Multiplex ligation-dependent probe amplification using a completely synthetic probe set

被引:68
作者
Stern, RF [1 ]
Roberts, RG [1 ]
Mann, K [1 ]
Yau, SC [1 ]
Berg, J [1 ]
Ogilvie, CM [1 ]
机构
[1] Guys & St Thomas Hosp, Sch Med, Dept Med & Mol Genet, Guys Hosp, London SE1 9RT, England
关键词
D O I
10.2144/04373ST04
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The recent development of multiplex ligation-dependent probe amplification (MLPA) has provided an efficient and reliable assay for dosage screening of multiple loci in a single reaction. However; a drawback to this method is the time-consuming process of generating a probe set by cloning in single-stranded bacteriophage vectors. We have developed a synthetic probe set to screen for deletions in a region spanning 18.5 Mb within chromosome 3q. In a pilot study, we tested 15 synthetic probes on 4 control samples and on 2 patients previously found to possess a heteroygous deletion in the region 3q26-q28. These synthetic probes detected deletions at all previously known deleted loci. Furthermore, using synthetic probes, the variability of results within samples was similar to that reported for commercially available M13-derived probes. Our results demonstrate that this novel approach to MLPA provides a generic solution to the difficulties of probe development by cloning; such synthetically generated probes may be used to screen a large number of loci in a single reaction. We conclude that the use of synthetic probes for MLPA is a rapid, robust, and efficient alternative for research (and potentially diagnostic) deletion and duplication screening of multiple genomic loci.
引用
收藏
页码:399 / 405
页数:7
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