Location: Quality and Safety Assessment Research UnitTitle: Hyperspectral microscope imaging methods for multiplex detection of Campylobacter
|OAKLEY, BRIAN - Western University Of Health Sciences|
Submitted to: Journal of Spectral Imaging
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/6/2019
Publication Date: 3/10/2019
Citation: Park, B., Eady, M.B., Oakley, B., Yoon, S.C., Lawrence, K.C., Gamble, G.R. 2019. Hyperspectral microscope imaging methods for multiplex detection of Campylobacter. Journal of Spectral Imaging. 8:a6. https://doi.org/10.1255/jsi.2019.a6.
Interpretive Summary: Campylobacter are the major group of bacteria responsible for foodborne gastroenteritis in humans worldwide. More specifically, Campylobacter jejuni is the most common cause of bacterial foodborne illness in the United States. Current method for isolation and detection of Campylobacter from foods is immunological and molecular techniques as well as culture-based with several selective agars designed to isolate Campylobacter colonies. However, these techniques are labor intensive and demand advanced instruments with specially trained experts for the results. Recently, hyperspectral microscope imaging (HMI) technique demonstrated the potential to differentiate and classify foodborne pathogenic live bacteria at the cellular level. The objective of this study is to develop a rapid method for multiplex detection of Campylobacter jejuni using acousto-optic tunable filter (AOTF)-based HMI technique in conjunction with fluorescence in situ hybridization (FISH) technique with high sensitivity and specificity.
Technical Abstract: Campylobacter is an emerging zoonotic bacterial threat in the poultry industry. Current method for isolation and detection of Campylobacter is culture-based techniques with several selective agars designed to isolate Campylobacter colonies. Most significant drawbacks include the time-consuming, labor intensive and have lower sensitivity. Several immunological and molecular techniques such as enzyme-linked immunosorbent assay (ELISA) and Latex agglutination are commercially available for the detection and identification of Campylobacter. However, these methods demand more advanced instruments as well as specially trained experts. A hyperspectral microscope imaging (HMI) system with fluorescence in-situ hybridization (FISH) technique has the potential for multiplex foodborne pathogen detection. Using Alexa488 and Cy3 fluorophores, the HMI technique was able to identify Campylobacter jejuni stains with high sensitivity and specificity. In addition, HMI was able to classify six bacteria with scattering intensity form their spectra without FISH fluorophore. Overall classification accuracy of quadratic discriminant analysis (QDA) method for six bacteria including Bifidobacter longum, Campylobacter jejuni, Clostridium perfringens, Enterobacter cloacae, Lactobacillus salivarius, and Shigella flexneri using a HMI technique without fluorescence markers was 99.4% at the cellular level.