Patients and sampling
Cerebrospinal fluid (CSF) samples were obtained from patients who had been diagnosed with clinically isolated syndrome (CIS), patients with relapsing-remitting multiple sclerosis (RRMS), and individuals in a control group (CG). The control group comprised patients with non-inflammatory central nervous system (CNS) involvement, such as headaches, back pain, and vertigo, for whom lumbar puncture was indicated for differential diagnostic purposes, in order to exclude pathological processes in the CNS.
The patient cohort was subsequently categorized into four distinct groups. The first group, labeled CIS-CIS, consisted of individuals who did not progress to multiple sclerosis (MS) within two years following the collection of cerebrospinal fluid (CSF) samples. The second group, designated CIS-RRMS, encompassed patients who transitioned to relapsing-remitting multiple sclerosis (RRMS) within the same two-year period following CSF collection. The third group, denoted as RRMS, comprised patients for whom marker analysis was conducted at the point of confirmed disease diagnosis. The fourth and final group constituted the control group (CG). Assessment of disability was performed using the Kurtzke Expanded Disability Status Scale (EDSS) by experienced neurologists affiliated with the Department of Neurology at Olomouc University Hospital and Palacký University in Olomouc. Diagnosis was established in accordance with the revised McDonald diagnostic criteria.
Cerebrospinal fluid (CSF) was acquired through a lumbar puncture procedure using an atraumatic needle with patients positioned in a seated posture. The puncture was targeted at the L4/5 intervertebral space. A volume of 10 milliliters of CSF was collected from each patient into a sterile tube devoid of any additives. The sample underwent initial evaluation for cell count and qualitative cytology, followed by centrifugation (at 1100 g for 10 minutes at 4°C) for further biochemical and immunological analysis.