He Dongxia,Liu Qi,Wang Lei,Han Qinqin,Zhang Jinyang,Li Chao,Song Yuzhu
A novel strategy integrating Brevinin-1BW (BW)-functionalized magnetic bead-based separation with a colorimetric immunoassay was developed to concentrate and enrich Staphylococcus aureus ( S. aureus ) from sample matrices using magnetic beads functionalized with antimicrobial peptide BW (MBs-BW) as carriers. To ensure the selectivity of the method, horseradish peroxidase (HRP)-labeled porcine IgG (HRP-porcine IgG) was used as a molecular recognition reagent and signal amplification probe. MBs-BW/ S. aureus /HRP-porcine IgG probe sandwich complex was successfully obtained. HRP chromogenic substrate 3,3′,5,5′-tetramethylbenzidine (TMB) was used to realize the chromogenic reaction. Finally, S. aureus was quantitatively analyzed based on the color signal generated. Under the optimal conditions, the assay showed a response in the concentration range of 1.0 × 10 2 –1.0 × 10 7 CFU/mL of S. aureus , and the detection limit was as low as 60 CFU/mL. This method does not require complicated sample pretreatment, and the operation process is fast and simple, which can directly complete the whole process of bacterial enrichment and detection. The recovery of this strategy in different types of spiked samples reached 86.8–97.8%, and the analytical results of this strategy were highly consistent with those of the conventional plate counting method. This strategy provides a new way for the rapid detection of S. aureus in complex samples. Graphical
Zhizhi Chen,Lei Wang,Dongxia He,Qi Liu,Qinqin Han,Jinyang Zhang,A-Mei Zhang,Yuzhu Song
Antibiotic resistance has emerged as a grave threat to global public health, leading to an increasing number of treatment failures. Antimicrobial peptides (AMPs) are widely regarded as potential substitutes for traditional antibiotics since they are less likely to induce resistance when used. A novel AMP named Brevinin-1BW (FLPLLAGLAASFLPTIFCKISRKC) was obtained by the Research Center of Molecular Medicine of Yunnan Province from the skin of thePelophylax nigromaculatus. Brevinia-1BW had effective inhibitory effects on Gram-positive bacteria, with a minimum inhibitory concentration (MIC) of 3.125 μg/mL againstEnterococcus faecalis(ATCC 29212) and 6.25 μg/mL against bothStaphylococcus aureus(ATCC 25923) and multidrug-resistantStaphylococcus aureus(ATCC 29213) but had weaker inhibitory effects on Gram-negative bacteria, with a MIC of ≥100 μg/mL. Studies using scanning electron microscopy (SEM) and flow cytometry have revealed that it exerts its antibacterial activity by disrupting bacterial membranes. Additionally, it possesses strong biofilm inhibitory and eradication activities as well as significant lipopolysaccharide (LPS)-binding activity. Furthermore, Brevinin-1BW has shown a significant anti-inflammatory effect in LPS-treated RAW264.7 cells. In conclusion, Brevinin-1BW is anticipated to be a promising clinical agent with potent anti-Gram-positive bacterial and anti-inflammatory properties.
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