A Highly Sensitive ELISA and Immunochromatographic Strip for the Detection of Salmonella typhimurium in Milk Samples
Abstract
:1. Introduction
2. Materials and Methods
2.1. Strains and Growth Conditions
2.2. Purification and Characterization of S. typhimurium Flagellin
2.3. Monoclonal Antibodies for Detecting S. typhimurium Flagellin and LPS
2.4. The Use of a Combination of Flagellin and LPS MAbs to Develop a Sandwich ELISA for Detecting S. typhimurium
2.5. The Establishment and Cross-Reactivity of LPS mAb-Based Sandwich ELISA
2.6. LPS mAb-Based Immunochromatographic Strip for S. typhimurium
2.7. Milk Samples
3. Results and Discussion
3.1. Characterization of the S. typhimurium Flagellin Preparation
3.2. Characterization of S. typhimurium Flagellin and LPS MAbs
F1 IgG1 | F2 IgG1 | F3 IgG1 | F4 IgG1 | L1 IgG2b | L2 IgG2b | L3 IgG1 | L4 IgG2b | L5 IgM | L6 IgG1 | |
---|---|---|---|---|---|---|---|---|---|---|
S. typhimurium ATCC 13311 | ++ | + | + | + | +++ | +++ | +++ | +++ | +++ | +++ |
S. typhimurium ATCC 14028 | + | + | + | - | +++ | +++ | +++ | +++ | +++ | +++ |
S. paratphi B CMCC 50094 | - | - | - | - | +++ | +++ | +++ | +++ | +++ | +++ |
S. enteritidis ATCC 13076 | - | - | - | - | - | - | - | - | - | - |
LPS | +++ | +++ | +++ | +++ | +++ | +++ | ||||
Ra LPS | - | - | - | - | - | - |
3.3. A Combination of Flagellin and LPS MAbs for the Detection of S. typhimurium
F1 | F2 | F3 | F4 | L1 | L2 | L3 | L4 | L5 | L6 | |
---|---|---|---|---|---|---|---|---|---|---|
F1-HRP | 7.68 | 7.24 | 4.93 | |||||||
F2-HRP | ||||||||||
F3-HRP | 2.28 | |||||||||
F4-HRP | ||||||||||
L1-HRP | 7.72 | 6.37 | 6.93 | 6.42 | 9.62 | 14.18 | 13.88 | 10.03 | 11.55 | 12.45 |
L2-HRP | 14.88 | 9.75 | 19.19 | 14.80 | 15.42 | 14.80 | 17.47 | 17.38 | 10.95 | 15.47 |
L3-HRP | 2.34 | 2.42 | 3.30 | 9.38 | 2.86 | 10.48 | 8.30 | |||
L4-HRP | 10.12 | 7.16 | 10.57 | 7.91 | 14.81 | 14.50 | 16.66 | 15.10 | 17.37 | 15.65 |
L5-HRP | 16.41 | 12.81 | 14.15 | 12.3 | 20.19 | 17.14 | 19.51 | 20.38 | 17.27 | 12.58 |
L6-HRP | 18.35 | 7.81 | 13.83 | 6.47 | 17.63 | 22.66 | 22.37 | 21.00 | 20.50 | 20.09 |
3.4. The Establishment and Cross-Reactivity of the LPS mAb-Based Sandwich ELISA
3.5. LPS MAbs-Based Immunochromatographic Strip for S. typhimurium
3.6. Detection of Milk Samples
Sandwich ELISA | Immunochromatographic | Plate Counting | |
---|---|---|---|
Sensitivity | (1.83 ± 0.32) × 105 | 1.25 × 106 | ≤(1.9 ± 0.32) × 104 |
Detection time | 3 h | 10 min | 24 h |
Specificity | Specific | Specific | - |
Portability | - | Portable | - |
Simplicity of operation | - | Very simple | - |
4. Conclusions
Supplementary Files
Supplementary File 1Acknowledgments
Author Contributions
Conflicts of Interest
References
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Wang, W.; Liu, L.; Song, S.; Tang, L.; Kuang, H.; Xu, C. A Highly Sensitive ELISA and Immunochromatographic Strip for the Detection of Salmonella typhimurium in Milk Samples. Sensors 2015, 15, 5281-5292. https://doi.org/10.3390/s150305281
Wang W, Liu L, Song S, Tang L, Kuang H, Xu C. A Highly Sensitive ELISA and Immunochromatographic Strip for the Detection of Salmonella typhimurium in Milk Samples. Sensors. 2015; 15(3):5281-5292. https://doi.org/10.3390/s150305281
Chicago/Turabian StyleWang, Wenbin, Liqiang Liu, Shanshan Song, Lijuan Tang, Hua Kuang, and Chuanlai Xu. 2015. "A Highly Sensitive ELISA and Immunochromatographic Strip for the Detection of Salmonella typhimurium in Milk Samples" Sensors 15, no. 3: 5281-5292. https://doi.org/10.3390/s150305281
APA StyleWang, W., Liu, L., Song, S., Tang, L., Kuang, H., & Xu, C. (2015). A Highly Sensitive ELISA and Immunochromatographic Strip for the Detection of Salmonella typhimurium in Milk Samples. Sensors, 15(3), 5281-5292. https://doi.org/10.3390/s150305281