Alternative Test Methods: Validation and Regulatory Status
- Acute Aquatic Toxicity
- Acute Mammalian Oral Toxicity
- Acute Mammalian Inhalation Toxicity
- Acute Mammalian Skin Toxicity
- Biologics (non-vaccine)
- Carcinogenicity
- Chronic/Repeated Dose Toxicity
| Conventional test method | Alternative Test Method | Infor- mation Last Updated | ||||
|---|---|---|---|---|---|---|
| Name & Description | Validation Status | Regulatory Status | Effect or Potential Effect on Animal Use | |||
| Biologics (non-vaccine) | ||||||
Mouse LD50 Assay for Botulinum Toxin Potency Testing |
Endopeptidase Assay—SNAP 25 Description and references |
UK: Validated to verify potency by the National Institute for Biological Standards and Control (NIBSC) (1999) US: ICCVAM workshop panel concluded that it is possible, in principle, to use an endopeptidase assay for estimating BoNT concentration in a pharmaceutical preparation. However, they cannot currently be used to replace animals for BoNT potency testing or for detecting BoNT in environmental or biological samples. Future activities were recommended (2008). |
EU: Listed in the European Pharmacopoeia UK: Routinely used by the NIBSC US: None |
Replacement | April 2009 | |
| Endopeptidase-MS assay Description and references |
US: ICCVAM workshop panel concluded that it is possible, in principle, to use an endopeptidase assay for estimating BoNT concentration in a pharmaceutical preparation. However, they cannot currently be used to replace animals for BoNT potency testing or for detecting BoNT in environmental or biological samples. Future activities were recommended. (2008) |
None |
Replacement | April 2009 | ||
| Mouse Hind-Limb Assay (Digital Abduction Score (DAS) Assay) Description and references |
US: ICCVAM workshop panel concluded that the DAS assay is applicable to multiple serotypes and products, and has potential as a potency assay, if used relative to a reference standard (a standardized, calibrated BoNT preparation against which to establish “relative” potency values). Recommended that further validation studies be conducted (2008). |
EU: Listed in the European Pharmacopoeia (2005) US: None |
Refinement (Does not rely on death as an endpoint) |
April 2009 | ||
| Mouse Abdominal Ptosis Assay or Flaccid Paralysis (Non-lethal Mouse Model for Botulinum Toxin Potency Testing) Description and references |
UK: Validated at NIBSC for potency testing of BoNT products US: ICCVAM workshop panel concluded that the Abdominal Ptosis Assay |
EU: Listed in the European Pharmacopoeia UK: Used by NIBSC when in vitro test is inconclusive US: None |
Refinement (Does not rely on death as an endpoint) |
April 2009 | ||
| Non-Lethal Clinical Endpoint in the Mouse LD50 Assay Protocol
Description and references |
Canada: Validated by Health Canada US: ICCVAM workshop panel recommended that an international validation study using the same non-lethal endpoint be conducted (2008). |
Canada: Accepted for use |
Refinement (Does not rely on death as an endpoint) |
April 2009 | ||
Mouse Phrenic Nerve (MPN) Assay |
US: ICCVAM workshop panel recommended that further validation studies be conducted (2008). Germany: Validated by the Medical School of Hanover |
EU: Listed in the European Pharmacopoeia (2005) |
Reduction Refinement |
April 2009 | ||
Rat Intercostal Neuromuscular Junction (NMJ) Assay |
US: ICCVAM workshop panel recommended that further validation studies be conducted (2008) UK: Currently undergoing validation |
None |
Reduction Refinement |
April 2009 | ||
In Vitro Cell-Based Assay |
US: ICCVAM workshop panel found that no cell-based method could currently be used to reduce or replace animals for BoNT potency testing or for detecting botulinum toxin in environmental or biological samples. In their current state of development, cell-based methods are not sufficiently sensitive. Which cell types and/or cell lines are the most appropriate is also not known (2008). |
None |
Replacement | April 2009 | ||
List of Abbreviations
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