✓Prepare pneumococcal inoculum for controlled human nasal colonization
✓Perform nasal inoculation and monitor volunteer responses
✓Execute nasal wash sampling and culture-based detection
✓Interpret bacterial carriage outcomes from blood agar plates
Protocol
Experimental human pneumococcal carriage offers a natural model of carriage and a potential model for use in vaccine development. This technique is valuable yet complex and involves clinical risk by introducing a pathogen into a human. We have developed a detailed protocol.
Difficulty
advanced
Total time
~3–7 days per volunteer (including inoculation, serial sampling, and culture analysis)
Model organism
Human volunteer
Biosafety
BSL-2
Steps
1
Prepare pneumococcal inoculum and inoculate volunteer
Culture and prepare pneumococcal suspension to specific viable count, then administer controlled nasal inoculation to consenting volunteer. Monitor volunteer tolerance and early clinical responses.
▶ 01:41
2
Collect nasal wash samples from inoculated volunteer
Perform serial nasal wash procedure post-inoculation to recover colonizing bacteria from nasal mucosa. Timing and repetition critical for carriage kinetics.
▶ 03:41
3
Process nasal wash samples for bacterial culture
Plate nasal wash samples on pneumococcal-selective media (blood agar) and incubate under appropriate conditions to isolate and enumerate carriage bacteria.
▶ 04:35
4
Analyze carriage outcomes and clinical symptoms
Quantify bacterial colony counts from culture plates and correlate with reported volunteer symptoms to assess carriage establishment and clinical burden.
▶ 06:11
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