Design of a piezoelectric immunosensor for tuberculosis biomarker detection

Abstract

Monoclonal antibodies have been applied in biomedical research for diagnosis of various diseases and for the treatment of increasing prevalence diseases such as tuberculosis. There is a need to develop systems capable of detecting tuberculosis-related biomarkers that can provide a quick diagnosis to begin treatment immediately. Thanks to the advances in research for detection, biosensors technologies are emerging as a good alternative for clinical applications. The aim of this work was to develop a piezoelectric immunosensor for rapid and efficient tuberculosis detection. To this purpose, 12 hybridomas and their secreted monoclonal antibodies specific for a biomarker of Mycobacterium tuberculosis were obtained as potential bio-receptors. The monoclonal antibody affinity to the antigenic protein of interest was tested by ELISA. Surface functionalization of the gold electrode -the transducer -of a Quartz Crystal Microbalance (QCM) was simulated by the formation of Self-Assembled Monolayers (SAM) of BSA as a model protein. The functionalized surface was image-characterized by Atomic Force Microscopy (AFM). The appropriate electronic instrumentation system for QCM characterization, with temperature control, was also developed. The obtained immunoreagents will be used as specific recognition bioactive molecules, in combination with functionalized quartz crystals, to build a piezoelectric immunosensor for the detection of Mycobacterium tuberculosis biomarkers, which can be related to the disease.

WITHDRAWN: Bio-sensored Diagnostic Kit for Detecting Tuberculosis Using Artificial Intelligence Through Neural Networks

“…Three steps were followed in this paper such as In Ziehl-Neelsen staining approach after satin image are taken for bacilli for samples and cluster method like K-means is used for extracting tissue and bacilli of tuberculosis and at last features with geometric are considered. This paper experimentally produced result with is more effective and precise to identify the bacilli with tuberculosis in the tissue [5].…”

Mycobacterium tuberculosis 38 kDa Antigen Purification and Potential Diagnostic Use by Piezoelectric Immunosensors

“…Actualmente otro inmunosensor piezoeléctrico para la detección de un biomarcador de TB está siendo desarrollado en Colombia por Jaramillo et al, (2013) empleando un formato directo competitivo. En esta propuesta se inmovilizó antígeno 38kDa recombinante de Mtb sobre la superficie del electrodo de oro de un transductor piezoeléctrico empleando SAMs.…”

“…La estandarización de los parámetros de configuración requeridos, para la caracterización topográfica de superficies de oro biofuncionalizadas con SAM de proteínas mediante AFM, se llevó a cabo realizando estudios de caracterización previa empleando cristales de cuarzo biofuncionalizados con: Albumina de suero bovino (BSA) (Jaramillo et al, 2013), el conjugado del hapteno TCP, BSA-TS1 (Manclús y Montoya, 1996) y CFP de Mtb. En la Tabla 1 se resumen los mejores parámetros de configuración encontrados para un microscopio de fuerza atómica y las características que debe tener la sonda, para la exploración de superficies de oro biofuncionalizadas con antígenos proteicos mediante SAMs.…”

“…Un dispositivo de estas características, que combine las ventajas de un inmunoensayo con la fácil implementación de los biosensores, podría constituirse como un método alternativo para el diagnóstico de TB, facilitando la detección temprana y favoreciendo el tratamiento eficaz. (Montoya et al, 2008;Jaramillo et al, 2013) En este trabajo se presenta el proceso de purificación del antígeno 38kDa nativo (Ag38kDa) a partir de proteínas de secreción filtradas de cultivo (CFP) de Mtb. Tanto las proteínas de secreción filtradas de cultivo, como el antígeno purificado, se emplearon para la inmovilización de cristales piezoeléctricos.…”

The phenomenon of fluorescence in immunosensors.

“…This type of sensors was used for detecting antigens of Mycobacterium tuberculosis (Kumar, 2000;Jaramillo et al, 2013).…”