A rapid sensitive, flow cytometry-based method for the detection of Plasmodium vivax-infected blood cells.

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 J

journal_title

Malaria journal

authors

Roobsoong W,Maher SP,Rachaphaew N,Barnes SJ,Williamson KC,Sattabongkot J,Adams JH

doi

10.1186/1475-2875-13-55

subject

Has Abstract

pub_date

2014-02-14 00:00:00

pages

55

issn

1475-2875

pii

1475-2875-13-55

journal_volume

13

pub_type

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