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Relationship Between Polymorphisms of TRPV1 and KCC2 Gene in Children With Febrile Seizures

Completed
Conditions
Febrile Seizure
Interventions
Genetic: Isolated DNA, Real Time PCR
Registration Number
NCT04368936
Lead Sponsor
Institut za Rehabilitaciju Sokobanjska Beograd
Brief Summary

Febrile seizures (FS) are the most common neurological disorder in chilhood. The etiology of FN is still the subject of numerous studies and it is known that it can depend on genetic predisposition.

Detailed Description

Febrile seizures (FS) are the most common neurological disorder in chilhood. It is precisely because of the high incidence of the disease, the age that includes the tendency of repetition, represent a particular challenge in pediatric practice.

FS, as defined by the American Academy of Pediatrics (AAP), are " seizure occurring in febrile children between the ages of 6 and 60 months who do not have an intracranial infection, metabolic disturbance, or history of afebrile seizures ".

Simple febrile seizure is defined as a short (\<15 min) generalized seizure, not repeat within 24 h, that occurs during a febrile illness not resulting from an acute disease of the nervous system in a child aged between 6 months and 5 years, with no neurologic deficits and no previous afebrile seizures. Complex febrile seizures are a focal, or generalized and prolonged seizure, of a duration of greater than 15 min, recurring more than once in 24 h, and/or associated with postictal neurologic abnormalities, more frequently a postictal palsy (Todd's palsy), or with previous neurologic deficits.

The etiology of FN is still the subject of numerous studies and it is known that it can depend on genetic predisposition.

Animal studies have shown that mice without the KCC2 gene have frequent generalized seizures, while those with heterozygous deletion of the KCC2 gene have a reduced threshold for seizure onset. In the human population, mutations of this gene have been reported in children with FN as well as in children with epilepsy. There is no examined an association between polymorphism rs2297201 KCC2 gene and FS.

Studies have shown an association between TRPV1 genes and the appearance of FS in experimental models, however, similar studies in the human population have not been done so far. Studies Moria et al. 2012 showed that polymorphism rs222797 TRPV1 gene involve the regulation of human cortical excitability, glutamate transmission and increased neuronal excitability. The C allele this polymorphism is associated with a greater maximal response to the caspaicin and anadamine agonists. All of this indicate that changes TRPV1 gene that lead to increased channel function may suggest a predisposition for FS.

Since FS are genetically controlled, we want to determine the association of TRPV1 and KCC2 gene polymorphisms with the occurrence of FN.

Recruitment & Eligibility

Status
COMPLETED
Sex
All
Target Recruitment
121
Inclusion Criteria
  • Our research involve patient with diagnosed Febrile Seizure which were hospitalized or recieved ambulatory treatment in University Children´s Hospital in Belgrade
  • For each patient, a diagnosis of FS was made based on the ILAE definition (International Leage
  • The main criterion for inclusion in the study was enhanced FS without prior occurrence of afebrile seizure.
Exclusion Criteria
  • Patients with evidence of intracranial infections and metabolic disbalance
  • Patients with incomplited medical documentation

Study & Design

Study Type
OBSERVATIONAL
Study Design
Not specified
Arm && Interventions
GroupInterventionDescription
CN: Control groupIsolated DNA, Real Time PCRThe control group was made of healthy children older than 5 years of age, which have never had any neurological disorders in their anamnesis and who were patients in preschool or school dispensaries in the city of Belgrade
EFS: group of individuals with Epilepsia and Febrile SeizuresIsolated DNA, Real Time PCRGroup of children with Febrile Seizures, who have developed Epilepsy
SFS : group of individuals with simple FSIsolated DNA, Real Time PCRSimplex febrile seizures (SFS) last shorter than 15 minutes and their type is tonic-clonic. Also, they did not show signs of recidivism during the first 24 hours and were diagnosed at the patients aged from 6th months to 5th year
CFS : group of individuals with complex FSIsolated DNA, Real Time PCRComplex febrile seizures (CFS) were diagnosed at those patients that had focal seizure or epileptic status or seizure having the body temperature lower than 38 degree, which occurred outside of the typical age group and finally which repeated in the first 24 hours again
WFS: group of individuals with FS and without epilepsiaIsolated DNA, Real Time PCRgroup of children with Febrile Seizure and not developed Epilepsia
FS: Febrile SeizuresIsolated DNA, Real Time PCRInvolved patient with diagnosed Febrile Seizures which were hospitalized or recieved ambulatory treatment in University Children´s Hospital in Belgrade. Ages 1-14 years
Primary Outcome Measures
NameTimeMethod
Detection of the polymorphism in the TRPV1 gene polymorphisms2 weeks

The detection of the polymorphism in the TRPV1 gene will be done by the PCR Real time method: During PCR ampification, in addition to primers, an allele of specific oligonucleotide probes is used, which at the 5 'end is labeled with specific fluorescent dye (Reporter dye, eg VIC and FAM), while at the 3' position there is a quencher, which is the role of blocking fluorescence emissions.The fluorescence intensity increases during each cycle and allows us to monitor dynamic reactions in real time.After the final PCR reaction, increasing the fluorescence of the dyes is displayed on the heterozygosity of the test allele. Fluorescence coupling of only one color indicates a homozygous state.VIC dye corresponds to allele C, and FAM dye to allele G.

Detection of the polymorphism in the KCC2 gene polymorphisms2 weeks

The detection of the polymorphism in the KCC2 gene will be done by the PCR Real time method: During PCR ampification, in addition to primers, an allele of specific oligonucleotide probes is used, which at the 5 'end is labeled with specific fluorescent dye (Reporter dye, eg VIC and FAM), while at the 3' position there is a quencher, which is the role of blocking fluorescence emissions.The fluorescence intensity increases during each cycle and allows us to monitor dynamic reactions in real time.After the final PCR reaction, increasing the fluorescence of the dyes is displayed on the heterozygosity of the test allele. Fluorescence coupling of only one color indicates a homozygous state. VIC dye corresponds to allele C, and FAM dye to allele T.

Secondary Outcome Measures
NameTimeMethod
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