Whilst we have also done agglutination assays on the XL, you
can test antisera pre and post vaccination by labeling the
bacteria directly with the serum followed by FITC labeled
anti human Ig (on which there is some literature). This is
how we test the specificity of our antibodies. Your best
bet on the XL is to trigger on side scatter as an auxiliary
signal. The most convenient sample preparation is by
staining in 96-well filter plates were your bugs are
retained on the membrane and can be resuspendet for
measurement.
Regards
Gerhard Nebe-v.Caron
Unilever Research, Colworth,
Sharnbrook, Bedfordshire
GB - MK44 1LQ
Tel: +44(0)1234-222066
FAX: +44(0)1234-222344
gerhard.nebe-von-caron@unilever.com
______________________________ Reply Separator _________________________________
Subject: qualitative bacterial agglutination by flow
Author: JHUGHES@ICH.UCT.AC.ZA at INTERNET
Date: 24/11/97 23:21
Hi all Flowers
Our laboratory has been trying to perform a qualitative agglutination
assay for Bordetella Pertussis in pre and post vaccinated sera using
a visual end point. The results are not satisfactorily reproducible
from one reader to another. The end point is very subtle; a fine
microagglutination that is recommended to be read under oblique
light with a magnifying glass.
Have any of you any experience with bacterial agglutination by flow.
We have a Coulter XL; I have no prior experience with 'bugs' and
would need some guidance as to recommended settings etc.
I am VERY keen to eliminate the human error apparently inherent in
this assay...am I asking the impossible!!!??
Jane Hughes
Chief, Institute of Child Health Lab,
Red Cross Children's Hospital
Rondebosch, Cape Town 7700.
South Africa
Tel 021 658 5315
Fax 021 689 1287
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