Saltar al contenido
Merck

Fiber-optic immunosensor for detection of Crimean-Congo hemorrhagic fever IgG antibodies in patients.

Analytical chemistry (2015-07-08)
Fairoz Algaar, Evgeni Eltzov, Marina M Vdovenko, Ivan Yu Sakharov, Luka Fajs, Manfred Weidmann, Ali Mirazimi, Robert S Marks
RESUMEN

Crimean-Congo hemorrhagic fever (CCHF) is a severe viral disease with high fatality rate. CCHF virus is endemic in parts of Africa, Asia, the Middle East, and southeastern Europe. Rapid diagnostics of CCHF is vital for appropriate clinical management and prevention of secondary spread from human-to-human. Currently, diagnostics relies on real-time RT-PCR and antibody or antigen detection using ELISA. These methods require trained personnel and expensive equipment and are not appropriate for point-of-care (POC) diagnostics. Furthermore, there are no POC assays available for CCHF. We developed a fiber-optic biosensor for the detection of CCHF IgG antibodies. In order to improve sensitivity, we optimized both the bioreceptor immobilization protocol and the chemiluminescence substrate formulation. The resulting protocol showed a 100-fold greater sensitivity for detection of CCHF antibodies. Finally, we evaluated the fiber-optic biosensor with two CCHF patient sera. We showed that the fiber-optic biosensor is 10-times more sensitive than colorimetric ELISA and is able to detect both patients with high and low levels of IgG antibodies. We believe that the fiber-optic biosensor is a suitable alternative to ELISA as it is much more sensitive and makes it possible to detect a small amount of antibodies at an early stage of infection and can be integrated as a point-of-care diagnostic system of CCHF.

MATERIALES
Referencia del producto
Marca
Descripción del producto

Sigma-Aldrich
Peróxido de hidrógeno solution, 30 % (w/w) in H2O, contains stabilizer
Sigma-Aldrich
Sistema de sustrato líquido 3,3′,5,5′-tetrametilbencidina (TMB) para ELISA, peroxidase substrate
Sigma-Aldrich
Glicina, ReagentPlus®, ≥99% (HPLC)
Sigma-Aldrich
Glicina, suitable for electrophoresis, ≥99%
Sigma-Aldrich
Ácido clorhídrico solution, 1.0 N, BioReagent, suitable for cell culture
Sigma-Aldrich
Ácido sulfúrico, 99.999%
Sigma-Aldrich
Cianoborohidruro de sodio, reagent grade, 95%
Sigma-Aldrich
Glicina, BioUltra, for molecular biology, ≥99.0% (NT)
Sigma-Aldrich
Ácido clorhídrico, 36.5-38.0%, BioReagent, for molecular biology
Sigma-Aldrich
(3-Glicidiloxipropil)trimetoxisilano, ≥98%
Sigma-Aldrich
TWEEN® 20, BioXtra, viscous liquid
Sigma-Aldrich
3,3′,5,5′-Tetrametilbencidina, ≥99%
Supelco
Ácido clorhídrico solution, volumetric, 0.1 M HCl (0.1N), endotoxin free
SAFC
Glicina
Sigma-Aldrich
Glicina, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, ≥98.5%
Sigma-Aldrich
Luminol, 97%
Sigma-Aldrich
Ácido acético, for luminescence, BioUltra, ≥99.5% (GC)
Sigma-Aldrich
Ácido acético, ≥99.5%, FCC, FG