Abstract
Ricin and abrin are two of the most potent plant toxins known and may be easily obtained in high yield from the seeds using rather simple technology. As a result, both toxins are potent and available toxins for criminal or terrorist acts. However, as the production of highly purified ricin or abrin requires sophisticated equipment and knowledge, it may be more likely that crude extracts would be used by non-governmental perpetrators. Remaining plant-specific nucleic acids in these extracts allow the application of a real-time PCR (qPCR) assay for the detection and identification of abrin or ricin genomic material. Therefore, we have developed a duplex real-time PCR assays for simultaneous detection of ricin and abrin DNA based on the OmniMix HS bead PCR reagent mixture. Novel primers and hybridization probes were designed for detection on a SmartCycler instrument by using 5′-nuclease technology. The assay was thoroughly optimized and validated in terms of analytical sensitivity. Evaluation of the assay sensitivity by probit analysis demonstrated a 95% probability of detection at 3 genomes per reaction for ricin DNA and 1.2 genomes per reaction for abrin DNA. The suitability of the assays was exemplified by detection of ricin and abrin contaminations in a food matrix.
Highlights
Accidental and intentional intoxications from lectins in humans and animals have been known for centuries [1]
The main causative agents hereby belong to the family of Euphorbiaceae (Ricinus communis) and Fabaceae (Abrus precatorius) [2,3]
Oligonucleotide primers were designed manually based on an alignment of Ricinus communis, Abrus precatorius and other lectin-producing plant genome information
Summary
Accidental and intentional intoxications from lectins in humans and animals have been known for centuries [1]. Two of the most potent toxins known, are produced in the seeds of the jequirity plant (Abrus precatorius) (Figure 1 a,b) and the castor plant (Ricinus communis) (Figure 1 c,d) respectively. Both plants are perennial in tropical and subtropical regions of the world. Abrin and ricin exert toxicity through inhibition of protein synthesis, which results in cell death. Both toxins belong to the large family of ribosome-inactivating proteins and contain two disulfide-linked heterodimeric chains (A and B) with similar molecular mass (32 and 34 kDa). As part of an effort to provide rapid and reliable methods for the detection of various biological agents including plant toxins, a qPCR assay for simultaneous detection of ricin and abrin nucleic acid was developed
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