Multigene Combined Detection by RT-qPCR Using Cytological Specimens.

Area of Science:

• Oncology
• Molecular Biology
• Genetics

Background:

• Lung adenocarcinoma is a major cause of cancer-related deaths.
• Accurate molecular profiling of driver gene mutations is crucial for targeted therapy selection.
• Cytological specimens offer a minimally invasive source for molecular testing.

Purpose of the Study:

• To evaluate the efficiency of the disposable membrane cell collector method (MCM) versus the cell block method (CBM) for DNA/RNA extraction from cytological specimens.
• To investigate the mutation status of ten key driver genes in advanced lung adenocarcinoma.
• To determine the clinical significance of these mutations in relation to patient demographics.

Main Methods:

• 155 advanced lung adenocarcinoma cytological specimens were analyzed.
• DNA/RNA extraction was performed using either the MCM (47 cases) or CBM (108 cases).
• Ten driver genes (EGFR, ALK, ROS1, BRAF, KRAS, NRAS, HER2, RET, PIK3CA, MET) were detected using amplification refractory mutation system and RT-qPCR.

Main Results:

• MCM yielded significantly higher RNA (p = 0.005) and DNA (p = 0.001) purity compared to CBM.
• MCM achieved a 100% success rate for multigene detection, while CBM had a 5.6% failure rate.
• EGFR (57.71%), ALK (6.04%), and ROS1 (3.36%) were the most frequent mutations. Mutation status correlated with gender and age.

Conclusions:

• MCM is superior to CBM for DNA/RNA extraction and multigene detection in cytological specimens.
• EGFR, ALK, and ROS1 mutations are prevalent in advanced lung adenocarcinoma.
• Further research is needed to understand and target coexisting mutations.