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Journal of Clinical Microbiology, September 2005, p. 4789-4795, Vol. 43, No. 9
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.9.4789-4795.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Dot Enzyme-Linked Immunosorbent Assay for More Reliable Staging of Patients with Human African Trypanosomiasis

Equipe Accueil 3174 "Neuroparasitologie et neuroépidémiologie tropicale," Institut d'épidémiologie neurologique et de neurologie tropicale, Faculty of Medicine, Limoges, France,¹ Department of Parasitology, Centre International de Recherche Médicale de Franceville (CIRMF), Franceville, Gabon,² Department of Neurology, Hôpital de l'Amitié, Bangui, Central African Republic,³ Programme National de Lutte contre la Trypanosomose Humaine Africaine (PNLTHA), Ministère de la Santé Publique, Bangui, Central African Republic,⁴ Instituto de Combate e Controlo das Tripanossomiases (ICCT), Ministério da Saúde, Luanda, Angola,⁵ Equipe Accueil 3842 "Homéostasie cellulaire et pathologies," Faculty of Medicine, Limoges, France⁶

Received 11 April 2005/ Returned for modification 13 May 2005/ Accepted 7 June 2005

Human African trypanosomiasis (HAT) or sleeping sickness is a disease characterized by a hemolymphatic stage 1 followed by a meningoencephalitic stage 2 which is fatal without specific treatment. Furthermore, due to the toxicity of drugs used to treat stage 2 (mainly melarsoprol) accurate staging is required. Actual criteria employed during field surveys are not sensitive enough for precise staging. Antineurofilament (anti-NF) and antigalactocerebrosides (anti-GalC) antibodies have been identified in cerebrospinal fluid (CSF) as potential markers of central nervous system (CNS) involvement. We describe a dot enzyme-linked immunosorbent assay (dot-ELISA) to detect anti-GalC and anti-NF antibodies and its value in staging. NF- and GalC-dotted nitrocellulose strips were first developed in our laboratory. They were then evaluated in Angola and Central African Republic on 140 CSF samples. Compared to our staging criteria (i.e., CSF cell count 20 cells/µl, CSF immunoglobulin M concentration 100 mg/liter, and/or the presence of trypanosomes in the CSF), combined detection of both CSF anti-NF and CSF anti-GalC by dot-ELISA showed 83.2% sensitivity and 100.0% specificity. Dot-ELISA could be a useful test to diagnose CNS involvement in HAT in the less-equipped laboratories or in the field situation and to improve patient treatment.

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