Abstract
BackgroundDiagnosis of Trypanosoma cruzi infection by direct pathogen detection is complicated by the low parasite burden in subjects persistently infected with this agent of human Chagas disease. Determination of infection status by serological analysis has also been faulty, largely due to the lack of well-characterized parasite reagents for the detection of anti-parasite antibodies.MethodsIn this study, we screened more than 400 recombinant proteins of T. cruzi, including randomly selected and those known to be highly expressed in the parasite stages present in mammalian hosts, for the ability to detect anti-parasite antibodies in the sera of subjects with confirmed or suspected T. cruzi infection.FindingsA set of 16 protein groups were identified and incorporated into a multiplex bead array format which detected 100% of >100 confirmed positive sera and also documented consistent, strong and broad responses in samples undetected or discordant using conventional serologic tests. Each serum had a distinct but highly stable reaction pattern. This diagnostic panel was also useful for monitoring drug treatment efficacy in chronic Chagas disease.ConclusionsThese results substantially extend the variety and quality of diagnostic targets for Chagas disease and offer a useful tool for determining treatment success or failure.
Highlights
Chagas disease, a consequence of infection by the protozoan parasite Trypanosoma cruzi, affects up to 20 million individuals primarily in the Americas where the insect vectors are present and where zoonotic transmission cycles guarantee a steady source of parasites
As part of a vaccine discovery effort, nearly 1500 genes from T. cruzi have been cloned into Gateway entry vector plasmids that allow them to be moved into a range of other plasmids
Genes were selected for cloning using a variety of criteria, initially including known expression in T. cruzi lifecycle stages that are present throughout infection in mammals, high likelihood of being surface expressed or secreted and expected presence in the genome at low copy number
Summary
A consequence of infection by the protozoan parasite Trypanosoma cruzi, affects up to 20 million individuals primarily in the Americas where the insect vectors are present and where zoonotic transmission cycles guarantee a steady source of parasites. Increasing travel and immigration has brought T. cruzi infection into the spotlight globally, even in areas where transmission has previously been absent or very low. In these latter situations, congenital and transfusion/transplantation-related transmissions are becoming recognized as significant threats [2,3,4]. A ‘‘conclusive’’ diagnosis of T. cruzi infection is often reached only after multiple serological tests and in combination with epidemiological data and (occasionally) clinical symptoms. Diagnosis of Trypanosoma cruzi infection by direct pathogen detection is complicated by the low parasite burden in subjects persistently infected with this agent of human Chagas disease. Determination of infection status by serological analysis has been faulty, largely due to the lack of well-characterized parasite reagents for the detection of anti-parasite antibodies
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