MRC-Holland
MLPA®: An Introduction
MLPA (Multiplex Ligation-dependent Probe Amplification) is a multiplex PCR method detecting abnormal copy numbers of up to 60 different genomic DNA sequences, which is able to distinguish sequences differing in only one nucleotide (1). The MLPA technique is easy to use and can be performed in many laboratories, as it only requires a thermocycler and capillary electrophoresis equipment. Up to 96 samples can be handled simultaneously, with results being available within 24 hours. Although for most hereditary conditions, (partial) gene deletions or duplications account for less than 10% of all disease-causing mutations, for many other disorders this is 10 to 30% (2-8) or even higher still (9, 10). The inclusion of MLPA in clinical settings can therefore significantly increase the detection rate of many genetic disorders.
SALSA MLPA kits for a rapidly growing number of applications are available from MRC-Holland (Amsterdam, the Netherlands). A list of available SALSA MLPA kits can be found here.
Advantages of MLPA
Using MLPA for copy number detection offers many advantages over other techniques. First of all, methods which were primarily developed for detecting point mutations, such as sequencing and DHPLC, generally fail to detect copy numbers changes. Southern blot analysis, on the other hand, will reveal many aberrations but will not always detect small deletions and is not ideal as a routine technique. Although well-characterised deletions and amplifications can be detected by PCR, the exact breakpoint site of most deletions is unknown. Furthermore, when comparing MLPA to FISH, MLPA not only has the advantage of being a multiplex technique, but also one in which very small (50-70 nt) sequences are targeted, enabling MLPA to identify the frequent, single gene aberrations which are too small to be detected by FISH. Moreover, MLPA can be used on purified DNA. Finally, as compared to array CGH, MLPA is a low cost and technically uncomplicated method. Although MLPA is not suitable for genome-wide research screening, it is a good alternative to array-based techniques for many routine applications. The over 300 probe sets now commercially available are dedicated to applications ranging from the relatively common (Duchenne, DiGeorge syndrome, SMA) to the very rare (hereditary pancreatitis, Antithrombin deficiency, Birt-Hogg-Dube syndrome).
MLPA®是DNA拷贝数定量的金标准，用于研究遗传性疾病和肿瘤。 MLPA也可用于研究DNA序列的甲基化状态。 MRPA的发明者和制造商MRC-Holland提供400种不同的MLPA面板。热门应用包括：
对癌症的易感性
神经肌肉疾病
智力残疾
实体瘤
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