Rapid and sensitive detection of Staphylococcus aureus using biolayer interferometry technology combined with phage lysin LysGH15

Abstract

Staphylococcus aureus (S. aureus), considered as a common foodborne pathogenic microorganism, usually causes food poisoning and various infectious diseases. Therefore, development of rapid and accurate bacterial detection method is the key to preventing food poisoning and achieving early diagnosis and treatment of various infectious diseases caused by S. aureus. Biolayer interferometry (BLI) technology is a novel technique of label-free optical analysis for real-time monitoring of biomolecular interactions. The C54A mutation induced the lytic activity loss of phage lysin LysGH15 but retained the capacity for specific recognizing and binding S. aureus. In this study, a novel method for the detection of S. aureus was established using the C54A mutant LysGH15 as the receptor in combination with BLI. Using this BLI-based method, S. aureus whole cells could be directly assayed and the limit of detection was 13 CFU/mL with a binding time of 12 min. Because the C54A mutant LysGH15 recognizes S. aureus with very high specificity, the method can exclude potential interference from other bacterial species. In addition, this method could also distinguish between viable and dead S. aureus. Moreover, S. aureus was successfully detected in ice cubes and light soy sauce by using this method. Collectively, these results indicate that the LysGH15-based BLI method can be used as an efficient and reliable diagnostic tool in the field of food safety and other related fields for the rapid, sensitive, label-free, and real-time detection of S. aureus.

Keywords: Biolayer interferometry; Biosensor; LysGH15; Phage lysin; Rapid detection; Staphylococcus aureus.