Designing an immunosensor for detection of Brucella abortus based on coloured silica nanoparticles

Abstract

Brucellosis has always been a threat to the health and economics of societies. We report a new colorimetric immunoassay based on colored silica nanoparticles for detection of Brucella abortus. An immunosensor was designed based on blue-SiNPs and paramagnetic nanoparticles (PMNPs). The synthesized immunosensor was conjugated with a polyclonal antibody against B. abortus, which was activated by 1-Ethyl-3–(3-dimethylaminopropyl)-carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to form detection and capture probes, respectively. After adding the conjugates to the bacterial suspension, sandwich structure of PMNPs B. abortus-blue-SiNPs was formed and then separated by a magnet. The blue dye was released from the silica structure and its absorbance was measured at 670 nm with a spectrophotometer. Under optimal conditions, results showed a wide dynamic range from 1.5 × 103 to 1.5 × 108 cfu mL−1 with a detection limit of 450 cfu mL−1. The specificity of the sensor was confirmed in comparison with 5 other bacteria. Also, during the 120-days period, the complex was stable. The results suggested that it can be used in real samples (R2 = .9865). This designed colorimetric immunoassay strategy can be used as an alternative, user-friendly and on-site tool for the rapid detection of Brucella spp. compared to other common methods with high sensitivity and specificity in a short time.

Keywords

• Brucella abortus
• colorimetric
• immunosensor
• nanoparticles
• silica