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.