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

Abstract

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/l, 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.

Controlling sleeping sickness – a review

“…There are also several promising developments in disease biomarkers (Papadopoulos et al 2004 ;Agranoff et al 2005). Blood or CSF markers for staging are in the early stages of development (Lejon et al 2002 ;Courtioux et al 2005) as are non-invasive diagnostic tools (Lejon et al 2006).…”

Human African Trypanosomiasis

“…106 Antibodies against neurofi laments, galactocerebrosides, neurofi laments, and glial fi brillary acidic proteins are also promising CSF markers of second-stage disease. [107][108][109] The diagnostic approach for T b rhodesiense disease diff ers from that for T b gambiense in several ways. First, there is no serological screening test for T b rhodesiense.…”

Biosynthesis and Immunogenicity of Glucosylceramide in Cryptococcus neoformans and Other Human Pathogens

“…Patients with central nervous system involvement during trypanosomiasis have been shown to have antibodies to galactocerebrosides in the cerebrospinal fluid that react with bovine galactocerebrosides (19). As in C. jejuni infection, in trypanosomiasis caused by Trypanosoma brucei, it has been suggested that anti-galactocerebroside antibodies cross-react with their mammalian counterparts (19). Furthermore, sera from patients infected with T. cruzi react with GSLs as well, although the specificity of those antibodies seems quite diverse (9).…”

“…Many Trypanosoma species contain neutral GSLs (97). Patients with central nervous system involvement during trypanosomiasis have been shown to have antibodies to galactocerebrosides in the cerebrospinal fluid that react with bovine galactocerebrosides (19). As in C. jejuni infection, in trypanosomiasis caused by Trypanosoma brucei, it has been suggested that anti-galactocerebroside antibodies cross-react with their mammalian counterparts (19).…”