J Korean Med Sci.  2009 Feb;24(1):77-83. 10.3346/jkms.2009.24.1.77.

Improved Detection of Germline Mutations in Korean VHL Patients by Multiple Ligation-dependent Probe Amplification Analysis

• Cho HJ
• Ki CS
• Kim JW

• KMID: 1794410
• DOI: http://doi.org/10.3346/jkms.2009.24.1.77

Abstract

von Hippel-Lindau (VHL) disease is an autosomal dominant inherited tumor syndrome characterized by the development of tumors in the eye, brain, spinal cord, inner ear, adrenal gland, pancreas, kidney, and epididymis, associated with germline mutations in the VHL gene. We used sequentially sequencing method and multiple ligation-dependent probe amplification (MLPA) analysis and detected germline mutations in the VHL in 15/15 (100%) of VHL patients fulfilling the clinical criteria. Of the 15 distinct mutations detected, large deletions were detected in 5/15 (33.3%) patients, including 4/15 (26.7%) partial deletions and 1/15 (6.6%) deletion of the entire VHL gene by MLPA and the remainder were point mutations detected by sequencing method, of which five mutations were novel. Using MLPA analysis, we detected large deletions including both partial deletions and complete gene deletion, which has not been reported in Korean VHL patients. In conclusion, sequential application of sequencing method and MLPA analysis might make possible to identify germline mutations in most patients with VHL.

Keyword

VHL; MLPA; Gene Deletion; Korean

MeSH Terms

Adolescent
Adult
Asian Continental Ancestry Group/*genetics
DNA Mutational Analysis/*methods
Female
Gene Deletion
Genetic Predisposition to Disease
*Germ-Line Mutation
Humans
Korea
Male
Middle Aged
*Nucleic Acid Amplification Techniques
Phenotype
Polymorphism, Single Nucleotide
Sequence Analysis
Young Adult
von Hippel-Lindau Disease/diagnosis/*genetics

Figure

• 

  Fig. 1 Multiplex ligation-dependent probe amplification (MLPA) analysis of VHL exons 1-3. Signal intensity reveals only a single copy of VHL exon 2 and 3 in a patient with VHL due to deletion of VHL exon 2 and 3 (A) and only a single copy of each VHL exon in a patient with VHL due to deletion of whole VHL gene deletion (B) when compared to a healthy control (C).

• 

  Fig. 2 DNA dosage profiles estimated by MLPA analysis. Ratios on each DNA dosage are plotted relative to single DNA samples from five unrelated healthy controls. Cut-off levels for loss of relative copy numbers were set at 0.7. A patient with VHL (patient 6) due to deletion of VHL exon 2 and 3 (A) and a patient with VHL (patient 10) due to deletion of whole VHL gene deletion (B). Ex×1, Exon 1; Ex×2, Exon 2; Ex×3, Exon 3.

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