A PCR-mutagenesis strategy for rapid detection of mutations in codon 634 of the <it>ret</it> proto-oncogene related to MEN 2A.

• Main Authors: Godoy Clara, Perinetti Héctor, Pusiol Eduardo, Roqué María, Mayorga Luis S
• Format: Article
• Language: English
• Published: BMC 2002-05-01
• Series: BMC Medical Genetics
• Online Access: http://www.biomedcentral.com/1471-2350/3/4

<p>Abstract</p> <p>Background</p> <p>Multiple endocrine neoplasias type 2A (MEN 2A) is a dominantly inherited cancer syndrome. Missence mutations in the codon encoding cysteine 634 of the <it>ret</it> proto-oncogene have been found in 85% of the MEN 2A families. The main tumour type always present in MEN 2A is medullar thyroid carcinoma (MTC). Only 25% of all MTC are hereditary, and generally they are identified by a careful family history. However, some familial MTCs are not easily detected by this means and underdiagnosis of MEN 2A is suspected.</p> <p>Methods</p> <p>DNA samples from MEN 2A patients were amplified by PCR. The products were incubated with the restriction enzyme Bst ApI or Bgl I.</p> <p>The samples were loaded in non-denaturing 10% Polyacrilamyde Gel and run at 120 volts for 40 min. The gels were stained with 10 μg/ml ethidium bromide, and the bands were visualized under a UV lamp.</p> <p>Results</p> <p>We developed a PCR-mutagenic method to check the integrity of the three bases of the cysteine 634 codon.</p> <p>Conclusion</p> <p>The method can be used to detect inherited mutations in MTC patients without a clear family history. The method is relatively simple to use as a routine test in these patients to decrease the underdiagnosis of MEN 2A. In addition, the assay can be used to screen affected families with any mutation in cysteine 634.</p>