Carret Céline Karine, Horrocks Paul, Konfortov Bernard, Winzeler Elizabeth, Qureshi Matloob, Newbold Chris, Ivens Alasdair
Pathogen Microarrays Group, The Wellcome Trust Sanger Institute, Genome Campus, Hinxton, Cambridge CB10 1SA, UK.
Mol Biochem Parasitol. 2005 Dec;144(2):177-86. doi: 10.1016/j.molbiopara.2005.08.010. Epub 2005 Sep 8.
Microarray-based comparative genomic hybridization (CGH) provides a powerful tool for whole genome analyses and the rapid detection of genomic variation that underlies virulence and disease. In the field of Plasmodium research, many of the parasite genomes that one might wish to study in a high throughput manner are not laboratory clones, but clinical isolates. One of the key limitations to the use of clinical samples in CGH, however, is the miniscule amounts of genomic DNA available. Here we describe the successful application of multiple displacement amplification (MDA), a non-PCR-based amplification method that exhibits clear advantages over all other currently available methods. Using MDA, CGH was performed on a panel of NF54 and IT/FCR3 clones, identifying previously published deletions on chromosomes 2 and 9 as well as polymorphism in genes associated with disease pathology.
基于微阵列的比较基因组杂交(CGH)为全基因组分析以及快速检测作为毒力和疾病基础的基因组变异提供了强大工具。在疟原虫研究领域,许多人们希望以高通量方式研究的寄生虫基因组并非实验室克隆株,而是临床分离株。然而,在CGH中使用临床样本的一个关键限制是可获得的基因组DNA量极少。在此,我们描述了多重置换扩增(MDA)的成功应用,MDA是一种基于非PCR的扩增方法,与目前所有其他可用方法相比具有明显优势。使用MDA,对一组NF54和IT/FCR3克隆株进行了CGH分析,鉴定出先前报道的2号和9号染色体上的缺失以及与疾病病理学相关基因中的多态性。