ISSN: 0973-7510

E-ISSN: 2581-690X

Daruosh Momenzadeh1, Sharam Parvin2, Ali Karami3 , Fateme Pourali3, Ali Ahmadi3, Mohammad Heiat3, Hamid Kooshki4, Neda Bayat1 and Fariba Ghasemvand1
1Brain and Spinal injury Research Center, Tehran University of Medical Sciences, Tehran, Iran.
2Chemical Injuries Research Center, Baqiyatallah University of Medical Sciences Tehran, Iran.
3Research Center of Molecular Biology, Baqiyatallah University of Medical Sciences Tehran, Iran.
4Nanobiotechnology Research Center, Baqiyatallah University of Medical Sciences Tehran, Iran.
J Pure Appl Microbiol. 2015;9(Spl. Edn. 1):529-534
© The Author(s). 2015
Received: 08/01/2015 | Accepted: 24/03/2015 | Published: 31/05/2015
Abstract

Bacillus anthracis is a spore-forming bacterium that causes anthrax disease. Its spore resists against physical and chemical agents. Molecular detection of B.anthracis spore requaiers the genomic DNA which is embded in the spores. in this study we developed a rapid detection of spor by Multiplex PCR using rapid physical disruption of spore in a short time without genomic DNA extraction. Genome of non-virulent strain of B.anthracis spores was released by MiniBead Beater-8 system. 3 pairs of primers were used in Multiplex PCR. PCR products were analyzed in agarose gels electrophoresis. Detection limit , specificity and accuracy of the test were determined through various process. This method enables to disrupt spores during 3 minutes and perform detection process with specific primers by Multiplex PCR method. Gel electrophoresis analysis showed 3 fragments of PCR products. digested 1083 bp fragment created 696 bp and 386 bp fragments which  confirmed the test accuracy. Detection limit was 7680 spores/ml or 7 spore/µl. The results showed that the optimized physical disruption method has a proper velosity and there is no need for DNA extraction. Finally this method is capable to detect B.anthracis spores with high sensitivity and specificity.

Keywords

Detection- Bacillus anthracis – Spore – Multiplex PCR

Article Metrics

Article View: 862

Share This Article

© The Author(s) 2015. Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License which permits unrestricted use, sharing, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.