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Refining brucellosis diagnosis by blood transcriptional profiling.

ABSTRACT: Diagnosis of brucellosis remains challenging for several reasons, including lack of culture sensitivity, nonspecific symptomatology, and high prevalence of positive serology in endemic areas. The main objectives of this study were to identify blood biomarkers specific to brucellosis compared to other endemic infections and to monitor changes in blood biomarkers during treatment. To obtain a global profile of the disease, we employed RNA sequencing (RNAseq) of whole blood RNA to measure host response against brucellosis infection in patients from Macedonia and Spain. Long-term follow up of patients was used to classify patients as having acute or chronic/reinfection brucellosis and as treatment responders and non-responders. We observed distinct gene expression differences between samples from acute brucellosis and control donors. The magnitude of gene expression changes was associated with antibody titers determined by standard serological tests for brucellosis, including the rose Bengal and standard agglutination tests. The expression signature characteristic of acute brucellosis was also different from that of subjects with leishmaniasis. In depth integration of clinical data and serological findings with our gene expression data will be performed to provide insight into cellular and molecular mechanisms of brucellosis infection.

ORGANISM(S): Homo sapiens

PROVIDER: GSE69597 | GEO | 2015/06/06

SECONDARY ACCESSION(S): PRJNA285953

REPOSITORIES: GEO

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Project description:Leptospirosis is the most widespread zoonotic disease in the world. The lack of an adequate laboratory test is a major barrier for the diagnosis, especially during the early stages of illness, when antibiotic therapy is most effective. Therefore, there is a critical need for an efficient diagnostic test for this life threatening disease. In order to identify new targets that could be used as diagnostic makers for leptopirosis, we constructed a protein microarray chip comprising 61% of Leptospira interrogans proteome and investigated the IgG response against leptospiral antigens from 274 individuals, including 80 acute-, 80 convalescent-phase patients and 114 healthy control subjects from regions with endemic, high endemic and no endemic transmission of leptospirosis. A nitrocellulose line blot assay was performed to validate the accuracy of the protein microarray results. We found 16 antigens that can discriminate between acute cases and healthy individuals from a region with high endemic transmission of leptospirosis, and 18 antigens that distinguish convalescent cases. Some of the antigens identified in this study, such as LipL32, the non-identical domains of the Lig proteins, GroEL and Loa22 are already known to be recognized by sera from human patients, thus serving as a proof-of-concept for the serodiagnostic antigen discovery approach. Several novel antigens were identified, including the hypothetical protein LIC10215 which showed good sensitivity and specificity rates for both acute- and convalescent-phase patients. Our study is the first large-scale evaluation of immunodominant antigens associated with naturally acquired leptospiral infection and novel as well as known serodiagnostic leptospiral antigens that are recognized by antibodies in the sera of leptospirosis cases were identified. The novel antigens identified here may have potential use in both the development of new tests and the improvement of currently available assays for diagnosing this neglected tropical disease. Further research is needed to assess the accuracy of these antigens in more appropriate diagnostic platforms. Antibody profiling was peformed on sera from infected and non-infected subjects to investigate the IgG response against leptospiral antigens. These samples comprised 80 acute-, 80 convalescent-phase patients and 114 healthy control subjects, including 29 subjects from United States (non-endemic area), 35 blood donors from Salvador/Brazil (endemic area) and 50 healthy individuals from region with high endemicity of leptospirosis.

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Refining brucellosis diagnosis by blood transcriptional profiling.

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