Flow cytometric screening of blood samples for malaria parasites

Vianen, P. H. Van; Engen, A. Van; Thaithong, Sodsri; Keur, Maarten van der; Tanke, Hans J.; Káay, H. J. Van Der; Mons, Barend; Janse, Chris J.

doi:10.1002/cyto.990140307

Cited by 52 publications

(33 citation statements)

References 27 publications

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“…Experimental malaria-cytometry studies have found a detection threshold of 50 parasites/μL. 30 Furthermore, in a study using a Sysmex SIF flow cytometer, the authors were able to differentiate P. falciparum parasite stages based on fluorescence patterns, 31 providing evidence for the potential for improving malaria detection with commercial flow cytometers, including the XE-2100. Our findings, specially the N-OD2 models, suggest that the XE-2100 could become an accurate tool for malaria diagnosis, allowing for a timely microscopic blood evaluation of affected patients.…”

Section: Discussionmentioning

confidence: 99%

Design of Malaria Diagnostic Criteria for the Sysmex XE-2100 Hematology Analyzer

Campuzano-Zuluaga¹,

Álvarez-Sánchez²,

Escobar-Gallo³

et al. 2010

The American Society of Tropical Medicine and Hygiene

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Hygiene INTRODUCTIONEarly detection of malaria in patients not suspected of having the disease is a potential novel application for modern hematology analyzers like the Sysmex XE-2100 (Sysmex Corporation, Kobe, Kansai, Japan). A prompt and accurate diagnosis is critical to reduce adverse outcomes associated with malaria, including death. 1 In endemic regions, malaria is a prime diagnostic concern in febrile patients, and expert thickfilm evaluation is usually available.2 However, the accuracy of microscopic malaria diagnosis in less experienced centers or in non-endemic regions may be lower, and health personnel is less likely to consider the disease, leading to delayed diagnoses.1 Automated full blood counts (FBC) are usually performed in patients with febrile illnesses, providing a unique opportunity to guide a timely parasitological malaria diagnosis.Hematology analyzers evaluated for malaria detection include the Cell-Dyn (Abbott Diagnostics, Santa Clara, CA), 3 the GEN.S and LH-750 (Beckman Coulter, Miami, FL), 4 and the XE-2100 and XS-1000 i (Sysmex Corporation, Kobe, Kansai, Japan). 5,6 The Sysmex XE-2100 is an automated hematology analyzer that uses flow cytometry, direct current (DC) impedanciometry, and radiofrequency conductance (RF) to detect and measure blood corpuscular elements.7 Flow cytometry may detect malaria parasites or parasite debris such as free hemozoin or hemozoin-laden macrophages as reported for Cell-Dyn instruments.8 For the Sysmex XE-2100, two case series reported a gap between the manual and automated eosinophil counts (pseudoeosinophilia) and abnormal granulocyte-coded events in the DIFF scatterplot used for white blood cell (WBC) separation based on fluorescence versus side-scatter signals in samples from Plasmodium vivaxinfected patients.9, 10 Later, Huh and others 5 found that the two previously reported abnormalities, compared with thick film, had a sensitivity of 69.4% and a specificity of 100% for P. vivax diagnosis. These findings suggest the potential clinical utility of malaria detection by the XE-2100. Ideally, to improve the clinical impact of this detection method, it should be an automated process relying on the analyzer's quantitative data and expressed as a "malaria alarm" that could motivate the microscopic search for malaria.The primary aim of this study was to develop, validate, and test the accuracy of one observer-dependent (OD) and two non-observer-dependent (N-OD) diagnostic prediction models for P. vivax and P. falciparum malaria. The approach for developing the OD models was similar to previous studies using the XE-2100 for malaria diagnosis 5 with the inclusion of other clinical and scatterplot variables not previously investigated. The N-OD models were developed to obtain a framework for an automated malaria-detection algorithm for the XE-2100. Secondary aims were to determine the occurrence of pseudoeosinophilia and to explore a possible cause for the XE-2100 FBC malaria-related abnormalities. MATERIALS AND METHODSStudy population. Blood samples were col...

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Section: Discussionmentioning

confidence: 99%

Design of Malaria Diagnostic Criteria for the Sysmex XE-2100 Hematology Analyzer

Campuzano-Zuluaga¹,

Álvarez-Sánchez²,

Escobar-Gallo³

et al. 2010

The American Society of Tropical Medicine and Hygiene

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“…Flow cytometry has been successfully used to analyze parasite development by determining the amount of DNA synthesized using DNA-specific dyes (37). By simultaneously staining for erythrocyte surface CD59 and parasite DNA, we are able to assess intraerythrocytic infection by the malaria parasite.…”

Section: Movement Of Erythrocyte Raft Proteins and Lipids Into A Malamentioning

confidence: 99%

The Role of Cholesterol and Glycosylphosphatidylinositol-anchored Proteins of Erythrocyte Rafts in Regulating Raft Protein Content and Malarial Infection

Samuel

Mohandas²,

Harrison

et al. 2001

Journal of Biological Chemistry

173

152

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Human erythrocytes are terminally differentiated, nonendocytic cells that lack all intracellular organelles. Here we show that their plasma membranes contain detergentresistant membrane rafts that constitute a small fraction (4%) of the total membrane protein, with a complex mixture of proteins that differentially associate with rafts. Depletion of raft-cholesterol abrogates association of all proteins with no significant effect on cholesterol:protein ratios in the rest of the membrane, lipid asymmetry, deformability, or transport properties of the bilayer, indicating that cholesterol is critical for protein assembly into rafts and suggesting that rafts have little influence on several erythrocyte functions. Erythrocytes from patients with paroxysmal nocturnal hemoglobinuria, which lack glycosylphosphatidylinositol-anchored proteins, show significant elevation in raft-cholesterol but no increase in raft protein association, suggesting that raft assembly does not require glycosylphosphatidylinositol-anchored proteins, raft proteins do not bind directly to cholesterol, and only threshold levels of raft-cholesterol are critical for protein recruitment. Loss of glycosylphosphatidylinositol-anchored proteins had no effect on erythrocytic infection by malarial parasite or movement of raft markers into the parasite's vacuole. However, infection is blocked following raft-cholesterol disruption, suggesting that erythrocyte rafts can be functionally exploited and providing the first evidence for the involvement of host rafts in an apicomplexan infection.

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“…30 Other diagnostic methods include automated detection of malaria pigment 38 and flow cytometry. 39 Serologic tests have no role in the diagnosis of acute malaria.…”

Section: Diagnosismentioning

confidence: 99%

Malaria

Suh¹

2004

Canadian Medical Association Journal

103

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Malaria is a parasitic infection of global importance. Although relatively uncommon in developed countries, where the disease occurs mainly in travellers who have returned from endemic regions, it remains one of the most prevalent infections of humans worldwide. In endemic regions, malaria is a significant cause of morbidity and mortality and creates enormous social and economic burdens. Current efforts to control malaria focus on reducing attributable morbidity and mortality. Targeted chemoprophylaxis and use of insecticide-treated bed nets have been successful in some endemic areas. For travellers to malaria-endemic regions, personal protective measures and appropriate chemoprophylaxis can significantly reduce the risk of infection. Prompt evaluation of the febrile traveller, a high degree of suspicion of malaria, rapid and accurate diagnosis, and appropriate antimalarial therapy are essential in order to optimize clinical outcomes of infected patients. Additional approaches to malaria control, including genetic manipulation of mosquitoes and malaria vaccines, are areas of ongoing research

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Flow cytometric screening of blood samples for malaria parasites

Cited by 52 publications

References 27 publications

Design of Malaria Diagnostic Criteria for the Sysmex XE-2100 Hematology Analyzer

Design of Malaria Diagnostic Criteria for the Sysmex XE-2100 Hematology Analyzer

The Role of Cholesterol and Glycosylphosphatidylinositol-anchored Proteins of Erythrocyte Rafts in Regulating Raft Protein Content and Malarial Infection

Malaria

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