Development of PCR-Based Method for Rapid Detection of Abrin Gene

Document Type: Original Article

Authors

1 Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

2 Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

3 Department of Biology, Payam Noor University, Isfahan, Iran

Abstract

Abrin, known as a ribosome inactivating protein (RIP), is a high cytotoxic plant protein. The high lethality, low cost, and easy access to this plant and its seeds have led to this toxin to be used in crimes and terrorist acts. Since, obtaining purified toxins requires advanced laboratory equipment and complex procedures, it seems that the perpetrators of such crimes use crude extracts. As a result, it was hypothesized that remaining the specific toxin genes in these extracts can provide the advantage of using PCR assay to identify abrin gene which refers to the existence of its toxin. We used a new rapid molecular method for the detection of the abrin gene by PCR. In this regard, specific primers were designed and the required DNA was extracted from Rosary pie samples using cetyltrimethylammonium bromide-polyvinylpyrrolidone (CTAB-PVP) method and PCR protocol was performed using specific primers. Then, assay’s sensitivity was analyzed using serial dilution method. The results of this study revealed that designed and selected primers sequence for toxin’s gene function as specific. The desired product size was obtained and sequencing of PCR products showed up to 90% similarity with known sequence for each molecule. According to these results, the developed rapid molecular method for detection of abrin toxin gene can be considered as a sensitive and low-cost detection method for this toxin gene in cases of suspection to bioterrorism event.

Keywords