Different Assay Conditions for Detecting the Production and Release of Heat-Labile and Heat-Stable Toxins in Enterotoxigenic Escherichia coli Isolates
Abstract
:1. Introduction
2. Results and Discussion
2.1. Characterization of ST MAb
2.2. Comparison of Effects of Different Compounds on Production of LT and ST by E. coli H10407
2.3. Effects of Antibiotics on Toxin Production
2.4. Characterization of LT and ST Production/Release in ETEC Strains
2.5. Discussion
3. Experimental Section
3.1. Bacterial Strains
Strain | Serotype | Gene | Toxin expression | Geographic origin/period of isolation |
---|---|---|---|---|
18 | O8:H9 | elt | (LT+) | São Paulo, Brazil, 1994–1996 |
28 | O112:H10 | elt | (LT+) | São Paulo, Brazil, 1994–1996 |
51 | O62:H19 | elt | (LT+) | São Paulo, Brazil, 1994–1996 |
120 | O88:H25 | elt | (LT+) | São Paulo, Brazil, 1994–1996 |
10/1A | O114:H21 | elt | (LT+) | Rio de Janeiro, Brazil, 1998 |
10/1B | O114:H21 | elt | (LT+) | Rio de Janeiro, Brazil, 1998 |
105A1 | ONT:H4 | elt | (LT+) | Paraiba, Brazil, 2000–2001 |
117A1 | O6:H16 | elt | (LT+) | Paraiba, Brazil, 2000–2001 |
162-1 | O88:H25 | elt | (LT+) | São Paulo, Brazil, 1989–1990 |
18/1 | ONT:HNT | elt | (LT+) | Rio de Janeiro, Brazil, 1998 |
21089 | ONT:HNT | elt | (LT+) | Rio de Janeiro, Brazil, 1998 |
220A1 | O25:H16 | elt | (LT+) | Paraiba, Brazil, 2000–2001 |
258909-3 | O128:H1 | elt | (LT+) | Bangladesh, 1979–1984 |
2A5 | O88:H25 | elt | (LT+) | Paraiba, Brazil, 2000–2001 |
3841-3 | O88:H- | elt | (LT+) | São Paulo, Brazil, 1989–1990 |
39A1 | O109:H19 | elt | (LT+) | Paraiba, Brazil, 2000–2001 |
4541-5 | O64:H- | elt | (LT+) | São Paulo, Brazil, 1989–1990 |
4702-1 | O167:H5 | elt | (LT+) | São Paulo, Brazil, 1989–1990 |
72A1 | O15:H40 | elt | (LT+) | Paraiba, Brazil, 2000–2001 |
102330-1 | O45:H16 | elt | (LT+) | Bahia, Brazil, 2001–2002 |
106172-1 | ONT:H10 | elt | (LT+) | Bahia, Brazil, 2001–2002 |
115181-1 | O64:H- | elt | (LT+) | Bahia, Brazil, 2001–2002 |
117820-1 | O64:H- | elt | (LT+) | Bahia, Brazil, 2001–2002 |
3400-1 | ONT:H4 | elt | (LT+) | Bahia, Brazil, 2001–2002 |
104 | OR:H17 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
138 | OR:H25 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
224 | OR:H10 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
231 | OR:H25 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
308 | OR:H16 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
622 | OR:H9 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
906 | O15:H40 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
913 | O6:H16 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
922 | O64:HNT | elt | (LT+) | Bahia, Brazil, 2003–2004 |
985 | O166:H15 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
1146 | OR:H21 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
1334 | ONT:H16 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
1530 | OR:H19 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
1754 | OR:H21 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
2109 | ONT:H32 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
2335 | OR:H51 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
2464 | OR:H10 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
3081 | OR:H- | elt | (LT+) | Bahia, Brazil, 2003–2004 |
3412 | O109:H- | elt | (LT+) | Bahia, Brazil, 2003–2004 |
3584 | OR:H32 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
3628 | ONT:H40 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
3684 | O133:H25 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
4125 | O82:H40 | elt | (LT+) | Bahia, Brazil, 2003–2004 |
2004 | ONT:H- | elt | (LT+) | Bahia, Brazil, 2003–2004 |
160BA | OR:H- | elt | (LT+) | Bahia, Brazil, 2003–2004 |
3321-4 | O153:H45 | estA | (ST+) | São Paulo, Brazil, 1989–1990 |
616 | O25:H16 | estA | (ST+) | Bahia, Brazil, 2003–2004 |
2016 | O8:H2 | estA | (ST+) | Bahia, Brazil, 2003–2004 |
6546-1 | ONT:H32 | estA | (ST+) | Bahia, Brazil, 2001–2002 |
416 | O148:H27 | estA | (ST+) | Bahia, Brazil, 2003–2004 |
2859 | O78:H27 | estA | (ST+) | Bahia, Brazil, 2003–2004 |
3541 | ONT:HNT | estA | (ST+) | Bahia, Brazil, 2003–2004 |
74499-1 | O23:H15 | estA | (ST+) | Bahia, Brazil, 2003–2004 |
76359-1 | O166:H15 | estA | (ST+) | Bahia, Brazil, 2001–2002 |
1791-1 | O29:H21 | estA | (ST+) | São Paulo, Brazil, 1989–1990 |
4961-2 | O29:H21 | estA | (ST+) | São Paulo, Brazil, 1989–1990 |
501-4 | O78:H12 | estA | (ST+) | São Paulo, Brazil, 1989–1990 |
O211-1 | O6:H16 | estA | (ST+) | São Paulo, Brazil, 1989–1990 |
3231-4 | ONT:H[NT] | estA | (ST+) | São Paulo, Brazil, 1989–1990 |
2021-1 | O128ac:H27 | estA | (ST+) | São Paulo, Brazil, 1989–1990 |
3511-1 | O128ac:H21 | estA | (ST+) | São Paulo, Brazil, 1989–1990 |
4011-1 | O153:H45 | estA | (ST+) | São Paulo, Brazil, 1989–1990 |
3891-1 | O153:H45 | estA | (ST+) | São Paulo, Brazil, 1989–1990 |
75525-1 | O27:H7 | estA | (ST+) | Bahia, Brazil, 2001–2002 |
84/3353 | O4:H1 | estA | (ST+) | Germany, 1984 |
84/3046 | O6:HNT | estA | (ST+) | Germany, 1984 |
84/3610 | ONT:H5 | estA | (ST+) | Germany, 1984 |
H10407 | O78:H11 | elt/estA | (LT+ ST+) | Bangladesh, 1973 |
4 | O6:H16 | elt/estA | (LT+ ST+) | São Paulo, Brazil, 1994–1996 |
5 | O6:H16 | elt/estA | (LT+ ST+) | São Paulo, Brazil, 1994–1996 |
155A1 | O6:H16 | elt/estA | (LT+ ST+) | Paraiba, Brazil, 2000–2001 |
156A1 | O6:H16 | elt/estA | (LT+ ST+) | Paraiba, Brazil, 2000–2001 |
157A2 | O6:H16 | elt/estA | (LT+ ST+) | Paraiba, Brazil, 2000–2001 |
159A2 | O6:H16 | elt/estA | (LT+ ST+) | Paraiba, Brazil, 2000–2001 |
160A2 | O6:H16 | elt/estA | (LT+ ST+) | Paraiba, Brazil, 2000–2001 |
170A1 | O6:H16 | elt/estA | (LT+ ST+) | Paraiba, Brazil, 2000–2001 |
40T | OR:H- | elt/estA | (LT+ ST+) | São Paulo, Brazil, 1989–1990 |
99237 | O78:H12 | elt/estA | (LT+ ST+) | Rio de Janeiro, Brazil, 1998 |
237 | O6:H16 | elt/estA | (LT+ ST+) | Bahia, Brazil, 2003–2004 |
2355 | OR:H16 | elt/estA | (LT+ ST+) | Bahia, Brazil, 2003–2004 |
3026 | O6:H16 | elt/estA | (LT+ ST+) | Bahia, Brazil, 2003–2004 |
3095 | O6:H16 | elt/estA | (LT+ ST+) | Bahia, Brazil, 2003–2004 |
3238 | O6:H16 | elt/estA | (LT+ ST+) | Bahia, Brazil, 2003–2004 |
3950 | O6:H16 | elt/estA | (LT+ ST+) | Bahia, Brazil, 2003–2004 |
15 | ONT:H19 | elt/estA | (LT+ ST+) | São Paulo, Brazil, 1994–1996 |
170 | O6:H16 | elt/estA | (LT+ ST+) | São Paulo, Brazil, 1994–1996 |
3.2. O:H Identification
3.3. Media, Culture Conditions and Treatments for Toxin Release
3.4. Rabbit Polyclonal and Mouse Monoclonal (MAb) Anti-LT Antibodies
3.5. Rabbit Polyclonal and Mouse Monoclonal (MAb) Anti-ST Antibodies
3.6. Evaluation of Anti-ST MAb Reactivity
3.7. Indirect ELISA for ST
3.8. Capture ELISA for ST
3.9. cELISA for LT
3.10. Statistical Analysis
4. Conclusions
Acknowledgments
Conflicts of Interest
References
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Rocha, L.B.; Ozaki, C.Y.; Horton, D.S.P.Q.; Menezes, C.A.; Silva, A.; Fernandes, I.; Magnoli, F.C.; Vaz, T.M.I.; Guth, B.E.C.; Piazza, R.M.F. Different Assay Conditions for Detecting the Production and Release of Heat-Labile and Heat-Stable Toxins in Enterotoxigenic Escherichia coli Isolates. Toxins 2013, 5, 2384-2402. https://doi.org/10.3390/toxins5122384
Rocha LB, Ozaki CY, Horton DSPQ, Menezes CA, Silva A, Fernandes I, Magnoli FC, Vaz TMI, Guth BEC, Piazza RMF. Different Assay Conditions for Detecting the Production and Release of Heat-Labile and Heat-Stable Toxins in Enterotoxigenic Escherichia coli Isolates. Toxins. 2013; 5(12):2384-2402. https://doi.org/10.3390/toxins5122384
Chicago/Turabian StyleRocha, Letícia B., Christiane Y. Ozaki, Denise S. P. Q. Horton, Caroline A. Menezes, Anderson Silva, Irene Fernandes, Fabio C. Magnoli, Tania M. I. Vaz, Beatriz E. C. Guth, and Roxane M. F. Piazza. 2013. "Different Assay Conditions for Detecting the Production and Release of Heat-Labile and Heat-Stable Toxins in Enterotoxigenic Escherichia coli Isolates" Toxins 5, no. 12: 2384-2402. https://doi.org/10.3390/toxins5122384