Botulism is a severe and potentially lethal paralytic disease caused by several
botulinum neurotoxin-producing Clostridia spp. In China, the majority of the cases caused by
botulism were from less-developed rural areas. Here, we designed specific substrate
peptides and reconfigured
gold nanoparticle-based lateral flow test strip (LFTS) to develop an
endopeptidase-based lateral flow assay for the diagnosis of
botulism. We performed this lateral flow assay on
botulinum neurotoxin-spiked human serum samples. The as-prepared LFTS had excellent performance in the detection of
botulinum neurotoxin using only 1 μL of simulated serum, and its sensitivity and specificity were comparable to that of mouse lethality assay. Moreover, the assay takes only half a day and does not require highly trained laboratory staff, specialized facility, or equipment. Finally, our LFTS can be potentially extended to other serotypes of BoNTs by designing specific substrate
peptides against the different types of BoNTs. Overall, we demonstrate a strategy by which LFTS and
endopeptidase activity assays can be integrated to achieve facile and economic diagnosis of
botulism in resource-limited settings.