CSF Protein Determination Flashcards
2 types of method in Total Protein
Turbidimetric Method
Dye-Binding Method
preferred method, precipitates both albumin and globulin
Trichloroacetic acid (3% TCA)
precipitates albumin only
▪ To precipitate globulins, we add sodium sulfate
Sulfosalicylic acid (3% SSA)
Dye-Binding Method:
Coomasie Brilliant Blue
protein binds to dye → dye turns red to blue (intense blue color = high protein present)
assess the integrity of blood brain barrier
CSF/Serum Albumin index
To compute:
o Normal value:
o Abnormal:
To compute: CSF Albumin (mg/dL) / Serum Albumin (g/dL)
o Normal value: <9 = intact BBB
o Abnormal: >9
Slight impairment
9-14
Moderate impairment
15-30
Severe impairment
> 30
Complete damage to BBB
100
assess conditions the IgG production within the CNS (ie. Multiple sclerosis – increase plasma cells)
IgG Index
IgG Index Value
Normal:
Abnormal:
Normal: <0.70
Abnormal: >0.70 (there’s an increase IgG production → Multiple sclerosis)
Presence of 2 oligoclonal bands in CSF and not in serum
o Multiple sclerosis
o Neurosyphilis
o Encephalitis
o Neoplastic disorder
o Guillain-Barre syndrome
Detection of oligoclonal bands in the gamma region → immunoglobulin production
CSF Electrophoresis
- Demyelinating disorder
- Production of antibodies against myelin sheath
Multiple Sclerosis
Multiple Sclerosis Findings:
o (+) anti-myelin sheath autoantibody
o (+) oligoclonal band in CSF but not in serum
o (+) Myelin Basic protein (MBP) – protein component of myelin sheath, monitors MS
o Increased IgG index
CSF Glucose
Normal Value:
CSF Glucose
Normal Value:
* 60-70% of blood glucose (65%, 2/3) *accd. Strasinger
* 50-80 mg/dL *accd. Henry’s
Measurement of glucose in CSF is always performed in conjunction with ____ glucose
blood
Specimen for blood glucose should be collected _____ before spinal tap to allow equilibration of CSF and plasma glucose
2 hours
Increased CSF Glucose
Diabetes mellitus
Decreased CSF Glucose
- Bacterial (Decreased CSF glucose), tubercular, and fungal meningitis
- Alterations in glucose transport across BBB
- Increased used of glucose by brain cells
Normal CSF glucose
Viral meningitis
waste product of glucose metabolism, inversely proportional with glucose
Lactate
Increased CSF Lactate
- Bacterial meningitis (bacteria utilized glucose → lactate >35mg/dL)
- Tubercular and fungal meningitis (>25 mg/dL)
- Hypoxia (tissues are destroyed causing release of lactate)
CSF Lactate
Normal values:
CSF Lactate
Normal values: 10-24 mg/dL
Normal CSF lactate (<25 mg/dL)
Viral meningitis
A product of ammonia (NH3) and a-ketoglutarate in braincells
CSF Glutamine
Indirect test for the presence of excess NH3 in CSF
(High NH3 = High Glutamine)
CSF Glutamine
Normal value:
CSF Glutamine
Normal value: 8-18 mg/dL
Increased CSF Glutamine
- Disturbance of consciousness (>35 mg/dL)
- Reye’s syndrome – 75% of patient with Reye’s syndrome have increased CSF glutamine
found in brain tissue
LD1 and 2
Lymphocytes
LD2 and 3
Neutrophils
LD4 and 5
Increased Creatinine Kinase
Stroke, multiple sclerosis, degenerative disorders, brain tumors, viral and bacterial meningitis, epileptic seizure
Increased Aspartate Aminotransferase
Intracerebral and subarachnoid hemorrhage, bacterial meningitis
bacterial meningitis → high neutrophils
5>4>3>2>1
If the distribution of LD enzyme in serum is seen on CSF (predominant LD2)
neurologic abnormality
Normal distribution of LD enzyme in serum
LD2>LD1>LD3>LD4>LD5
Normal distribution of LD enzyme in CSF
LD1>LD2>LD3>LD4>LD5
If the distribution of LD enzyme in CSF is seen on serum (predominant LD1) → flipped pattern
Acute Myocardial Infarction or Hemolytic anemia
Common agents: Group B streptococci
Neonates
Common agents: E. coli and other gram negative bacilli
Newborn to 1 month
Common agents: Neisseria meningitis
Children (>3 months )
Common agents: Streptococcus pneumoniae
Children (>3 months )
Common agents: Haemophilus influenzae
3 month to 18 years old
Common agents: Listeria monocytogenes
Newborns, Adults, alcoholics, immunocompromised
Laboratory Findings:
* WBC –
* Protein –
* Glucose –
* Lactate -
* Others
o (?) Gram stain
o (?) Culture
o (?) Limulus Lysate test
Laboratory Findings:
* WBC – Increased neutrophils
* Protein – Increased
* Glucose – Decreased
* Lactate - Increased
* Others
o (+) Gram stain
o (+) Culture
o (+) Limulus Lysate test
Detects gram negative bacterial endotoxin in body fluids and surgical instruments
Limulus Lysate test
Limulus Lysate test Reagent:
Blood of horseshoe crabs (Limulus polyhemus)
Blue in color because of hemocyanin that contains copper
Endotoxin → Amoebocyte → release lysate →
Positive reaction: Clot formation
Blood also contains WBCS called
amoebocyte
Spirochetal Meningitis
Neurosyphilis – caused by
Neuroborreliosis – caused by
Neurosyphilis – caused by T. pallidum
Neuroborreliosis – caused by B. burgdorfer
Spirochetal Meningitis
Diagnosis:
Diagnosis:
* Nontreponemal tests (VDRL, FTA-Abs)
* ELISA
* Western blot
recommended test of CDC for neurosyphilis
VDRL
Tubercular Meningitis
Agent:
Tubercular Meningitis
Agent: Mycobacterium tuberculosis
Tubercular Meningitis
Laboratory findings:
* WBC –
* Protein –
* Glucose –
* Lactate –
* (?) AFB stain
* (?) Pellicle/web-like clot formation after stored/stand 12-24 hours in refrigerator
Tubercular Meningitis
Laboratory findings:
* WBC – Increased lymphocyte and monocyte
* Protein – increased
* Glucose – Decreased
* Lactate – Increased
* (+) AFB stain
* (+) Pellicle/web-like clot formation after stored/stand 12-24 hours in refrigerator
Fungal Meningitis
Agent:
Fungal Meningitis
Agent: Cryptococcus neoformans *most frequently isolated pathogen in the CSF
Fungal Meningitis
Laboratory findings:
* WBC –
* Protein –
* Glucose –
* Lactate –
* (?) Gram stain – starburst pattern
* (?) India Ink (Negative/Indirect stain – background only)
* (?) Latex agglutinations test – detects the presence of antigen of C. neoformans
Fungal Meningitis
Laboratory findings:
* WBC – Increase lymphocyte and monocyte
* Protein – Increased
* Glucose – Decreased
* Lactate – Increased
* (+) Gram stain – starburst pattern
* (+) India Ink (Negative/Indirect stain – background only)
* (+) Latex agglutinations test – detects the presence of antigen of C. neoformans
Amoebic Meningoencephalitis
Agents:
* Naegleria fowleri –
* Acanthamoeba species and Balamuthia mandrillaris –
Amoebic Meningoencephalitis
Agents:
* Naegleria fowleri – Primary amoebic meningoencephalitis
* Acanthamoeba species and Balamuthia mandrillaris – causes Granulomatous amoebic encephalitis
Amoebic Meningoencephalitis
Laboratory Findings:
* WBC –
* Protein –
* Glucose -
* Lactate –
* (?) RBCs
* (?) Acridine orange stain
Laboratory Findings:
* WBC – Increased neutrophils
* Protein – Increased
* Glucose - Decreased
* Lactate – Increased
* (+) RBCs
* (+) Acridine orange stain
agents can be identified with Wright-stained
smear but macrophage can be mistakenly identified as protozoa
Acridine orange stain
Amoeba
brick red
Viral Meningitis
Agents:
Viral Meningitis
Agents: Enteroviruses (Coxsackievirus, Echovirus, Poliovirus), Arbovirus
WBC
bright green
Viral Meningitis
Laboratory Findings:
* WBC –
* Protein -
* Glucose -
* Lactate -
* RT-PCR
Viral Meningitis
Laboratory Findings:
* WBC – Increased lymphocytes
* Protein - Increased
* Glucose - Normal
* Lactate - Normal
* RT-PCR
gold standard for diagnosis of viral meningitis
RT-PCR
