Lytic phage as a specific and selective probe for detection of Staphylococcus aureus--A surface plasmon resonance spectroscopic study.
Rapid and reliable detection of harmful pathogens at low levels are vital due to the related environmental and economical impact. While antibodies (monoclonal or polyclonal) are successfully employed in many immunoanalysis procedures as a biorecognition element, many of them remain costly with a comparatively short shelf life and uncertain manufacturability. Additionally, they suffer from several limitations, such as susceptibility to hostile environmental stresses such as temperature, pH, ionic strength, and cross-reactivity. The development of easy available, sensitive, and robust alternative molecular recognition elements, capable of providing a very high level of selectivity are very attractive to industry and may benefit in multiple areas. Several attempts have been made to utilize fluorescent-tagged bacteriophages and phage-displayed peptides for bacterial detection. However, involvement of complex labeling and detecting procedures make these approaches time-consuming and complicated. Here, we are reporting for the first time, the label-free detection of Staphylococcus aureus using lytic phage as highly specific and selective biorecognition element and surface plasmon resonance-based SPREETA sensor as a detection platform. Lytic phage was immobilized on the gold surface of SPREETA sensor via trouble-free direct physical adsorption. The detection limit was found to be 10(4) cfu/ml. Detection specificity was investigated by an inhibition assay while selectivity was examined with Salmonella typhimurium. The preliminary results using lytic phage as a probe for bacterial detection, in combination with SPR platform are promising and hence can be employed for rapid and label-free detection of different bacterial pathogens.
Balasubramanian
Shankar
S
Materials Research and Education Center, Samuel Ginn College of Engineering, Auburn University, 275 Wilmore, Auburn, AL 36849, USA.
Sorokulova
Iryna B
IB
Vodyanoy
Vitaly J
VJ
Simonian
Aleksandr L
AL
D001435:Bacteriophages / Q000302:isolation & purification* / Q000502:physiology*; D001681:Biological Assay / Q000379:methods*; D015374:Biosensing Techniques / Q000379:methods*; D015169:Colony Count, Microbial / Q000379:methods*; D015336:Molecular Probe Techniques; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013211:Staphylococcus aureus / Q000302:isolation & purification* / Q000821:virology; D020349:Surface Plasmon Resonance / Q000379:methods*