Detection and Measurement of Botulinum Neurotoxin

Tests to detect and measure botulinum neurotoxin (BoNT) are required by multiple federal agencies for a variety of purposes, such as measuring toxin in drug formulations or detecting toxin in possibly contaminated food or wildlife. Currently, the standard test for these endpoints is a mouse lethality assay that can use large numbers of animals. NICEATM supports efforts to develop, validate, and implement alternative approaches for tests used to detect the presence of BoNT and measure potency of BoNT preparations.

Optimization and Validation of an In Vitro Botulinum Neurotoxin Assay

NICEATM supported the optimization and validation of ELISAs that replace animal-based methods for diagnosing suspected avian botulism samples. Methods were developed for determining the presence or absence of BoNT serotypes C, D, and E in field-collected samples from a wide range of bird species. These methods may be used in the future to support testing requirements for the U.S. Geological Survey’s National Wildlife Health Center.

Request for Data and Information on Technologies

In an 2018 Federal Register notice, NICEATM requested available data and information on approaches and/or technologies currently used to detect and measure BoNT. Submitted information will be used to assess the state of the science and determine technical needs for non-animal test methods that are used to detect the presence of BoNT and measure potency of BoNT preparations.

Federal Register notice announcing data request (June 13, 2018) – View as a webpage

Respondents should provide information on any activities relevant to the development or validation of alternatives to in vivo test methods currently used by federal agencies for regulatory and other decision contexts.  NICEATM also requests available data from in vivo BoNT tests used for similar applications as the proposed alternative, such as distinguishing between BoNT serotypes in biological matrix samples or measuring the potency of therapeutic BoNT preparations. View guidelines for submission of data to NICEATM

Respondents to this request for information should include their name, affiliation (if applicable), mailing address, telephone, email, and sponsoring organization (if any) with their communications.   Responses to this notice will be posted on this page, therefore no proprietary, classified, confidential, or sensitive information should be included in responses. Persons submitting responses will be identified on the web page by name and affiliation or sponsoring organization, if applicable. View NTP guidelines for public comments

Responses Received to the June 2018 Data Request

Workshop on Alternatives to the Mouse LD50 Assay for Botulinum Toxin Testing (2006)

In collaboration with international partners, NICEATM and ICCVAM sponsored a workshop to review methods proposed to replace, reduce, and refine the use of mice for BoNT testing. The workshop identified priorities for research, development, and validation efforts. These efforts are needed to advance the use of alternative methods. A report of the workshop was published in 2008. 

A cell-based assay method discussed at the workshop was later accepted by the FDA (2011) for stability and potency testing of BoNT type A products.