Improved Models of Mini Anion Exchange Centrifugation Technique (mAECT) and Modified Single Centrifugation (MSC) for Sleeping Sickness Diagnosis and Staging

Büscher, Philippe; Ngoyi, Dieudonné Mumba; Kaboré, Jacques; Lejon, Veerle; Robays, Jo; Jamonneau, Vincent; Bebronne, Nicolas; Veken, Wim Van der; Biéler, Sylvain

doi:10.1371/journal.pntd.0000471

Cited by 106 publications

(95 citation statements)

References 9 publications

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“…Cases were defined as CATT positive individuals in whose body fluids (blood, lymph, or CSF) trypanosomes had been respectively demonstrated by the haematocrit centrifugation technique (HCT) (Woo, 1971), wet films or modified single centrifugation (MSC) (Büscher et al, 2009), respectively. These 3 tests represented the composite reference standard.…”

Section: Case and Control Definitionsmentioning

confidence: 99%

Comparison of nucleic acid sequence-based amplification and loop-mediated isothermal amplification for diagnosis of human African trypanosomiasis

Mugasa

Katiti

Boobo

et al. 2014

Diagnostic Microbiology and Infectious Disease

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Section: Case and Control Definitionsmentioning

confidence: 99%

Comparison of nucleic acid sequence-based amplification and loop-mediated isothermal amplification for diagnosis of human African trypanosomiasis

Mugasa

Katiti

Boobo

et al. 2014

Diagnostic Microbiology and Infectious Disease

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“…At first-peak parasitemia, when the blood contained 10 8 /ml long, slender bloodstream form trypomastigotes, the rat was anesthetized, and the blood was collected by cardiac puncture with a heparinized syringe. Trypanosomes were separated from the blood by the mini-anion exchange centrifugation technique (mAECT) (21) and centrifuged at 1,000 ϫ g for 15 min. The parasites were then resuspended at a concentration of 10 5 cells/ml in HMI-9 medium supplemented with 15% fetal calf serum and used to initiate in vitro culture according to Van Reet et al (22).…”

Section: Ethicsmentioning

confidence: 99%

Killing of Trypanozoon Parasites by the Equine Cathelicidin eCATH1

Cauchard

Reet

Büscher

et al. 2016

Antimicrob Agents Chemother

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h Trypanozoon parasites infect both humans, causing sleeping sickness, and animals, causing nagana, surra, and dourine. Control of nagana and surra depends to a great extent on chemotherapy. However, drug resistance to several of the front-line drugs is rising. Furthermore, there is no official treatment for dourine. Therefore, there is an urgent need to develop antiparasitic agents with novel modes of action. Host defense peptides have recently gained attention as promising candidates. We have previously reported that one such peptide, the equine antimicrobial peptide eCATH1, is highly active against equine Gram-positive and Gram-negative bacteria, without cytotoxicity against mammalian cells at bacteriolytic concentrations. In the present study, we show that eCATH1 exhibits an in vitro 50% inhibitory concentration (IC 50 ) of 9.5 M against Trypanosoma brucei brucei, Trypanosoma evansi, and Trypanosoma equiperdum. Its trypanocidal mechanism involves plasma membrane permeabilization and mitochondrial alteration based on the following data: (i) eCATH1 induces the rapid influx of the vital dye SYTOX Green; (ii) it rapidly disrupts mitochondrial membrane potential, as revealed by immunofluorescence microscopy using the fluorescent dye rhodamine 123; (iii) it severely damages the membrane and intracellular structures of the parasites as early as 15 min after exposure at 9.5 M and 5 min after exposure at higher concentrations (19 M), as evidenced by scanning and transmission electron microscopy. We also demonstrate that administration of eCATH1 at a dose of 10 mg/kg to T. equiperdum-infected mice delays mortality. Taken together, our findings suggest that eCATH1 is an interesting template for the development of novel therapeutic agents in the treatment of trypanosome infections.

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“…However, its limited specificity results in a substantial number of nonconfirmed seropositive individuals and in some disease foci, the sensitivity of the CATT is rather low [11]. The mAECT technique is the most sensitive trypanosome detection format with a detection limit of <50 parasites per ml blood [12,13]. Despite its high sensitivity and specificity, large-scale use of mAECT is hampered by its high cost and irregular production.…”

Section: Reviewmentioning

confidence: 99%

Recent progress in molecular diagnosis of sleeping sickness

Deborggraeve¹,

Büscher²

2012

Expert Review of Molecular Diagnostics

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This article will review the most recent progress in the molecular diagnosis of sleeping sickness and its potential role in patient management and disease control. While PCR remains restricted to research and reference laboratories, promising alternative molecular platforms have emerged over the last few years. Several loop-mediated isothermal amplification assays have been developed for detection and identification of the parasite with reported high analytical sensitivity and specificity. Simplified loop-mediated isothermal amplification formats have been designed and are undergoing evaluation studies in the field. Accurate diagnosis based on specific detection of the parasite's ribosomal RNA has been made possible by the isothermal nucleic acid sequence-based amplification and by direct hybridization with fluorescent detection probes. In addition to the technological progress, the authors also discuss the diagnostic performance of molecular tests in the most recent clinical evaluation studies and briefly present some viewpoints for the near future.

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Improved Models of Mini Anion Exchange Centrifugation Technique (mAECT) and Modified Single Centrifugation (MSC) for Sleeping Sickness Diagnosis and Staging

Cited by 106 publications

References 9 publications

Comparison of nucleic acid sequence-based amplification and loop-mediated isothermal amplification for diagnosis of human African trypanosomiasis

Comparison of nucleic acid sequence-based amplification and loop-mediated isothermal amplification for diagnosis of human African trypanosomiasis

Killing of Trypanozoon Parasites by the Equine Cathelicidin eCATH1

Recent progress in molecular diagnosis of sleeping sickness

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