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IgG IEF PowerPoint Presentation

IgG IEF

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IgG IEF

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  1. IgG IEF Diagnosing Multiple Sclerosis

  2. 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

  3. Prevalence • Onset at age 15 to 50 years • 70% female • Temperate climate predominant

  4. Symptoms • Optic neuritis • Numbness / weakness in extremities • Instability in walking • Tremors • Loss of bladder control • Heat intolerance • Fatigue

  5. 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 Diagnosis by Exclusion

  6. Criteria for Diagnosis History of two attacks with positive oligoclonal bands or increased IgG in CSF Probable MS

  7. Criteria for Diagnosis History of two attacks with no laboratory abnormality Clinically Probable MS

  8. 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

  9. 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

  10. IEF Theory • Stable pH gradient with carrier ampholytes in an electrical field

  11. IEF Theory • Stable pH gradient with carrier ampholytes in an electrical field • Proteins exhibit net + or – charge

  12. IEF Theory • Stable pH gradient with carrier ampholytes in an electrical field • Proteins exhibit net + or – charge • Migrate toward electrode of opposite charge

  13. 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

  14. 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

  15. 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

  16. IEF Procedure Step 1: Focus proteins

  17. IEF Procedure Step 2: Transfer proteins to nitrocellulose membrane

  18. IEF Procedure Step 3: React with IgG antisera

  19. IEF Procedure Step 4: React with chromagen Step 5: Air dry Total Time: 2.5 to 3.0 hours

  20. Interpretation Compare CSF and Serum Looking for intrathecal synthesis indicated by oligoclonal bands in CSF but not in serum

  21. Interpretation Negative – no oligoclonal bands in CSF

  22. Interpretation Mirror Image – same bands in CSF and serum

  23. Interpretation Positive – Oligoclonal bands in CSF but not in serum

  24. Interpretation Positive oligoclonal banding is not diagnosis of MS