Abstract:
BACKGROUND:Plasmodium vivax preferentially infects Duffy-positive reticulocytes and infections typically have few parasite-infected cells in the peripheral circulation. These features complicate detection and quantification by flow cytometry (FC) using standard nucleic acid-based staining methods. A simple antibody-based FC method was developed for rapid parasite detection along with simultaneous detection of other parasite and erythrocyte markers. METHODS:Clinical samples were collected from patients diagnosed with P. vivax at a district Malaria Clinic in Kanchanaburi, Thailand. One μL of infected blood was washed, fixed, stained with a Plasmodium pan-specific anti-PfBiP antibody conjugated with Alexa Fluor 660, and analysed by FC. Additional primary conjugated antibodies for stage-specific markers of P. vivax for late trophozoite-early schizonts (MSP1-Alexa Fluor 660), late-stage schizonts (DBP-Alexa Fluor 555), and sexual stages (Pvs16) were used to differentiate intra-erythrocytic developmental stages. RESULTS:The percentages of P. vivax-infected cells determined by the FC method and manually by microscopic examination of Giemsa-stained thick blood smears were positively correlated by Spearman's rank correlation coefficient (R2=0.93843) from 0.001 to 1.00% P. vivax-infected reticulocytes. CONCLUSIONS:The FC-based method is a simple, robust, and efficient method for detecting P. vivax-infected reticulocytes.
journal_name
Malar Jjournal_title
Malaria journalauthors
Roobsoong W,Maher SP,Rachaphaew N,Barnes SJ,Williamson KC,Sattabongkot J,Adams JHdoi
10.1186/1475-2875-13-55subject
Has Abstractpub_date
2014-02-14 00:00:00pages
55issn
1475-2875pii
1475-2875-13-55journal_volume
13pub_type
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