Table 9.
In vitro methods of BoNT detection in human serum.
| BoNT Type | Method | Serum Samples | Sensitivity | References |
|---|---|---|---|---|
| A | Assay with a large immunosorbent surface area (ALISSA) | spiked human serum | Attomolar (4 × 10−5 MLD/mL) a |
[168,169] |
| A, B, C, D, E, F | Endopeptidase assay and monoclonal neoepitope antibodies | spiked human serum | A: 0.211 MLD50/mL B: 0.147 MLD50/mL C: 2.056 MLD50/mL D: 0.18 MLD50/mL E: 0.84 MLD50/mL F: 0.005 MLD50/mL |
[170] |
| A, B, E, F | Endopep-MS | spiked human serum | A: 20 MLD/mL B, F: 1 MLD/mL E: 0.2 MLD/mL |
[171] |
| A, B, E, F | Endopep-MALDI-TOF-MS | spiked human serum | A, B, F: 0.1 MLD/mL E: 0.02 MLD/mL |
[172,173,174] |
| A, B, F | Endopeptidase assay with fluorogenic substrates | spiked human serum | A, F: 1 pM (A: 40 MLD/mL F: 3 MLD/mL) a B: 10 pM (180 MLD/mL) a |
[175,176] |
| A | Endopep-MS | spiked human serum | 1 MLD/mL | [177,178] |
| A, B | Endopep-MS | spiked macaque serum | 1 MLD/mL | [179] |
| A, B, D/C, E, F | Endopep-MS | spiked human, chicken sera | A, B, F: 1 MLD/mL D/C: 2 MLD/mL E: 10 MLD/mL C: 100 MLD/mL |
[180] |
| C | Endopep-MS | spiked human serum | 0.5–1 MLD/mL | [181] |
| A | Microfluidic double sandwich immunoassay | spiked human serum | 30 pg/mL (8 MLD/mL) a |
[182] |
| A | Functional dual coating assay (BoNT immunocapture and endopeptidase assay with detection by neoepitope antibodies) | clinical serum samples | 1 MLD/mL | [183,184] |
| A | Endopeptidase assay and detection with neoepitope antibodies and surface plasmon resonance | clinical serum samples | <1 MLD/mL negative in ELS and CIDP patient’s sera |
[167] |
| A | Endopep-MS | clinical serum samples | ~1 MLD/mL | [185] |
| B | Endopeptidase assay and detection with neoepitope antibodies and surface plasmon resonance | clinical serum samples | 0.1–0.01 MLD/mL negative in ELS and GBS patient’s sera |
[166] |
Endopep-MS, endopeptidase assay and mass spectrometry; ELS, Eaton-Lambert syndrome; CIDP, chronic inflammatory demyelinating neuropathy; GBS, Guillain-Barré syndrome. a The equivalence in MLD/mL has been determined according to [186].