An Improved Method for the Sensitive Detection of Shiga Toxin 2 in Human Serum
Abstract
:1. Introduction
2. Results and Discussion
2.1. Detection of Stx1 and Stx2 in Stx-Spiked Human Serum by ELISA
2.2. Effect of GuCl on Detection of Stx2 in Serum
2.3. Detection of Stx in Sera of STEC-Infected Patients
2.4. HuSAP Binds Specifically to the Stx2 in Human Serum Samples.
3. Conclusions
4. Materials and Methods
4.1. Stx and Antibodies Used
4.2. Human Serum Samples
4.3. ELISAs for Detection of Stx1 and Stx2
4.4. Co-Immunoprecipitation Assays
4.5. Polyacrylamide Gel Electrophoresis and Western Blot
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Stx Added (ng/mL) | 2 | 4 | 6 | 8 | 10 | Mean ± SD |
---|---|---|---|---|---|---|
Stx1 recovery (%) ± SD | 79.8 ± 11.1 | 74.6 ± 0.8 | 82.5 ± 6.2 | 81.7 ± 4.9 | 86.9 ± 1.7 | 81.1 ± 4.9 |
Stx2 recovery (%) ± SD | 4.8 ± 0.8 | 10.7 ± 0.8 | 17.4 ± 1.4 | 23.3 ± 0.6 | 30.4 ± 0.2 | 17.3 ± 0.7 |
Stx2 Added (ng/mL) | 2 | 4 | 6 | 8 | 10 | Mean ± SD |
---|---|---|---|---|---|---|
Recovery (%) ± SD | 41.23 ± 5.31 | 58.21 ± 1.04 | 66.90 ± 5.21 | 75.72 ± 4.66 | 68.71 ± 3.53 | 62.15 ± 3.95 |
Pt. | Age (y) | Sex | Clinical Symptoms | E. coli Serotype a | Detection of stx Genes in Feces | Stx2 (ng/mL) |
---|---|---|---|---|---|---|
1 | 5.3 | F | Diarrhea | nd b | stx2+ by PCR-based rev dot blot, RT-PCR | 2.99 ± 0.20 |
2 | 0.2 | M | Bloody diarrhea | O157 | stx2+ by PCR-based rev dot blot | 0.40 ± 0.02 |
3 | 0.8 | M | diarrhea, proteinuria | O157 | stx2+ by PCR-based rev dot blot | 4.89 ± 0.77 |
4 | 5.6 | M | diarrhea, proteinuria | O127 | stx2+ by PCR-based rev dot blot, RT-PCR | 3.50 ± 0.75 |
5 | 6.3 | M | Proteinuria | O127 | stx2+ by RT-PCR | 3.57 ± 0.04 |
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He, X.; Ardissino, G.; Patfield, S.; Cheng, L.W.; Silva, C.J.; Brigotti, M. An Improved Method for the Sensitive Detection of Shiga Toxin 2 in Human Serum. Toxins 2018, 10, 59. https://doi.org/10.3390/toxins10020059
He X, Ardissino G, Patfield S, Cheng LW, Silva CJ, Brigotti M. An Improved Method for the Sensitive Detection of Shiga Toxin 2 in Human Serum. Toxins. 2018; 10(2):59. https://doi.org/10.3390/toxins10020059
Chicago/Turabian StyleHe, Xiaohua, Gianluigi Ardissino, Stephanie Patfield, Luisa W. Cheng, Christopher J. Silva, and Maurizio Brigotti. 2018. "An Improved Method for the Sensitive Detection of Shiga Toxin 2 in Human Serum" Toxins 10, no. 2: 59. https://doi.org/10.3390/toxins10020059
APA StyleHe, X., Ardissino, G., Patfield, S., Cheng, L. W., Silva, C. J., & Brigotti, M. (2018). An Improved Method for the Sensitive Detection of Shiga Toxin 2 in Human Serum. Toxins, 10(2), 59. https://doi.org/10.3390/toxins10020059