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Preparation of Solid Tissue for Aspergillus Galactomannan

时间:2006-07-24 14:26来源:Scienceboard.net 作者:admin
Thursday, November 13, 2003

Description
Platelia Aspergillus (Biorad) is a 1-stage immunoenzymatic sandwich microplate kit, for the detection of antigen galactomannan

Procedure
In a laminar flow safety cabinet (class II), fine cut the sample with a sterile scalpel in a Petri dish. It is important to cut the samples in all directions: length, width and diagonal to break the tissue fibres and to optimise the extraction of the antigen
Transfer the small pieces in the sterile glass homogeniser (1 - 5ml, depending on volume of specimen) and grind it to get an homogenate
Add extra pure water, of equal volume to the sample. This helps grinding and to get a liquid homogenate
Aliquot in one or several sterile eppendorf/microtubes of 1.5 ml capacity, 1 ml per aliquot.
Vortex the homogenate for 30 seconds
Centrifuge at 1000g for 5 minutes
Remove the supernatant in a tube with a sterile pipette tip
Analyse the supernatant with the same technique used for serum following the kits directions

Notes
Minimise contact of specimen with dust/air, due to the sensitivity of the Platelia test (1ng of galactomannan per ml). Any possibility of contamination during the various steps of the procedure must be prevented in order to avoid false positive results.



Recipes
Equipment
safety cabinet
sterile Petri dish
sterile scalpel
sterile glass homogeniser
extra pure water (as recommended for use in assay)
1.5ml sterile eppendorfs/microtubes
vortex
centrifuge
sterile pipette tips

Ideally pyrogen free equipment should be used, although standard sterile equipment should be sufficient (following manufacturers advice).

Health and Safety
perform procedure in a class II laminar flow safety cabinet
wear disposable gloves


Supplies


Tips

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