Chapleau Richard R, Baldwin James C
Applied Technology and Genomics Center, United States Air Force School of Aerospace Medicine , 711th Human Performance Wing, Air Force Research Laboratory, Wright-Patterson AFB OH .
J Clin Diagn Res. 2015 Aug;9(8):DC24-7. doi: 10.7860/JCDR/2015/13983.6408. Epub 2015 Aug 1.
Optical restriction genome mapping is a technology in which a genome is linearized on a surface and digested with specific restriction enzymes, giving an arrangement of the genome with gaps whose order and size are unique for a given organism. Current applications of this technology include assisting with the correct scaffolding and ordering of genomes in conjunction with whole-genome sequencing, observation of genetic drift and evolution using comparative genomics and epidemiological monitoring of the spread of infections. Here, we investigated the suitability of genome mapping for use in clinical labs as a potential diagnostic tool.
Using whole genome mapping, we investigated the basic performance of the technology for identifying two bacteria of interest for food-safety (Lactobacilli spp. and Enterohemorrhagic Escherichia coli). We further evaluated the performance for identifying multiple organisms from both simple and complex mixtures.
We were able to successfully generate optical restriction maps of four Lactobacillus species as well as a strain of Enterohemorrhagic Escherichia coli from within a mixed solution, each distinguished using a common compatible restriction enzyme. Finally, we demonstrated that optical restriction maps were successfully obtained and the correct organism identified within a clinical matrix.
With additional development, whole genome mapping may be a useful clinical tool for rapid invitro diagnostics.
光学限制性基因组图谱绘制是一种技术,在该技术中,基因组在一个表面上被线性化并用特定的限制性内切酶进行消化,从而得到基因组的一种排列,其中的间隙对于给定生物体而言其顺序和大小是独特的。该技术目前的应用包括结合全基因组测序协助基因组进行正确的支架构建和排序、利用比较基因组学观察遗传漂变和进化以及对感染传播进行流行病学监测。在此,我们研究了基因组图谱绘制作为一种潜在诊断工具在临床实验室中的适用性。
我们使用全基因组图谱绘制技术,研究了该技术用于鉴定两种与食品安全相关的目标细菌(乳酸杆菌属和肠出血性大肠杆菌)的基本性能。我们进一步评估了从简单和复杂混合物中鉴定多种生物体的性能。
我们能够成功地从混合溶液中生成四种乳酸杆菌以及一株肠出血性大肠杆菌的光学限制性图谱,每种都使用一种常见的兼容限制性内切酶进行区分。最后,我们证明在临床样本中成功获得了光学限制性图谱并鉴定出了正确的生物体。
随着进一步发展,全基因组图谱绘制可能成为一种用于快速体外诊断的有用临床工具。