Awad Mohamed, Ouda Osama, El-Refy Ali, El-Feky Fawzy A, Mosa Kareem A, Helmy Mohamed
Department of Biotechnology, Faculty of Agriculture, Al-Azhar University, Cairo 11651, Egypt.
Department of Information Technology, Faculty of Computer and Information Sciences, Mansoura University, Mansoura 35516, Egypt.
Adv Bioinformatics. 2015;2015:303605. doi: 10.1155/2015/303605. Epub 2015 Dec 31.
Sequencing and restriction analysis of genes like 16S rRNA and HSP60 are intensively used for molecular identification in the microbial communities. With aid of the rapid progress in bioinformatics, genome sequencing became the method of choice for bacterial identification. However, the genome sequencing technology is still out of reach in the developing countries. In this paper, we propose FN-Identify, a sequencing-free method for bacterial identification. FN-Identify exploits the gene sequences data available in GenBank and other databases and the two algorithms that we developed, CreateScheme and GeneIdentify, to create a restriction enzyme-based identification scheme. FN-Identify was tested using three different and diverse bacterial populations (members of Lactobacillus, Pseudomonas, and Mycobacterium groups) in an in silico analysis using restriction enzymes and sequences of 16S rRNA gene. The analysis of the restriction maps of the members of three groups using the fragment numbers information only or along with fragments sizes successfully identified all of the members of the three groups using a minimum of four and maximum of eight restriction enzymes. Our results demonstrate the utility and accuracy of FN-Identify method and its two algorithms as an alternative method that uses the standard microbiology laboratories techniques when the genome sequencing is not available.
16S rRNA和HSP60等基因的测序和限制性分析在微生物群落的分子鉴定中被广泛应用。随着生物信息学的迅速发展,基因组测序成为细菌鉴定的首选方法。然而,基因组测序技术在发展中国家仍然难以企及。在本文中,我们提出了FN-Identify,一种无需测序的细菌鉴定方法。FN-Identify利用GenBank和其他数据库中可用的基因序列数据以及我们开发的两种算法CreateScheme和GeneIdentify,创建基于限制性内切酶的鉴定方案。在使用限制性内切酶和16S rRNA基因序列的计算机模拟分析中,我们使用三种不同且多样的细菌群体(乳酸杆菌属、假单胞菌属和分枝杆菌属的成员)对FN-Identify进行了测试。仅使用片段数量信息或结合片段大小对三组成员的限制性图谱进行分析,成功地使用最少四种和最多八种限制性内切酶鉴定了三组中的所有成员。我们的结果证明了FN-Identify方法及其两种算法作为一种替代方法的实用性和准确性,当无法进行基因组测序时,该方法可使用标准微生物实验室技术。