Document Detail

Array-MLPA: comprehensive detection of deletions and duplications and its application to DMD patients.
MedLine Citation:
PMID:  17854090     Owner:  NLM     Status:  MEDLINE    
Multiplex ligation-dependent probe amplification (MLPA) is widely used to screen genes of interest for deletions and duplications. Since MLPA is usually based on size-separation of the amplification products, the maximum number of target sequences that can be screened in parallel is usually limited to approximately 40. We report the design of a robust array-based MLPA format that uses amplification products of essentially uniform size (100-120 bp) and distinguishes between them by virtue of incorporated tag sequences. We were thus able to increase probe complexity to 124, with very uniform product yields and signals that have a low coefficient of variance. The assay designed was used to screen the largest set studied so far (249 patients) of unrelated Duchenne muscular dystrophy (DMD) cases from the Chinese population. In a blind study we correctly assigned 98% of the genotypes and detected rearrangements in 181 cases (73%); i.e., 163 deletions (65%), 13 duplications (5%), and five complex rearrangements (2%). Although this value is significantly higher for Chinese patients than previously reported, it is similar to that found for other populations. The location of the rearrangements (76% in the major deletion hotspot) is also in agreement with other findings. The 96-well flow-through microarray system used in this research provides high-throughput and speed; hybridization can be completed in 5 to 30 minutes. Since array processing and data analysis are fully automated, array-MLPA should be easy to implement in a standard diagnostic laboratory. The universal array can be used to analyze any tag-modified MLPA probe set.
Fanyi Zeng; Zhao-Rui Ren; Shang-Zhi Huang; Margot Kalf; Monique Mommersteeg; Maarten Smit; Stefan White; Chun-Lian Jin; Miao Xu; Da-Wen Zhou; Jing-Bin Yan; Mei-Jue Chen; Rinie van Beuningen; Shu-Zhen Huang; Johan den Dunnen; Yi-Tao Zeng; Ying Wu
Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Human mutation     Volume:  29     ISSN:  1098-1004     ISO Abbreviation:  Hum. Mutat.     Publication Date:  2008 Jan 
Date Detail:
Created Date:  2007-12-27     Completed Date:  2008-02-06     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  9215429     Medline TA:  Hum Mutat     Country:  United States    
Other Details:
Languages:  eng     Pagination:  190-7     Citation Subset:  IM    
Copyright Information:
(c) 2007 Wiley-Liss, Inc.
Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai Jiao Tong University School of Medicine, PR China.
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MeSH Terms
Gene Deletion*
Gene Duplication*
Genetic Testing / methods*
Muscular Dystrophy, Duchenne / diagnosis*,  genetics*
Nucleic Acid Amplification Techniques / methods*

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