Rapid RT-PCR-based protein truncation test in the screening for 5′ located mutations of the APC gene

Althoughin vitroprotein synthesis is a rapid method to screen for translational stops in the adenomatous polyposis coli (APC) gene, truncating mutations at the 5′ most end are at risk of being overseen due to their small size. The authors describe a reverse transcriptase-polymerase chain reaction (RT-PCR)-based protein truncation test specifically designed for detecting truncated polypeptide chains of less than 10 kDa. Using this detection system, three novel germline mutations in familial adenomatous polyposis (FAP) patients were identified, i.e. a Gly101Ter non-sense mutation in exon 3, an exon 4 splice acceptor mutation and a 555delC deletion in exon 5. Moreover, a patient manifesting congenital hypertrophy of the retinal pigmented epithelium (CHRPE) was detected with an Arg232Ter mutation in exon 6. This is, to the authors» knowledge, the fourth exception to the rule that FAP patients manifesting CHRPE harbour genetic alterations downstream from APC exon 9. Hence, an alternative hotspot for non-sense mutations associated with CHRPE appears to encompass the codons 215, 216 and 232. Patients reported in this study, exhibited relatively mild clinical symptoms with respect to the age of onset of malignancy (>50 years of age) and the number of polyps (70–100 adenomas). However, manifestation of severe duodenal adenomatosis was independent of the attenuated colorectal FAP phenotype.