TERTmonitor Efficacy and Performance in Detecting Mutations by Droplet Digital PCR

Genes (Basel). 2024 Oct 31;15(11):1424. doi: 10.3390/genes15111424.

Abstract

Background: The screening of TERT promoter (TERTp) mutations is essential in cancer research and diagnostics, due to its prevalence in tumours associated with low self-renewal rates. TERTmonitor is a diagnosis kit primarily designed for real-time qPCR qualitative detection of -124C>T and -146C>T TERTp mutations, which are highly prevalent in several malignancies, particularly in bladder carcinoma.

Objective: This study aims to investigate TERTmonitor performance in droplet digital PCR (ddPCR) in urine samples from bladder cancer patients.

Methods: A total of 45 urine samples were examined by real-time qPCR and ddPCR techniques, and their performances were compared.

Results: TERTmonitor had similar performance in both real-time qPCR and ddPCR platforms. Specifically, the methods exhibited a concordance rate of 95.45% and 90% for -124C>T and -146C>T mutations, respectively. Importantly, an enhanced sensitivity in certain scenarios was exhibited by ddPCR when compared to real-time qPCR, detecting mutations that the latter failed to identify in approximately 4.55% and 10% of the samples for -124C>T and -146C>T mutations, respectively. This enhanced sensitivity of ddPCR was particularly evident in samples with low-frequency mutations.

Conclusions: The findings highlight the usefulness of TERTmonitor for cancer surveillance either in real-time qPCR or ddPCR platforms.

Keywords: TERTmonitor; ddPCR; mutations; real-time qPCR; telomerase promoter.

MeSH terms

  • Aged
  • Female
  • Humans
  • Male
  • Middle Aged
  • Mutation*
  • Promoter Regions, Genetic
  • Real-Time Polymerase Chain Reaction / methods
  • Real-Time Polymerase Chain Reaction / standards
  • Sensitivity and Specificity
  • Telomerase* / genetics
  • Urinary Bladder Neoplasms* / genetics

Substances

  • Telomerase
  • TERT protein, human