Sequenziamento mediante pyrosequencing: un’analisi rapida e sensibile per la determinazione delle mutazioni di EGFR nel carcinoma polmonare non a piccole cellule
Lung cancer is the most common tumor and one of the most aggressive, with rates of survival at 5 years of about 10% in most countries. Studies reported EGFR (Epidermal Growth Factor Receptor) gene activating mutations in 10–15% of NSCLC (Non Small Cell Lung Cancer) in the European population.
Molecular assessment of EGFR mutations became mandatory for gefitinib administration in NSCLC patients. Only tumors harboring at least one activating mutation in either exon 18, 19 or 21 are eligible for therapy with this drug.
Several methods are available in order to study EGFR mutations, such as direct sequencing and Real Time PCR. Pyrosequencing is a DNA sequencing technique based on the principle of sequencing-by-synthesis and it is characterized by a greater sensitivity compared to direct sequencing. We compared pyrosequencing with Real Time PCR, both combined with CE-IVD approved commercially available kits for EGFR analysis, in terms of effectiveness, economic and logistic impact.
In conclusion, although Real Time PCR has a greater sensitivity, pyrosequencing is an effective and reliable method, with an advantage in detecting new mutations whose clinical impact is yet unknown. More studies are needed in order to define which sensitivity level is really correlated to clinical response.
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