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 muL 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.
Author: Wanlapa RoobsoongSteven P MaherNattawan RachaphaewSamantha J BarnesKim C WilliamsonJetsumon SattabongkotJohn H Adams
Credits/Source: Malaria Journal 2014, 13:55
Published on: 2014-02-14
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News Provider: EUPB – European Press Bureau
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