Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Filters applied. Clear all
. 2012 Sep;4(9):633-42.
doi: 10.3390/toxins4090633. Epub 2012 Aug 31.

Simultaneous Detection of Ricin and Abrin DNA by Real-Time PCR (qPCR)

Affiliations
Free PMC article

Simultaneous Detection of Ricin and Abrin DNA by Real-Time PCR (qPCR)

Eva Felder et al. Toxins (Basel). .
Free PMC article

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.

Keywords: abrin; quantitative real-time PCR; ricin; simultaneous detection.

Figures

Figure 1
Figure 1
(a) Abrus precatorius plant and (b) jequirity seeds. (c) Ricinus communis plant and (d) castor seeds.
Figure 2
Figure 2
Primers and Probes used in this study; the oligonucleotide binding sites within the genome of different lectin-producing plants are shown. The scale refers to the position on the genome of Ricinus communis (XM_002532143.1).
Figure 3
Figure 3
Limiting dilution series of 107–100 copies DNA per reaction; NTC: no template control (a) A. precatorius DNA and (b) R. communis (cultivar Carmencita pink) DNA. For further use as positive control for verification of assay performance quantified PCR products were adjusted to result in final crossing point values of 30 and dried down in a SpeedVac vacuum centrifuge for long term storage.
Figure 4
Figure 4
Probit analysis performed with the duplex qPCR protocol: Probability of achieving a positive test result (y-axis), depending on the DNA copy input number per PCR of (a) A. precatorius and (b) R. communis (cultivar Carmencita pink) DNA (x-axis). Upper and lower curves represent 95% confidence limits of the model.

Similar articles

See all similar articles

Cited by 9 articles

See all "Cited by" articles

References

    1. Flexner S. The histological changes produced by Ricin and Abrin intoxications. J. Exp. Med. 1897;2:197–216. doi: 10.1084/jem.2.2.197. - DOI - PMC - PubMed
    1. Carter S. Euphorbia. In: Eggli U., Albers F., editors. Dicotyledons. Springer; Berlin, Germany: 2001. p. 102.
    1. Dickers K.J., Bradberry S.M., Rice P., Griffiths G.D., Vale J.A. Abrin poisoning. Toxicol. Rev. 2003;22:137–142. doi: 10.2165/00139709-200322030-00002. - DOI - PubMed
    1. Nass L.L., Pereira P.A.A., Ellis D. Biofuels in Brazil: An overview. Crop Sci. 2007;47:2228–2237. doi: 10.2135/cropsci2007.03.0166. - DOI
    1. Worbs S., Kohler K., Pauly D., Avondet M.A., Schaer M., Dorner M.B., Dorner B.G. Ricinus communis intoxications in human and veterinary medicine-a summary of real cases. Toxins. 2011;3:1332–1372. doi: 10.3390/toxins3101332. - DOI - PMC - PubMed
Feedback