Ig G IEF Diagnosing Multiple Sclerosis Multiple Sclerosis
- Slides: 24
Ig. G IEF Diagnosing Multiple Sclerosis
Multiple Sclerosis • CNS disorder • Scar formation on outside of nerve cells of brain and spinal cord • Inflammation destroys covering of nerve cells leaving scar tissue • Nerve cells cannot transmit impulses
Prevalence • Onset at age 15 to 50 years • 70% female • Temperate climate predominant
Symptoms • Optic neuritis • Numbness / weakness in extremities • Instability in walking • Tremors • Loss of bladder control • Heat intolerance • Fatigue
Diagnosis by Exclusion • • Encephalitis CNS Vasculitis Lyme Disease Behet Syndrome Sarcoidosis Syphilis Leukodystrophies • • • Lupus Erythematosus Spastic Paraparesis Vitamin B-12 Deficiency Sjogren’s Syndrome Hereditary Degenerative Disorder • Multifocal Leukoencephalopathy
Criteria for Diagnosis History of two attacks with positive oligoclonal bands or increased Ig. G in CSF
Criteria for Diagnosis History of two attacks with no laboratory abnormality
Criteria for Diagnosis History of two attacks with clinical and paraclinical evidence of lesions, oligoclonal bands, and increased Ig. G in CSF
Criteria for Diagnosis History of two attacks with clinical evidence of at least one lesion, and clinical or paraclinical evidence of another lesion
IEF Theory • Stable p. H gradient with carrier ampholytes in an electrical field
IEF Theory • Stable p. H gradient with carrier ampholytes in an electrical field • Proteins exhibit net + or – charge
IEF Theory • Stable p. H gradient with carrier ampholytes in an electrical field • Proteins exhibit net + or – charge • Migrate toward electrode of opposite charge
IEF Theory • Stable p. H gradient with carrier ampholytes in an electrical field • Proteins exhibit net + or – charge • Migrate toward electrode of opposite charge • Rate of migration ↓ as it reaches isoelectric point
IEF Theory • Stable p. H gradient with carrier ampholytes in an electrical field • Proteins exhibit net + or – charge • Migrate toward electrode of opposite charge • Rate of migration ↓ as it reaches isoelectric point (p. I) • Protein focuses at p. I
IEF Theory • Stable p. H gradient with carrier ampholytes in an electrical field • Proteins exhibit net + or – charge • Migrate toward electrode of opposite charge • Rate of migration ↓ as it reaches isoelectric point (p. I) • Protein focuses at p. I; some drift but regains charge and migrates back
IEF Procedure Step 1: Focus proteins
IEF Procedure Step 2: Transfer proteins to nitrocellulose membrane
IEF Procedure Step 3: React with Ig. G antisera
IEF Procedure Step 4: React with chromagen Step 5: Air dry Total Time: 2. 5 to 3. 0 hours
Interpretation Compare CSF and Serum Looking for intrathecal synthesis indicated by oligoclonal bands in CSF but not in serum
Interpretation Negative – no oligoclonal bands in CSF
Interpretation Mirror Image – same bands in CSF and serum
Interpretation Positive – Oligoclonal bands in CSF but not in serum
Interpretation Positive oligoclonal banding is not diagnosis of MS
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