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Simultaneous detection of ricin and abrin DNA by real-time PCR (qPCR).

Felder E, Mossbrugger I, Lange M, Wölfel R - Toxins (Basel) (2012)

Bottom Line: 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.Novel primers and hybridization probes were designed for detection on a SmartCycler instrument by using 5'-nuclease technology.

View Article: PubMed Central - PubMed

Affiliation: Department for Medical Bio Reconnaissance and Verification, Bundeswehr Institute of Microbiology, Neuherbergstrasse 11, Munich 80937, Germany. evafelder@bundeswehr.org

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.

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(a) Abrus precatorius plant and (b) jequirity seeds. (c) Ricinus communis plant and (d) castor seeds.
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toxins-04-00633-f001: (a) Abrus precatorius plant and (b) jequirity seeds. (c) Ricinus communis plant and (d) castor seeds.

Mentions: 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]. Abrin and ricin, 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. Although the lethal toxicity of ricin and abrin is about 1000-fold less than that of botulinum toxin, accessibility, and ease of production in massive quantities make especially ricin a potential biological agent for terrorism. Today about 1.4 million metric tons of castor seeds are harvested annually for castor oil production [4]. Ricin as a water-soluble protein is not extracted into the castor oil, therefore industrial grade castor oil is found to be safe [5]. However, the press cake that remains after pressing and solvent extraction contains virtually all of the ricin present in the seeds (1%–5% ricin w/w), making at least ricin an easily available, inexpensive and easy to prepare toxin. The potential threat of abrin and ricin is considered significant and accordingly both toxins are classified as category B Select Agents by US Health and Human Services.


Simultaneous detection of ricin and abrin DNA by real-time PCR (qPCR).

Felder E, Mossbrugger I, Lange M, Wölfel R - Toxins (Basel) (2012)

(a) Abrus precatorius plant and (b) jequirity seeds. (c) Ricinus communis plant and (d) castor seeds.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3475220&req=5

toxins-04-00633-f001: (a) Abrus precatorius plant and (b) jequirity seeds. (c) Ricinus communis plant and (d) castor seeds.
Mentions: 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]. Abrin and ricin, 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. Although the lethal toxicity of ricin and abrin is about 1000-fold less than that of botulinum toxin, accessibility, and ease of production in massive quantities make especially ricin a potential biological agent for terrorism. Today about 1.4 million metric tons of castor seeds are harvested annually for castor oil production [4]. Ricin as a water-soluble protein is not extracted into the castor oil, therefore industrial grade castor oil is found to be safe [5]. However, the press cake that remains after pressing and solvent extraction contains virtually all of the ricin present in the seeds (1%–5% ricin w/w), making at least ricin an easily available, inexpensive and easy to prepare toxin. The potential threat of abrin and ricin is considered significant and accordingly both toxins are classified as category B Select Agents by US Health and Human Services.

Bottom Line: 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.Novel primers and hybridization probes were designed for detection on a SmartCycler instrument by using 5'-nuclease technology.

View Article: PubMed Central - PubMed

Affiliation: Department for Medical Bio Reconnaissance and Verification, Bundeswehr Institute of Microbiology, Neuherbergstrasse 11, Munich 80937, Germany. evafelder@bundeswehr.org

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.

Show MeSH