Document Detail


Re-evaluating acridine orange for rapid flow cytometric enumeration of parasitemia in malaria-infected rodents.
MedLine Citation:
PMID:  17421026     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Methods facilitating research in malaria are of pivotal relevance. Flow cytometry offers the possibility of rapid enumeration of parasitemia. It relies on staining the parasite DNA to distinguish between infected and non-infected red blood cell (RBC) populations. Unfortunately, in rodents abundant reticulocyte RNA interferes with the application of the method. This results in time-consuming sample preparation protocols that offer no clear advantage over microscopic counting. We re-evaluated the use of the DNA/RNA discriminating vital fluorochrome acridine orange (AO) for rapid flow cytometric enumeration of parasitemia in rodents. Whole blood from rodents infected with Plasmodium berghei and Plasmodium yoelii was stained with AO and analyzed by flow cytometer. A newly developed two-channel (FL1/FL3) detection method was compared with conventional one-channel (FL1) detection and microscopic counting. The new AO two-channel detection method clearly discriminated between infected and non-infected RBC populations. It showed to be linear above parasitemias of 0.3%. Sample processing time amounted to approximately 5 min. It is shown that AO can be used for rapid, precise, and accurate enumeration of parasitemia in rodents. Due to its ease of handling the method might find widespread application in malaria research.
Authors:
Sebastian Chakrit Bhakdi; Panudda Sratongno; Pattamawan Chimma; Thanaporn Rungruang; Aporn Chuncharunee; Hartmut P H Neumann; Prida Malasit; Kovit Pattanapanyasat
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cytometry. Part A : the journal of the International Society for Analytical Cytology     Volume:  71     ISSN:  1552-4922     ISO Abbreviation:  Cytometry A     Publication Date:  2007 Sep 
Date Detail:
Created Date:  2007-08-27     Completed Date:  2007-11-28     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101235694     Medline TA:  Cytometry A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  662-7     Citation Subset:  IM    
Copyright Information:
Copyright (c) 2007 International Society for Analytical Cytology.
Affiliation:
Division of Instruments for Research, Office for Research and Development, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.
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MeSH Terms
Descriptor/Qualifier:
Acridine Orange*
Animals
Cricetinae
Erythrocytes / parasitology*
Flow Cytometry / methods*
Fluorescent Dyes*
Malaria / diagnosis,  parasitology*
Mesocricetus
Mice
Mice, Inbred ICR
Parasitemia / parasitology
Plasmodium berghei / isolation & purification*,  parasitology
Plasmodium yoelii / isolation & purification*,  parasitology
Staining and Labeling
Chemical
Reg. No./Substance:
0/Fluorescent Dyes; 65-61-2/Acridine Orange
Comments/Corrections
Comment In:
Cytometry A. 2007 Sep;71(9):643-5   [PMID:  17712779 ]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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