Diagnostic Performance of Tuberculosis-Specific IgG Antibody Profiles in Patients with Presumptive Tuberculosis from Two Continents.

BACKGROUND: Development of rapid diagnostic tests for tuberculosis is a global priority. A whole proteome screen identified Mycobacterium tuberculosis antigens associated with serological responses in tuberculosis patients. We used World Health Organization (WHO) target product profile (TPP) criteria for a detection test and triage test to evaluate these antigens. METHODS: Consecutive patients presenting to microscopy centers and district hospitals in Peru and to outpatient clinics at a tuberculosis reference center in Vietnam were recruited. We tested blood samples from 755 HIV-uninfected adults with presumptive pulmonary tuberculosis to measure IgG antibody responses to 57 M. tuberculosis antigens using a field-based multiplexed serological assay and a 132-antigen bead-based reference assay. We evaluated single antigen performance and models of all possible 3-antigen combinations and multiantigen combinations. RESULTS: Three-antigen and multiantigen models performed similarly and were superior to single antigens. With specificity set at 90% for a detection test, the best sensitivity of a 3-antigen model was 35% (95% confidence interval [CI], 31-40). With sensitivity set at 85% for a triage test, the specificity of the best 3-antigen model was 34% (95% CI, 29-40). The reference assay also did not meet study targets. Antigen performance differed significantly between the study sites for 7/22 of the best-performing antigens. CONCLUSIONS: Although M. tuberculosis antigens were recognized by the IgG response during tuberculosis, no single antigen or multiantigen set performance approached WHO TPP criteria for clinical utility among HIV-uninfected adults with presumed tuberculosis in high-volume, urban settings in tuberculosis-endemic countries.

Current microarray surface chemistries.

In almost all microarray technologies that are currently used, some type of surface chemistry serves as the interface between immobilized biomolecules and the solid support. Factors such as probe loading, spot morphology, and signal-to-noise ratio are all intimately linked to surface chemistry. Surface chemistry also significantly impacts important performance parameters such as three-dimensional structure of the immobilized biomolecules and nonspecific assay backgrounds. Here, an overview of the major types of surface chemistries currently used in printed microarrays is provided, with an emphasis on standard glass slide formats. The first part of this chapter focuses on DNA array surface chemistries, including both commercially fabricated and custom-made arrays. The second part of the chapter focuses on emerging protein, peptide, and carbohydrate array techniques. The intent is to provide the molecular biology researcher and bio-analytical or diagnostic specialist with a guide to the surface chemistry state-of-the-art for established and emerging array technologies.