Surface Plasmon Resonance Immunosensor for the Detection of Campylobacter jejuni

Abstract

Abstract:Campylobacteriosis is an internationally important foodborne disease caused by Campylobacter jejuni. The bacterium is prevalent in chicken meat and it is estimated that as much as 90% of chicken meat on the market may be contaminated with the bacterium. The current gold standard for the detection of C. jejuni is the culturing method, which takes at least 48 h to confirm the presence of the bacterium. Hence, the aim of this work was to investigate the development of a Surface Plasmon Resonance (SPR) sensor platform for C. jejuni detection. Bacterial strains were cultivated in-house and used in the development of the sensor. SPR sensor chips were first functionalized with polyclonal antibodies raised against C. jejuni using covalent attachment. The gold chips were then applied for the direct detection of C. jejuni. The assay conditions were then optimized and the sensor used for C. jejuni detection, achieving a detection limit of 8 × 10 6 CFU·mL −1 . The sensitivity of the assay was further enhanced to 4 × 10 4 CFU·mL −1 through the deployment of a sandwich assay format using the same polyclonal antibody. The LOD obtained in the sandwich assay was higher than that achieved using commercial enzyme-linked immunosorbent assay (ELISA) (10 6 -10 7 CFU·mL −1 ). This indicate that the SPR-based sandwich sensor method has an excellent potential to replace ELISA tests for C. jejuni detection. Specificity studies performed with Gram-positive and Gram-negative bacteria, demonstrated the high specific of the sensor for C. jejuni.

New Colorimetric DNA Sensor for Detection of Campylobacter jejuni in Milk Sample Based on Peroxidase‐Like Activity of Gold/Platinium Nanocluster

“…Other risk factors include consumption of animal products, such as raw milk products, and contaminated water that has been in contact with animals, and international travel . In the last three decades, many researchers have been focusing on development of reliable and quick diagnostic methods in order to decrease the effect of Campylobacter jejuni on the human and animal's health . The gold standard for the tracing of C. jejuni in various samples is the culture technique, which requires special culture media and microaerophilic condition which bacteria growth and formation of seeable colony takes more than 48 h .…”

“…Expire date for Food products such as poultry are commonly a few days after the slaughtering, thus culture method for the fast analysis of Campylobacter jejuni was failed. Therefore, research on the rapid and convenient method centralized . Concurrent with culture method improvement other methods such as enzyme immunoassay, PCR, and Real‐Time PCR applied for Campylobacter jejuni detection .…”

“…The best candidate that can achieve these aims is biosensor technology. Biosensors have many benefits over conventional detection methods such as selectivity, sensitivity, and portability without time‐consuming procedures …”

“…SPR biosensors have advantages such as high sensitivity and real‐time monitoring . SPR method based on sandwich immunoassay and secondary antibody conjugated with gold nanoparticles was applied for Campylobacter jejuni detection . SPR method not only need a gold chip with high price but also require several steps such as surface chemical functionalization and bio‐molecular immobilization .…”

“…[3,4] In the last three decades, many researchers have been focusing on development of reliable and quick diagnostic methods in order to decrease the effect of Campylobacter jejuni on the human and animal's health. [6] The gold standard for the tracing of C. jejuni in various samples is the culture technique, which requires special culture media and microaerophilic condition which bacteria growth and formation of seeable colony takes more than 48 h. [7] Expire date for Food products such as poultry are commonly a few days after the slaughtering, thus culture method for the fast analysis of Campylobacter jejuni was failed. Therefore, research on the rapid and convenient method centralized.…”