Journal
BIOSENSORS & BIOELECTRONICS
Volume 78, Issue -, Pages 174-180Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2015.11.041
Keywords
Staphylococcus aureus; Vancomycin; Magnetic beads; Chemiluminescence; Rabbit immunoglobulin G
Categories
Funding
- National Natural Science Foundation of China [21475107]
- Program for Innovative Research Team in University of Chongqing
- Fundamental Research Funds for the Central Universities [XDJK2013A025]
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Vancomycin, which has a strong antibacterial effect to Gram-positive bacteria, was adopted as one molecular recognition agent for bacterial detection. Magnetic beads (MBs) were functionalized with this antibiotic to effectively concentrate Staphylococcus aureus (S. aureus). In addition, alkaline phosphatase (ALP)-tagged rabbit immunoglobulin G (ALP-IgG) was used as the second recognition agent to improve the specificity based on the binding between the Fc region of rabbit IgG and protein A in the cell wall of S. aureus. MBs-concentrated sandwich complex of vancomycin/S. aureus/ALP-IgG was formed with a one-step incubation protocol. Then ALP chemiluminescent reaction was triggered by injecting substrate solution to quantitate S. aureus. Based on the sandwich molecular recognition mechanism and MBs concentration, an ultrasensitive, specific and rapid method was developed for S. aureus detection. The linear range for S. aureus detection was 12-1.2 x 10(6) CFU mL(-1), with a very low detection limit of 3.3 CFU mL(-1). The whole detection process could be completed in 75 min. Other Gram-positive bacteria and Gram-negative bacteria, including Escherichia coli, Salmonella, Pseudomonas aeruginosa, Micrococcus luteus, Bacillus cereus and Bacillus subtilis, showed negligible interference to S. aureus detection. This method was successfully used to quantitate S. aureus in lake water, milk, human urine and human saliva with acceptable recoveries ranging from 70.0% to 116.7%. (C) 2015 Elsevier B.V. All rights reserved.
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