Telomerase reverse transcriptase promoter (TERTp) mutations are frequently targeted
tumor markers, however, they reside in regions with high GC content, which poses challenges
when examined with simple molecular techniques or even with next-generation sequencing
(NGS). In bladder cancer (BC), TERTp mutations are particularly frequent, however,
none of the available tools have demonstrated efficacy in detecting TERTp mutations
via a simple noninvasive technique. Therefore, we developed a novel PCR-based method
for the detection of the two most common TERTp mutations and demonstrated its use
for the analysis of BC samples. The developed SHARD-PCR TERTp mutation detection technique
requires PCR and restriction digestion steps that are easily implementable even in
less well-equipped laboratories. Cell lines with known mutational status were utilized
for method development. Matching urine and tumor tissue samples from BC patients were
analyzed, and the results were validated by next-generation sequencing. Analysis of
eighteen urine and corresponding tumor tissue samples by SHARD-PCR revealed perfect
matches in sample pairs, which paralleled the corresponding NGS results: fourteen
samples exhibited mutations at the -124 position, two samples showed mutations at
the -146 position, and no mutations were detected in two samples. Our study serves
as a proof-of-concept and is limited by its small sample size, nonetheless, it demonstrates
that SHARD-PCR is a simple, economic and highly reliable method for detecting TERTp
mutations, which are common in different cancer types. For bladder cancer, SHARD-PCR
can be performed with the use of noninvasive samples and could replace or complement
currently used techniques.
