development of pcr-based method for rapid detection of abrin gene

نویسندگان

morteza hamedi behnaq molecular biology research center, baqiyatallah university of medical sciences, tehran, iran

ali karami molecular biology research center, baqiyatallah university of medical sciences, tehran, iran e-mail: [email protected]

ali choopani 2. applied biotechnology research center, baqiyatallah university of medical sciences, teh-ran, iran 3. department of biology, payam noor university, tehran, iran.

چکیده

abrin known as ribosome inactivating protein’s (rip) it is a high cytotoxic plant protein. high lethality and low cost and easy access to plant and seeds lead to this toxin used in crimes and terrorist acts. because obtaining purified toxins requires advanced laboratory equipment and complex procedures, so, it appears that the perpetrators of such crimes use crude extracts. as a result, remaining of specific toxin gene in these extracts take the advantage of using pcr assay to identify abrin gene, that refers to exist of its toxin. we use new rapid molecular method for abrin gene detection by pcr. we designed specific primers for detection of abrin toxins gene by pcr. dna was extracted by cetyl trimethylammonium bromide-polyvinylpyrrolidone (ctab-pvp) method from rosary pie samples and pcr protocol was performed using primers for toxin gene amplification. then we analyzed assay’s sensitivity by serial dilution method. the results of this study revealed that designed and selected primers sequence for toxin’s gene is specific. the desired product size was obtained and sequencing of pcr products was performed that show 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 is sensitive, and low-cost to detect this toxins gene in cases of suspected to bioterrorism event.

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عنوان ژورنال:
applied biotechnology reports

جلد ۳، شماره ۳، صفحات ۴۷۳-۰

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