1. A rapid sensitive, flow cytometry-based method for the detection of Plasmodium vivax-infected blood cells.
- Author
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Roobsoong W, Maher SP, Rachaphaew N, Barnes SJ, Williamson KC, Sattabongkot J, and Adams JH
- Subjects
- Antibodies, Protozoan, Antigens, Protozoan analysis, Fluorescent Dyes analysis, Humans, Staining and Labeling, Thailand, Blood Cells parasitology, Flow Cytometry methods, Malaria, Vivax diagnosis, Plasmodium vivax isolation & purification
- 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.
- Published
- 2014
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