Clinical Trial Details
— Status: Completed
Administrative data
| NCT number |
NCT05347277 |
| Other study ID # |
2021-6714-19104. |
| Secondary ID |
|
| Status |
Completed |
| Phase |
|
| First received |
|
| Last updated |
|
| Start date |
June 1, 2021 |
| Est. completion date |
August 31, 2021 |
Study information
| Verified date |
April 2022 |
| Source |
Aga Khan University |
| Contact |
n/a |
| Is FDA regulated |
No |
| Health authority |
|
| Study type |
Observational
|
Clinical Trial Summary
Multiple sclerosis (MS) is a demyelinating disease of the central nervous system (CNS) which
commonly leads to disability. The current preferred clinical laboratory test for the
diagnosis is the detection of oligoclonal bands (OCBs) in the cerebrospinal fluid (CSF) by
isoelectric focusing electrophoresis (IEF) followed by immunoblotting.Measuring the levels of
Kappa Free Light Chain (K-FLC) in CSF has been proposed as a potential alternative to the
qualitative assessment of OCBs. The aim of this study is to validate and determine the
diagnostic yield of K-FLC in CSF against OCBs via IEF as gold standard.
Description:
Multiple sclerosis (MS) is a demyelinating disease of the central nervous system (CNS) which
commonly leads to disability. It is the most common disabling neurological disease found in
young adults, most commonly presenting between the ages of 20 to 40 . The current preferred
clinical laboratory test for the diagnosis is the detection of oligoclonal bands (OCBs) in
the cerebrospinal fluid (CSF) by isoelectric focusing electrophoresis (IEF) followed by
immunoblotting. This manual technique requires paired CSF and serum specimens to be run in
parallel, with a subjective visual interpretation. The multi-step method is labor intensive
and costly, with an average time for analytical processing of over 3 hours. There is no
standard definition of OCB amounts required for a clinically positive result. With differing
approaches by varied institutions, positivity can be characterized by anything from 1 to 4
unique CSF bands, which significantly affects sensitivity and specificity of the assay.
Measuring the levels of Kappa Free Light Chain (K-FLC) in CSF has been proposed as a
potential alternative to the qualitative assessment of OCBs. This lab-based validation study
would allow the laboratory to cut spending by reducing technologist time and eliminating
expensive IEF kits. Additionally, K-FLC would replace subjective interpretation with
quantitative values, overcoming the challenges associated with the performance and subjective
interpretation of OCB testing. The aim of this study is to validate and determine the
diagnostic yield of K-FLC in CSF against OCBs via IEF as gold standard.
OCB via IEF is routinely performed at the Section of Chemical Pathology for patients with
suspicion of MS or demyelinating disorders. Paired patient CSF and serum specimens are
collected. The oligoclonal bands are tested by IEF on agarose gel using Interlab diagnostics
CSF IEF kit and manual immunoblotting steps to transfer the proteins on transfer membranes.
Simultaneous analysis is also conducted on serum samples. The pattern of oligoclonal bands in
CSF is reviewed by pathologist and their findings are noted as positive or negative for
presence or absence of CSF OCB or matching, that is, bands present in both serum and CSF.
The samples received during the last 3 months from June to August for OCBS via IEF are saved
in the lab in frozen conditions, stable for analysis. 0.5 ml CSF sample will be utilized for
analysis by nephelometry on Beckman Immage-800 analyzer using kits from Freelite, The Binding
Site, according to the manufacturer's instructions.
To ensure confidentiality, all identification details will be anonymized, and study
identifiers will be allotted. Additionally, all data will be stored in a password protected
electronic format by the PI.
Keeping in view the low anticipated frequency of demyelinating disorders at a single center;
and to achieve significant power and reliability of statistical analysis, a minimum sample
size of 60 CSF samples for K-FLC analysis will be included with stratification into 3 groups:
Positive (n=20), negative (n=20) and matching (n=20), for OCBs.
Data analysis will be done on SPSS version 25. Shapiro Wilk test will be used to check
normality of data. Diagnostic accuracy of CSF K-FLC will be calculated on the basis of
sensitivity, specificity, PPV and NPV taking OCB as gold standard. For further analysis,
Receiver operative characteristic (ROC) curve will be plotted for OCB and area under the
curve (AUC) will be calculated and optimal cut-off will be generated using Youden's Index.
For the validation agreement between CSF L-FLC and IEF will be assessed by Cohen's kappa
statistics.
