Sample extraction for quantification of mold antigen

Instrument: Precellys® 24 Dual with Cryolys cooling device (+4◦C).

Sample: Teflon filters with airborne dust

Protocol:

Precellys kits: SK38_2ml (03961-1-006), SK38_7ml (03961-1-303)
Samples: Parallel sampled Teflon filters with airborne dust from composting plants.
Phosphate buffered saline with 0.05% Tween 20.
Run the Precellys 24 Dual at 6000 rpm, 3x20 sec, 30 sec break with cooling vs mixing only.
Enzyme immunoassays: Aspergillus fumigatus and Penicillium chrysogenum based on polyclonal rabbit antibodies.

Results:

For extraction of airborne dust parallel sampled on 10 Teflon filters in composting plants, the SK38 - 6000 rpm homogenization procedures (mixed only, Precellys homogenization, with filter removed or not) were applied. With Precellys SK38 homogenization, A. fumigatus and P. chrysogenum antigen yields were higher than by mixing only irrespective of the mode of filter removal. Higher antigen amounts were obtained for both volumes of homogenization: 2mL (Fig. 1) or 7mL kit, with 1mL or 3mL of buffer, respectively.

The antigen amounts of samples with fungal material were increased after extraction and homogenization with the combo Precellys 24 Dual & Cryolys and SK38 kit.

In 5 composting plants, airborne dust was sampled on Teflon filters with a Parallel sampler. The Aspergillus fumigatus (a) or Penicillium chrysogenum (b) antigen yields after extraction in 2 ml Precellys tubes were measured by enzyme immunoassays (mean of 2 filters per procedure).