Lateral flow assay-based bacterial detection using engineered cell wall binding domains of a phage endolysin

The development of a cost-effective and efficient bacterial detection assay is essential for diagnostic fields, particularly in resource-poor settings. Although antibodies have been widely used for bacterial capture, the production of soluble antibodies is still expensive and time-consuming. Here, we developed a nitrocellulose-based lateral flow assay using cell wall binding domains (CBDs) from phage as a recognition element and colloidal gold nanoparticles as a colorimetric signal for the detection of a model pathogenic bacterium, Bacillus cereus (B. cereus). To improve conjugation efficiency and detection sensitivity, cysteine-glutathione-S-transferase-tagged CBDs and maltose-binding protein-tagged CBDs were produced in Escherichia coli (E. coli) and incorporated in our assays. The sensitivity of the strip to detect B. cereus was 1x10(4) CFU/mL and the overall assay time was 20 min. The assay showed superior results compared to the antibody-based approach, and did not show any significant cross-reactivity. This proof of concept study indicates that the lateral flow assay using engineered CBDs hold considerable promise as simple, rapid, and cost-effective biosensors for whole cell detection.
Publisher
ELSEVIER ADVANCED TECHNOLOGY
Issue Date
2017-10
Language
English
Keywords

BACILLUS-CEREUS; LISTERIA-MONOCYTOGENES; RAPID DETECTION; ANTIBODIES; EXPRESSION

Citation

BIOSENSORS & BIOELECTRONICS, v.96, pp.173 - 177

ISSN
0956-5663
DOI
10.1016/j.bios.2017.05.010
URI
http://hdl.handle.net/10203/224769
Appears in Collection
BiS-Journal Papers(저널논문)
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