Why Use MLPA®?
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).

Simultaneous detection

of copy numbers, methylation and selected point mutations.
Low input


Requires only 50 ng of DNA.
Time-efficient


Results available within 24 hours.
Short hands-on time

MLPA is performed in 5 simple steps.
Cost-effective


One MLPA reaction costs EUR 12/USD 15.

Brochures
Our Support Portal contains several brochures that provide an easy overview of the MLPA technique and some of its major applications. You can find these brochures in the downloads section (link opens in a new window).

References
  1. Schouten JP et al. (2002) Relative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplification Nucleic Acids Res 30, e57.
  2. Aretz, S. et al. (2007). High proportion of large genomic deletions and a genotype phenotype update in 80 unrelated families with juvenile polyposis syndrome J Med Genet. 44, 702-709.
  3. Redeker, E.J., et al. (2008). Multiplex ligation-dependent probe amplification (MLPA) enhances the molecular diagnosis of aniridia and related disorders Mol Vis 14, 836-840.
  4. Kanno, J., et al. (2007). Genomic deletion within GLDC is a major cause of non-ketotic hyperglycinaemia J Med Genet 44, 3.
  5. Aldred, M.A., et al. (2006). BMPR2 gene rearrangements account for a significant proportion of mutations in familial and idiopathic pulmonary arterial hypertension Hum Mutat 2, 212-213.
  6. Kluwe, L., et al. (2005). Screening for large mutations of the NF2 gene Genes Chromosomes Cancer 42, 384-391.
  7. Michils, G., et al. (2005). Large deletions of the APC gene in 15% of mutation-negative patients with classical polyposis (FAP): a Belgian study Hum Mutat 2, 125-34.
  8. Taylor, C.F., et al. (2003). Genomic deletions in MSH2 or MLH1 are a frequent cause of hereditary non-polyposis colorectal cancer: identification of novel and recurrent deletions by MLPA Hum Mutat 6, 428-33.
  9. Depienne, C., et al. (2007). Exon deletions of SPG4 are a frequent cause of hereditary spastic paraplegia J Med Genet 44, 281-284.
  10. Beetz, C., et al. (2006). High frequency of partial SPAST deletions in autosomal dominant hereditary spastic paraplegia Neurology 67, 1926-1930.
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