary parameter, as it reflects the signal-to-noise ratio of low levels of JCV antibodies. MediSorp and MaxiSorp plates with Focus's proprietary PBS yielded the two highest PC2/NC ratios, which were 3.8 and 3.6, respectively (Supplementary Table A). MediSorp plates with Focus's
Discussion
We previously described the development and validation of a two-step JCV antibody ELISA. Here, we reported the development of a second-generation JCV antibody ELISA that offered two major improvements. First, stabilization of JC VLP onto the microtiter plates and the development of a ready-to-use kit improved the reproducibility and ease of use of the assay. Immobilization of JC VLP on the plates minimized variability resulting from dilution and coating of JC VLP. Second, the dynamic range of
Funding
Funding for the assay development and validation studies was provided by Biogen Idec Inc.
Competing interests
Peter Lee, Albert Castro, Dipeshkumar Jaiswal, and Suzanne Rivas are currently employed by Focus Diagnostics. Tatiana Plavina, Melissa Berman, Brian Schlain, and Meena Subramanyam are currently employed by and own stock in Biogen Idec.
Ethical approval
Not required.
Acknowledgments
The authors thank the laboratory staff at Focus Diagnostics, Dr. Vijaya Nagabhushanam and the laboratory staff at National Jewish Health, and Dr. Nang Nguyen and the laboratory staff at University of Rochester Medical Center for performing the validation experiments. The authors also thank Dr. Wayne Hogrefe for scientific discussions on assay development. Editorial support was provided by Joshua Safran of Infusion Communications and was funded by Biogen Idec Inc. and Elan Pharmaceuticals, Inc.
References (9)
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Multi-site analytical validation of an assay to detect anti-JCV antibodies in human serum and plasma
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Anti-JC virus antibodies: implications for PML risk stratification
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2020, Infectious Disease Clinics of North AmericaCitation Excerpt :Multiple biologics have been associated with the development of JCPyV encephalopathy but screening has been validated only with natalizumab because the observed incidence is high enough to permit estimated risk stratification in this patient population.70,114 The screening algorithm should be used in conjunction with risk estimates based on anti-JCPyV IgG antibody index serostatus, prior immunosuppression, and duration of natalizumab exposure.84,110,112–114 After natalizumab initiation, patients should be monitored closely for development of new neurologic deficits.84,113
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2019, Journal of the Neurological SciencesCitation Excerpt :To stratify PML risk in natalizumab-treated patients, Biogen developed an analytically validated enzyme-linked immunosorbent assay (the STRATIFY JCV™ assay, Focus Diagnostics, Cypress, CA, USA) to detect the presence of JCV antibodies in serum [15,16]. An enhanced assay (STRATIFY JCV DxSelect™) in a new, easier-to-use kit format has been available since March 2012 [17]. This test has improved sensitivity to facilitate detection of anti-JCV antibody positive patients with low JCV antibody levels [17].
- c
Co-lead authors who contributed equally to this work.