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Lysin cell-binding domain-functionalized magnetic beads for detection of Staphylococcus aureus via inhibition of fluorescence of Amplex Red/hydrogen peroxide assay by intracellular catalase.

Abstract

Accurate and rapid identification of Staphylococcus aureus (S. aureus) is of great significance for controlling the food poisoning and infectious diseases caused by S. aureus. In this study, a novel strategy that combines lysin cell-binding domain (CBD)-based magnetic separation with fluorescence detection was developed for the specific and sensitive quantification of S. aureus in authentic samples. The S. aureus cells were separated from the sample matrix by lysin CBD-functionalized magnetic beads. Following lysis by lysostaphin, intracellular catalase was released from S. aureus cells and detected by a fluorometric system composed of horseradish peroxidase (HRP), hydrogen peroxide (H2O2), and Amplex Red. S. aureus was quantified via the inhibitory effect of the released intracellular catalase on the fluorometric system since the catalase could decompose the H2O2. Optimized conditions afforded a calibration curve for S. aureus ranging from 1.0 × 102 to 1.0 × 107 CFU mL-1. The detection limit was as low as 78 CFU mL-1 in phosphate-buffered saline (PBS), and the total detection process could be completed in less than 50 min. Other bacteria associated with common food-borne and nosocomial infections negligibly interfered with S. aureus detection, except for Staphylococcus epidermidis, which may have slightly interfered. Moreover, the potential of this proposed method for practical applications has been demonstrated by detection assays of sterilized milk and human serum. Graphical abstract.

Authors+Show Affiliations

Department of Medical Laboratory, Weifang Medical University, Weifang, 261053, Shandong, China.Department of Cardiovascular Medicine, Weifang People's Hospital, Weifang, 261041, Shandong, China.Department of Medical Laboratory, Weifang Medical University, Weifang, 261053, Shandong, China.Department of Medical Laboratory, Weifang Medical University, Weifang, 261053, Shandong, China.Department of Medical Laboratory, Weifang Medical University, Weifang, 261053, Shandong, China.Department of Medical Laboratory, Weifang Medical University, Weifang, 261053, Shandong, China.Department of Medical Laboratory, Weifang Medical University, Weifang, 261053, Shandong, China. ronglanzhao@wfmc.edu.cn.Department of Medical Laboratory, Weifang Medical University, Weifang, 261053, Shandong, China. qiaojj@wfmc.edu.cn.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31522243

Citation

Yi, Zhengjun, et al. "Lysin Cell-binding Domain-functionalized Magnetic Beads for Detection of Staphylococcus Aureus Via Inhibition of Fluorescence of Amplex Red/hydrogen Peroxide Assay By Intracellular Catalase." Analytical and Bioanalytical Chemistry, 2019.
Yi Z, Wang S, Meng X, et al. Lysin cell-binding domain-functionalized magnetic beads for detection of Staphylococcus aureus via inhibition of fluorescence of Amplex Red/hydrogen peroxide assay by intracellular catalase. Anal Bioanal Chem. 2019.
Yi, Z., Wang, S., Meng, X., Wu, A., Li, Q., Song, Y., ... Qiao, J. (2019). Lysin cell-binding domain-functionalized magnetic beads for detection of Staphylococcus aureus via inhibition of fluorescence of Amplex Red/hydrogen peroxide assay by intracellular catalase. Analytical and Bioanalytical Chemistry, doi:10.1007/s00216-019-02099-0.
Yi Z, et al. Lysin Cell-binding Domain-functionalized Magnetic Beads for Detection of Staphylococcus Aureus Via Inhibition of Fluorescence of Amplex Red/hydrogen Peroxide Assay By Intracellular Catalase. Anal Bioanal Chem. 2019 Sep 14; PubMed PMID: 31522243.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Lysin cell-binding domain-functionalized magnetic beads for detection of Staphylococcus aureus via inhibition of fluorescence of Amplex Red/hydrogen peroxide assay by intracellular catalase. AU - Yi,Zhengjun, AU - Wang,Shuhui, AU - Meng,Xiangying, AU - Wu,Anqi, AU - Li,Qian, AU - Song,Yongjie, AU - Zhao,Ronglan, AU - Qiao,Jinjuan, Y1 - 2019/09/14/ PY - 2019/06/13/received PY - 2019/08/22/accepted PY - 2019/07/27/revised PY - 2019/9/16/entrez PY - 2019/9/16/pubmed PY - 2019/9/16/medline KW - Amplex Red KW - Catalase KW - Lysin cell-binding domain KW - Magnetic separation KW - Staphylococcus aureus JF - Analytical and bioanalytical chemistry JO - Anal Bioanal Chem N2 - Accurate and rapid identification of Staphylococcus aureus (S. aureus) is of great significance for controlling the food poisoning and infectious diseases caused by S. aureus. In this study, a novel strategy that combines lysin cell-binding domain (CBD)-based magnetic separation with fluorescence detection was developed for the specific and sensitive quantification of S. aureus in authentic samples. The S. aureus cells were separated from the sample matrix by lysin CBD-functionalized magnetic beads. Following lysis by lysostaphin, intracellular catalase was released from S. aureus cells and detected by a fluorometric system composed of horseradish peroxidase (HRP), hydrogen peroxide (H2O2), and Amplex Red. S. aureus was quantified via the inhibitory effect of the released intracellular catalase on the fluorometric system since the catalase could decompose the H2O2. Optimized conditions afforded a calibration curve for S. aureus ranging from 1.0 × 102 to 1.0 × 107 CFU mL-1. The detection limit was as low as 78 CFU mL-1 in phosphate-buffered saline (PBS), and the total detection process could be completed in less than 50 min. Other bacteria associated with common food-borne and nosocomial infections negligibly interfered with S. aureus detection, except for Staphylococcus epidermidis, which may have slightly interfered. Moreover, the potential of this proposed method for practical applications has been demonstrated by detection assays of sterilized milk and human serum. Graphical abstract. SN - 1618-2650 UR - https://www.unboundmedicine.com/medline/citation/31522243/Lysin_cell-binding_domain-functionalized_magnetic_beads_for_detection_of_Staphylococcus_aureus_via_inhibition_of_fluorescence_of_Amplex_Red/hydrogen_peroxide_assay_by_intracellular_catalase L2 - https://dx.doi.org/10.1007/s00216-019-02099-0 DB - PRIME DP - Unbound Medicine ER -