Highly Sensitive and Reliable Detection of EGFR Exon 19 Deletions by Droplet Digital Polymerase Chain Reaction.
Mol Diagn Ther
; 21(5): 555-562, 2017 10.
Article
en En
| MEDLINE
| ID: mdl-28589341
ABSTRACT
BACKGROUND:
Analysis of EGFR mutations is becoming a routine clinical practice but the optimal EGFR mutation testing method is still to be determined.METHODS:
We determined the nucleotide sequence of deletions located in exon 19 of the EGFR gene in lung tumor samples of patients residing in different regions of Russia (153 tumor DNA specimens), using Sanger sequencing. We developed a droplet digital polymerase chain reaction assay capable of detecting all common EGFR deletions in exon 19. We also compared the therascreen amplification refractory mutation system assay with a droplet digital polymerase chain reaction assay for the detection of all the deletions in our study.RESULTS:
The droplet digital polymerase chain reaction assay demonstrated 100% sensitivity against polymerase chain reaction fragment length analysis and detected all possible types of deletions revealed in our study (22 types). At the same time, the therascreen EGFR RGQ PCR Kit was not able to detect deletions c.2252-2276>A and c.2253-2276 and showed low performance for another long deletion.CONCLUSION:
Thus, we can conclude that the extraordinary length of deletions and their atypical locations (shift at the 3'-region compared to known deletions) could be problematic for the therascreen EGFR RGQ PCR Kit and should be taken into account during targeted mutation test development. However, droplet digital polymerase chain reaction is a promising and reliable assay that can be used as a diagnostic tool to genotype formalin-fixed paraffin-embedded cancer samples for EGFR or another clinically relevant somatic mutation.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Reacción en Cadena de la Polimerasa
/
Eliminación de Secuencia
/
Carcinoma de Pulmón de Células no Pequeñas
/
Receptores ErbB
/
Neoplasias Pulmonares
Tipo de estudio:
Diagnostic_studies
/
Observational_studies
Límite:
Female
/
Humans
/
Male
País como asunto:
Asia
/
Europa
Idioma:
En
Año:
2017
Tipo del documento:
Article