An electrochemical aptasensor for detection of bovine interferon gamma

Bovine tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (MTB) in which it is hard to identify the pathological symptoms. Release of bovine interferon gamma (BoIFN-γ) by T-cells provides an important diagnostic marker of MTB infections. In this work, we developed for the first time an electrochemical aptasensor for sensitive and specific determination of BoIFN-γ. A thiolated IFN-γ-binding aptamer was conjugated with methylene blue (MB) and immobilized on a gold electrode by self-assembly. Binding of IFN-γ to the electrode surface caused a conformational change in the aptamer, decreasing electron transfer efficiency. The redox current was quantified using square wave voltammetry (SWV) and was found to be specific for bovine IFN-γ with detection limit of 0.1 nM in pristine buffer and 0.9 nM in blood. The biosensor described here may, in the future, be used for on-site testing of bovine blood to help better identify and contain outbreaks of bovine TB.

[1]  Kevin W Plaxco,et al.  Aptamer-based electrochemical detection of picomolar platelet-derived growth factor directly in blood serum. , 2007, Analytical chemistry.

[2]  Kevin W Plaxco,et al.  Preparation of electrode-immobilized, redox-modified oligonucleotides for electrochemical DNA and aptamer-based sensing , 2007, Nature Protocols.

[3]  P. Wood,et al.  Development of a simple, rapid in vitro cellular assay for bovine tuberculosis based on the production of gamma interferon. , 1990, Research in veterinary science.

[4]  P. Andersen,et al.  Specific immune-based diagnosis of tuberculosis , 2000, The Lancet.

[5]  S. Jones,et al.  A sandwich enzyme immunoassay for bovine interferon-gamma and its use for the detection of tuberculosis in cattle. , 1990, Australian veterinary journal.

[6]  M. Farrar,et al.  The molecular cell biology of interferon-gamma and its receptor. , 1993, Annual review of immunology.

[7]  R. Potyrailo,et al.  Towards maintenance-free biosensors for hundreds of bind/release cycles. , 2015, Angewandte Chemie.

[8]  M. Denis,et al.  IFN‐γ enhances bovine macrophage responsiveness to Mycobacterium bovis: Impact on bacterial replication, cytokine release and macrophage apoptosis , 2005, Immunology and cell biology.

[9]  S. Downs,et al.  Factors associated with bovine tuberculosis confirmation rates in suspect lesions found in cattle at routine slaughter in Great Britain, 2003-2008. , 2013, Preventive veterinary medicine.

[10]  M. Domingo,et al.  Pathology of bovine tuberculosis. , 2014, Research in veterinary science.

[11]  Zimple Matharu,et al.  Detecting Transforming Growth Factor-β Release from Liver Cells Using an Aptasensor Integrated with Microfluidics , 2014, Analytical chemistry.

[12]  D V Cousins,et al.  Mycobacterium bovis infection and control in domestic livestock. , 2001, Revue scientifique et technique.

[13]  Ian A Gardner,et al.  Consensus recommendations on diagnostic testing for the detection of paratuberculosis in cattle in the United States. , 2006, Journal of the American Veterinary Medical Association.

[14]  Ying Liu,et al.  Aptamer-based electrochemical biosensor for interferon gamma detection. , 2010, Analytical chemistry.

[15]  Ciara K O'Sullivan,et al.  Reagentless, reusable, ultrasensitive electrochemical molecular beacon aptasensor. , 2006, Journal of the American Chemical Society.

[16]  A. Billiau,et al.  Interferon-gamma: biology and role in pathogenesis. , 1996, Advances in immunology.

[17]  R. Juste,et al.  Specific Antibody and Interferon-Gamma Responses Associated with Immunopathological Forms of Bovine Paratuberculosis in Slaughtered Friesian Cattle , 2013, PloS one.

[18]  J. Letesson,et al.  Analysis of the antigen-specific IFN-gamma producing T-cell subsets in cattle experimentally infected with Mycobacterium bovis. , 2002, Veterinary immunology and immunopathology.

[19]  A. Heeger,et al.  Label-free electronic detection of thrombin in blood serum by using an aptamer-based sensor. , 2005, Angewandte Chemie.

[20]  Tanyu Wang,et al.  Quantitation of femtomolar protein levels via direct readout with the electrochemical proximity assay. , 2012, Journal of the American Chemical Society.

[21]  C. Daborn,et al.  The epidemiology of Mycobacterium bovis infections in animals and man: a review. , 1995, Tubercle and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease.