Rapid and easy detection of the five most common founder mutations in BRCA1 and BRCA2 genes in the Polish population using CAPS and ACRS-PCR methods

Abstract

In this publication, we present a fast method of diagnosing the most common polymorphisms of BRCA1 and BRCA2 genes in Poland - c.181T>G, pCys61Gly (also known as C61G), c.190T>C, p.Cys64Arg (aka C64R), c.4035delA, p.Glu1346Lysfs (aka 4153delA), c.3700_3704delGTAAA, p.Val1234Glnfs (aka 3819del5), and c.5744C>T, p.Thr1915Met (aka C5972T). Our procedure is based on the use of the cleaved amplified polymorphic sequences (CAPS) and artificially created restriction site (ACRS) PCR techniques. The precise selection of the appropriate primer sequences and restriction enzymes enabled specific cuts of DNA fragments. The final quantity and size of the obtained products depended on the presence or the absence of the mutations. The obtained results are unambiguous and do not have to be confirmed by sequencing. The methods of detection of the c.181T>G, c.190T>C, c.4035delA, c.3700_3704delGTAAA, and c.5744C>T mutations in the BRCA1 and BRCA2 genes described by us do not require the sequencing process, which is more expensive, timeconsuming and associated with numerous errors. The technique developed by us enables the use of simple electrophoresis for accurate detection of the presence or absence of a specific mutation. Our procedure is fast, precise and unambiguous. It is very useful as the first step in the diagnostic of BRCA1/2 constitutional mutations in Polish population in a small clinical laboratory.