Small ubiquitin-related modifier-fused bacteriophage tail fiber protein with favorable aqueous solubility for lateral flow assay of Pseudomonas aeruginosa

Abstract

Rapid and sensitive assay of pathogenic bacteria is critical for minimizing the risk of infectious diseases. Inspired by the interaction between bacteriophages and host bacteria, we obtained a gene sequence of tail fiber protein (TFP) from Pseudomonas aeruginosa (P. aeruginosa) bacteriophage. Then the gene sequence was used to express a recombinant TFP, which can act as a potential capture molecule for P. aeruginosa. Small ubiquitin-related modifier (SUMO) tag was fused onto the TFP fragment to overcome its unfavorable aqueous solubility. The obtained SUMO tag-fused TFP (STFP) can be uniformly distributed onto a nitrocellulose membrane to form a test line due to the improved aqueous solubility, which facilities fabrication of a lateral flow assay strip. Thus we developed a lateral flow assay method by using STFP as a capture molecule and AuCo nanoparticles-labeled aptamer as a signal tracer for point-of-care testing of P. aeruginosa. By using the test strip, P. aeruginosa can be semi quantified with color band and quantified with chemiluminescent (CL) signal catalyzed by AuCo nanoparticles. The concentration range for quantification is 3.3 × 102 CFU/mL to 3.3 × 107 CFU/mL. The test strip was applied to assay P. aeruginosa in different sample matrixes including cerebrospinal fluid, physiological salt solution, drinking water and pear juice. The results demonstrate the application potential of the STFP-based lateral flow assay for medical diagnosis, food and drug safety monitoring.