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Diagnosing Multiple Sclerosis
IgG IEF Diagnosing Multiple Sclerosis
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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
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Prevalence Onset at age 15 to 50 years 70% female
Temperate climate predominant
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Symptoms Optic neuritis Numbness / weakness in extremities
Instability in walking Tremors Loss of bladder control Heat intolerance Fatigue
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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
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Criteria for Diagnosis
History of two attacks with positive oligoclonal bands or increased IgG in CSF Probable MS
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Criteria for Diagnosis
History of two attacks with no laboratory abnormality Clinically Probable MS
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Criteria for Diagnosis
History of two attacks with clinical and paraclinical evidence of lesions, oligoclonal bands, and increased IgG in CSF Laboratory-Supported Definitive MS
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Criteria for Diagnosis
History of two attacks with clinical evidence of at least one lesion, and clinical or paraclinical evidence of another lesion Clinically Definitive MS
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IEF Theory Stable pH gradient with carrier ampholytes in an electrical field
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IEF Theory Stable pH gradient with carrier ampholytes in an electrical field Proteins exhibit net + or – charge
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IEF Theory Stable pH gradient with carrier ampholytes in an electrical field Proteins exhibit net + or – charge Migrate toward electrode of opposite charge
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IEF Theory Stable pH 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
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IEF Theory Stable pH 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 (pI) Protein focuses at pI
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IEF Theory Stable pH 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 (pI) Protein focuses at pI; some drift but regains charge and migrates back
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IEF Procedure Step 1: Focus proteins
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IEF Procedure Step 2: Transfer proteins to nitrocellulose membrane
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IEF Procedure Step 3: React with IgG antisera
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IEF Procedure Step 4: React with chromagen Step 5: Air dry
Total Time: 2.5 to 3.0 hours
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Looking for intrathecal synthesis oligoclonal bands in CSF
Interpretation Compare CSF and Serum Looking for intrathecal synthesis indicated by oligoclonal bands in CSF but not in serum
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Interpretation Negative – no oligoclonal bands in CSF
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Interpretation Mirror Image – same bands in CSF and serum
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Interpretation Positive – Oligoclonal bands in CSF but not in serum
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Interpretation Positive oligoclonal banding is not diagnosis of MS
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