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RT-QuIC Nasal Brushing

A Test for Creutzfeldt–Jakob Disease Using Nasal Brushings

Year of Publication: 2014

Authors: Christina D. Orrú, Matilde Bongianni, Giovanni Tonoli, ..., Gianluigi Zanusso

Journal: New England Journal of Medicine

Citation: N Engl J Med 2014;371:519-29

Link: https://www.nejm.org/doi/full/10.1056/NEJMoa1315200


Clinical Question

Can RT-QuIC analysis of olfactory epithelium samples obtained via nasal brushings accurately diagnose Creutzfeldt-Jakob disease in living patients?

Bottom Line

RT-QuIC testing of nasal brushings demonstrated 97% sensitivity and 100% specificity for diagnosing CJD, outperforming CSF-based RT-QuIC (77% sensitivity) in the same patients. This minimally invasive approach detects substantial prion seeding activity in the olfactory epithelium and offers a promising method for definitive antemortem diagnosis of CJD.

Major Points

  • RT-QuIC of nasal brushings positive in 30/31 CJD patients (15/15 definite sCJD, 13/14 probable sCJD, 2/2 inherited E200K CJD)
  • All 43 non-CJD controls were RT-QuIC negative on nasal brushings
  • Olfactory mucosa samples gave significantly stronger and faster RT-QuIC responses than CSF (P<0.001)
  • Individual brushings contained approximately 10⁵ to 10⁷ prion seeds, several logs₁₀ greater concentration than in CSF
  • Nasal brushing is a safe, minimally invasive procedure requiring only local vasoconstrictor and fiberoptic rhinoscope
  • Method detected all CJD molecular subtypes (MM, MV, VV genotypes; types 1 and 2 PrPᶜᴶᴰ)
  • One false negative case (Patient 29) had typical tau and 14-3-3 levels but negative RT-QuIC on both OM and CSF
  • The presence of substantial prion seeds in the nasal vault raises biosafety considerations for medical instruments contacting this area

Design

Study Type: Prospective diagnostic accuracy study

Randomization:

Blinding: Blinded analysis; RT-QuIC testing performed on blinded sample sets

Enrollment Period: Not specified

Follow-up Duration: Not applicable (diagnostic study)

Centers: 2

Countries: Italy, United States

Sample Size: 74

Analysis: Sensitivity and specificity calculated with 95% confidence intervals using Wilson procedure; statistical comparison of ThT fluorescence responses using unpaired t-tests


Inclusion Criteria

  • CJD patients: Rapidly progressive dementia referred due to possible or probable CJD
  • Classified according to updated clinical diagnostic criteria (Puoti et al. 2012)
  • Controls with other neurological disorders: Probable Alzheimer's disease, probable Parkinson's disease, progressive supranuclear palsy, paraneoplastic limbic encephalitis
  • Controls without neurological disorders: Persons referred to ENT clinic for other purposes

Exclusion Criteria

  • Not explicitly stated

Arms

FieldCJD PatientsControl
InterventionOlfactory mucosa obtained via nasal brushing; CSF obtained via lumbar puncture; samples analyzed by RT-QuIC assayOlfactory mucosa obtained via nasal brushing; CSF obtained via lumbar puncture (subset); samples analyzed by RT-QuIC assay
DurationSingle time point diagnostic testingSingle time point diagnostic testing

Outcomes

OutcomeTypeControlInterventionHR / OR / RRP-value
Diagnostic accuracy of RT-QuIC assay on olfactory mucosa samples for detecting CJDPrimary
RT-QuIC sensitivity on CSF samplesSecondary
RT-QuIC sensitivity in definite sporadic CJD (olfactory mucosa)Secondary
RT-QuIC sensitivity in probable sporadic CJD (olfactory mucosa)Secondary
Mean ThT fluorescence comparison OM vs CSFSecondary<0.001
Prion seed concentration in olfactory mucosaSecondary
Prion seed concentration in CSFSecondary
Prion seed concentration in brain tissueSecondary
14-3-3 assay sensitivitySecondary
CSF tau >2400 pg/mL sensitivitySecondary
Nasal brushing procedureAdverse

Subgroup Analysis

RT-QuIC detected all CJD molecular subtypes: MM genotype (14/14), MV genotype (8/9), VV genotype (2/2), and both type 1 and type 2 PrPᶜᴶᴰ. The single false negative (Patient 29, MV genotype) was RT-QuIC negative on both OM and CSF despite having clinical features and biomarkers typical of sporadic CJD.


Criticisms

  • Relatively small sample size (31 CJD patients, 43 controls); larger validation studies needed
  • Single-center sampling (Italy) for CJD patients limits generalizability
  • One false negative case suggests method may not detect all CJD cases
  • Time between symptom onset and OM brushing varied widely (0.5-28 months); effect of disease stage on test performance unclear
  • Control population had significantly different age distribution than CJD patients
  • Biosafety implications of prion-contaminated medical instruments not fully addressed
  • Study did not compare performance to brain biopsy (true gold standard in living patients)
  • Only sporadic and E200K inherited CJD tested; performance in other inherited forms unknown
  • Requires specialized laboratory expertise and equipment for RT-QuIC assay

Funding

Intramural Research Program of the National Institute of Allergy and Infectious Diseases (NIAID); Fondazione Cariverona; Italian Ministry of Health (RF2009-1474758); Creutzfeldt-Jakob Disease Foundation; Programma Master and Back–Percorsi di rientro fellowship; NIAID Gift Fund donations

Based on: RT-QuIC Nasal Brushing (New England Journal of Medicine, 2014)

Authors: Christina D. Orrú, Matilde Bongianni, Giovanni Tonoli, ..., Gianluigi Zanusso

Citation: N Engl J Med 2014;371:519-29

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