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

Malaria Journal

Methodology

Kim C Williamson¹  Samantha J Barnes²  Steven P Maher²  John H Adams²  Wanlapa Roobsoong³  Nattawan Rachaphaew⁴  Jetsumon Sattabongkot⁴ 

[1] Department of Biology, Loyola University, 60660, Chicago, IL, USA;Department of Global Health, College of Public Health, University of South Florida, Tampa, FL, USA;Department of Global Health, College of Public Health, University of South Florida, Tampa, FL, USA;Mahidol Vivax Research Unit, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand;Mahidol Vivax Research Unit, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand;

关键词: Malaria;    Vivax malaria;    Plasmodium vivax;    Flow cytometry;    Diagnosis;   

DOI  :  10.1186/1475-2875-13-55

 received in 2013-10-02, accepted in 2014-01-22,  发布年份 2014

来源: Springer

【 摘 要 】

BackgroundPlasmodium 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.MethodsClinical 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.ResultsThe 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.ConclusionsThe FC-based method is a simple, robust, and efficient method for detecting P. vivax-infected reticulocytes.

【 授权许可】

Unknown   
© Roobsoong et al.; licensee BioMed Central Ltd. 2014. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.