Total IgA and norovirus-specific salivary IgA titers were determined using GII.4 Den Haag VLPs for baseline I, GII.4 New Orleans VLPs for baseline II, or the outbreak-specific VLP [24 (link)]. Briefly, 96-well plates were coated with 1 μg/mL anti-human IgA or 1 μg/mL VLPs. Two-fold serial dilutions of saliva and purified human IgA (Sigma) as a standard were tested in duplicate. Bound IgA was detected using goat anti-human IgA-horseradish peroxidase (HRP) conjugate followed by color development with 3,3′,5,5′-tetramethylbenzidine (TMB) substrate (Thermo Scientific).
Protocol detail
Quantifying Salivary IgA Titers
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Corresponding Organization : Centers for Disease Control and Prevention
Other organizations : Oak Ridge Associated Universities, Oregon Health Authority
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Variable analysis
independent variables
- GII.4 Den Haag VLPs
- GII.4 New Orleans VLPs
- Outbreak-specific VLP
- Total IgA titers
- Norovirus-specific salivary IgA titers
- Concentration of anti-human IgA (1 μg/mL)
- Concentration of VLPs (1 μg/mL)
- Two-fold serial dilutions of saliva
- Purified human IgA (Sigma) as a standard
- Goat anti-human IgA-horseradish peroxidase (HRP) conjugate
- 3,3′,5,5′-tetramethylbenzidine (TMB) substrate (Thermo Scientific)
- Positive control: Purified human IgA (Sigma) as a standard
- Negative control: Not explicitly mentioned
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