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Lumbar Puncture Reference Guide

Purpose

This reference provides comprehensive CSF study recommendations organized by: 1. Core studies - Always order for any diagnostic LP 2. Diagnosis-specific panels - Tailored to clinical suspicion 3. Advanced testing - Newer molecular and genomic options 4. Special procedures - Therapeutic LPs (NPH, IIH)


CORE STUDIES (Always Order)

These studies should be sent with every diagnostic LP to maximize yield from a single procedure.

Study Tube Volume Rationale Normal Values
Opening pressure - - Elevated in IIH, meningitis, mass effect 10-20 cm H2O
Cell count with differential Tube 1 AND Tube 4 1-2 mL each Tube 4 corrects for traumatic tap WBC <5, RBC 0
Protein Tube 2 1 mL Elevated in infection, GBS, tumor, inflammation 15-45 mg/dL
Glucose Tube 2 1 mL Low in bacterial/fungal/carcinomatous meningitis 50-80 mg/dL (>60% serum)
Gram stain Tube 3 1 mL Rapid bacterial identification No organisms
Bacterial culture Tube 3 2-3 mL Gold standard for bacterial meningitis No growth

CRITICAL: Always send serum glucose at time of LP to calculate CSF:serum ratio.

Tube order matters: Tubes 1-4 should be collected and labeled sequentially. Cell count on both Tube 1 and Tube 4 helps distinguish traumatic tap (RBCs decrease tube 1→4) from true hemorrhage (RBCs stable).


DIAGNOSIS-SPECIFIC PANELS

Meningitis/Encephalitis Panel

Study Priority Indication Turnaround Notes
BioFire FilmArray ME Panel URGENT Suspected bacterial/viral/fungal meningitis ~1 hour 14 pathogens; 94% sensitivity, 99.8% specificity
HSV-1/2 PCR URGENT Encephalitis, temporal lobe involvement 24-48h Most common treatable cause of encephalitis
VZV PCR URGENT Zoster history, dermatomal symptoms 24-48h Can cause vasculopathy
Enterovirus PCR URGENT Summer/fall, aseptic meningitis 24-48h Most common viral meningitis
Cryptococcal antigen URGENT Immunocompromised, HIV 1-2h More sensitive than India ink
AFB smear and culture ROUTINE TB risk factors, chronic meningitis Smear 24h; culture 6-8 weeks Low sensitivity; consider TB PCR
Fungal culture ROUTINE Immunocompromised, endemic areas 4-6 weeks Hold for extended incubation

BioFire FilmArray ME Panel Targets (14 pathogens):

Bacteria Viruses Yeast
E. coli K1 HSV-1, HSV-2 Cryptococcus neoformans/gattii
H. influenzae VZV
L. monocytogenes CMV
N. meningitidis Enterovirus
S. pneumoniae HHV-6
S. agalactiae Human parechovirus

Multiple Sclerosis / Demyelinating Disease Panel

Study Priority Indication Target Finding
Oligoclonal bands (CSF + serum) ROUTINE MS diagnosis, CIS evaluation ≥2 CSF-specific bands
IgG index ROUTINE Intrathecal antibody synthesis >0.7 elevated
Myelin basic protein ROUTINE Acute demyelination Elevated during relapse
CSF cytology ROUTINE Rule out lymphoma/carcinomatous Negative
NMO-IgG (AQP4-Ab) ROUTINE Longitudinal myelitis, severe ON Positive → NMOSD
MOG-IgG ROUTINE Atypical features, ADEM-like Positive → MOGAD

Autoimmune/Inflammatory Encephalitis Panel

Study Priority Indication Target Finding
Autoimmune encephalitis panel (Mayo/Quest) ROUTINE Limbic encephalitis, new-onset seizures, psychiatric symptoms Anti-NMDAR, LGI1, CASPR2, GABA-B, AMPA
Paraneoplastic panel ROUTINE Subacute onset, smoking, weight loss Anti-Hu, Yo, CV2, amphiphysin, etc.
Cytology ROUTINE Rule out carcinomatous meningitis Negative

Note: Send paired serum for autoimmune panels - some antibodies better detected in serum vs CSF.


Subarachnoid Hemorrhage Panel

Study Priority Timing Target Finding
Cell count Tube 1 vs Tube 4 STAT Immediate Stable RBCs = true SAH; decreasing = traumatic
Xanthochromia (visual + spectrophotometry) STAT ≥6-12h from symptom onset Yellow supernatant = prior hemorrhage
Bilirubin (CSF) URGENT ≥12h from symptom onset Elevated = hemoglobin breakdown

Timing critical: Xanthochromia may not be present if LP done <6 hours from symptom onset. If initial CT and LP negative but high clinical suspicion, consider repeat LP at 12h or CTA.


Guillain-Barré Syndrome Panel

Study Priority Timing Target Finding
Protein URGENT May be normal in first week Elevated (albuminocytologic dissociation)
Cell count URGENT Diagnosis WBC <10 (if >50, reconsider diagnosis)
Glucose ROUTINE Rule out infection Normal
Cytology ROUTINE Rule out carcinomatous Negative
GQ1b antibody (serum) ROUTINE Miller Fisher variant Positive

Note: CSF protein may be normal in first 1-2 weeks of GBS. Clinical diagnosis may precede CSF abnormalities.


Carcinomatous/Lymphomatous Meningitis Panel

Study Priority Volume Needed Target Finding
Cytology URGENT 10-20 mL (large volume) Malignant cells
Flow cytometry URGENT 5-10 mL Clonal B or T cells
Protein ROUTINE Standard Often markedly elevated
Glucose ROUTINE Standard Often low
LDH ROUTINE Standard Elevated

Volume matters: Sensitivity of cytology increases with CSF volume. If first cytology negative and high suspicion, repeat LP with large volume (10-20 mL) and rapid transport (<1 hour to lab).


Normal Pressure Hydrocephalus (Therapeutic LP)

Parameter Value Notes
Volume to remove 30-50 mL Therapeutic trial
Opening pressure <24 cm H2O (by definition) Document precisely
Pre-LP gait assessment Timed 10m walk, TUG test Perform within 2h before LP
Post-LP gait assessment Repeat at 1h, 2h, 24h, 72h >20% improvement = positive
Core studies Standard panel Rule out other pathology

Positive response: ≥20% improvement in gait speed or Timed Up-and-Go (TUG) suggests shunt-responsive NPH.


Idiopathic Intracranial Hypertension (IIH) - Therapeutic LP

Parameter Value Notes
Opening pressure >25 cm H2O Diagnostic criterion
Volume to remove Until OP <20 cm H2O Therapeutic goal
CSF composition Must be normal Abnormal CSF excludes IIH
Patient position Lateral decubitus, legs extended Standardize measurement

ADVANCED MOLECULAR TESTING

Metagenomic Next-Generation Sequencing (mNGS)

Delve Detect (CSF mNGS) - UCSF-developed, commercially available

Feature Details
Pathogens detected >68,000 bacteria, viruses, fungi, parasites
Sample volume 1-2 mL CSF
Turnaround time 48 hours
Sensitivity 63% (higher than all other direct tests combined)
Specificity 99.6%
Best use case Culture-negative meningitis/encephalitis, immunocompromised, prior antibiotics
Limitations Expensive (~$2,000), may not be covered by insurance, interpretation requires expertise

When to order mNGS: - Conventional workup negative but high clinical suspicion - Immunocompromised patient with CNS infection - Patient received antibiotics before LP - Suspected rare or unusual pathogen

When NOT to order mNGS: - Undifferentiated fever without CNS signs - Routine bacterial meningitis where conventional tests adequate - As first-line test (order alongside, not instead of, conventional studies)


CSF Cell-Free DNA for CNS Tumors

MSK-IMPACT, Foundation Medicine, institutional assays

Feature Details
Indication Suspected CNS malignancy, leptomeningeal metastases
Detection rate 53% of CNS tumors with clinically documented disease
Best yield Disseminated/leptomeningeal disease, glioblastoma
Turnaround time 2-3 weeks
Sample volume 3-5 mL CSF

CSF cfDNA provides: - Molecular diagnosis when biopsy not feasible - Identification of actionable mutations (EGFR, IDH, BRAF) - Monitoring treatment response - Detection of resistance mutations

When to consider: - Suspected CNS tumor in surgically inaccessible location - Monitoring known leptomeningeal disease - Suspected tumor progression on imaging - CSF cytology negative but high suspicion


SPECIAL HANDLING REQUIREMENTS

Study Temperature Time Sensitivity Special Instructions
Cell count Room temp <1 hour Process immediately; cells lyse over time
Glucose On ice <1 hour Glycolysis continues ex vivo
Gram stain/culture Room temp <2 hours Never refrigerate
Viral PCR 4°C or frozen <24h if refrigerated Can freeze at -70°C for later testing
Cytology Room temp <1 hour Rapid transport critical for cell preservation
Oligoclonal bands 4°C Stable several days Send paired serum
mNGS Room temp <6h; 4°C <7d Per lab protocol Contact lab for frozen specimen requirements

CONTRAINDICATIONS TO LP

Absolute Contraindications

Contraindication Rationale Action
Infection at LP site Risk of introducing infection Choose alternate site or defer
Uncorrected coagulopathy (INR >1.5, platelets <50K) Bleeding risk Correct first; consider hematology consult

Relative Contraindications (Require CT First)

Clinical Feature Concern Management
Papilledema Elevated ICP, risk of herniation CT head before LP
Focal neurologic deficit Mass lesion CT head before LP
Altered consciousness Mass lesion, elevated ICP CT head before LP
Immunocompromised with new neuro symptoms Opportunistic mass lesion CT head before LP
History of CNS mass or shunt Altered CSF dynamics CT head before LP; neurosurgery consult

QUICK REFERENCE: VOLUME REQUIREMENTS

Clinical Scenario Minimum Total Volume Tube Distribution
Standard diagnostic LP 10-15 mL Tubes 1-4 (3-4 mL each)
Suspected infection (with BioFire/mNGS) 15-20 mL Extra for molecular testing
Suspected malignancy 20-30 mL Large volume for cytology
Therapeutic LP (NPH) 30-50 mL Serial collection until OP normalizes
Therapeutic LP (IIH) Until OP <20 cm H2O May require 20-40 mL

CHANGE LOG

v1.0 (January 13, 2026) - Initial creation - Added BioFire FilmArray ME Panel (14 pathogens, ~1 hour turnaround) - Added Delve Detect mNGS (>68,000 pathogens, 48h turnaround) - Added CSF cfDNA for CNS tumor diagnosis - Added NPH and IIH therapeutic LP protocols - Comprehensive diagnosis-specific panels