Multicolor and Ultrasensitive Enzyme-Linked Immunosorbent Assay Based on the Fluorescence Hybrid Chain Reaction for Simultaneous Detection of Pathogens.
Xi LvYanmei HuangDaofeng LiuChengwei LiuShan ShanGuoqiang LiMiaolin DuanWei-Hua LaiPublished in: Journal of agricultural and food chemistry (2019)
Various pathogens may coexist in one sample; however, detection methods that rely on traditional selective culture media or immune agents designed specifically for a certain target are unsuitable for multiple targets. It is important to develop a simultaneous and sensitive detection method for multiple pathogens. Here, a multicolor and ultrasensitive enzyme-linked immunosorbent assay (ELISA) platform based on the fluorescence hybridization chain reaction (HCR) was developed. In the assay, multicolor fluorescence concatemers formed as signal amplifiers and signal reporters in the presence of target pathogens. When HCR occurred, Escherichia coli O157:H7, Salmonella serotype Choleraesuis, and Listeria monocytogenes were detected simultaneously with three different fluorescences. Additionally, the limits of detection for E. coli O157:H7, Salmonella Choleraesuis, and L. monocytogenes were 3.4 × 101, 6.4 × 100, and 7.0 × 101 CFU/mL, respectively. The assay achieved ultrasensitive, specific, and simultaneous detection of three pathogens and can be applied to the detection of pathogens in milk samples. Therefore, this multicolor and ultrasensitive ELISA platform has great potential in the application of simultaneous detection of pathogens.
Keyphrases
- label free
- loop mediated isothermal amplification
- escherichia coli
- gram negative
- high throughput
- sensitive detection
- quantum dots
- antimicrobial resistance
- real time pcr
- listeria monocytogenes
- gold nanoparticles
- single molecule
- flow cytometry
- multidrug resistant
- risk assessment
- climate change
- molecularly imprinted
- high resolution
- staphylococcus aureus
- zika virus