Re: anti-malarial antibodies

From: Gabriel Alespeiti (gabriel_alespeiti@nybc.org)
Date: Fri Jul 26 2002 - 19:40:20 EST


Dear colleagues,

I have read the replies on anti-malarial antibodies and I hope to get
some help from the experts in this field. A researcher in the
Institute is conducting parasitaemia studies of Plasmodium falciparum
infected erythrocytes using Giemsa stained microscopy, a tedious,
time consuming and technically imprecise method. We want to replace
the microscopic counts with a flow cytometry method. Peripheral blood
will be collected in Brazil and will need to be fixed before
shipping. A FACScalibur is available for this work. We would like to
analyze 25 samples a day, the expected parasitemia level is between
0.05% and 8%.
I tried formaldehyde fixation followed by propidium iodide staining
and even though there is a direct correlation between cytometric and
microscopic determination,  the manual counts are significantly
higher than the flow counts.
What I'm looking for is a simple, proven protocol for daily use that
gives directly the % of parasitaemia by flow. In our case,
determination of erythrocyte or parasite antigens; even though
useful, is not necessary. What is important for us is to be able to
detect low infection levels.
Does anybody know, what is the lowest level of infection that can be
detected by flow?
Please post the reply on this site, so other people can benefit from
it. Thank you very much.





--
**************************************
Gabriel E. Alespeiti
Supervisor, Flow Cytometry
Lindsley F. Kimball Research Institute
New York Blood Center
310 E.67th St. Room 2-16E
New York, NY 10021

Phone: (212) 570-3346
Fax: (212) 570-3307
E-mail:gabriel_alespeiti@nybc.org
**************************************



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