Preparation of Solid Tissue for Aspergillus Galactomannan

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

(责任编辑：泉水)

搜索

热门标签:

Preparation of Solid Tissue for Aspergillus Galactomannan