Table 2.

Limitations of using next-generation sequencing (NGS) with circulating free (cf) nucleic acids extracted from blood samples at diagnosis of non-small cell lung carcinoma.

The quantity of cf-nucleic acid extracted from plasma samples may not be sufficient for NGS due to the tumor stage

Brain metastases usually shed a too low amount of tumor cf-nucleic acid into the blood for NGS

Some specific mutations in certain genes are associated with a low amount of cf-nucleic acid for NGS

Gene amplifications and rearrangements are less frequently detectable with cf-nucleic acid in blood samples as the same nucleic acid extracted from a tissue biopsy

Assessment of PD-L1 for first-line immune check point inhibitor treatment is not possible with blood samples

Pitfalls can be associated to NGS with cf-nucleic acid due to clonal hematopoiesis on circulating free germinal DNA

Validation and accreditation processes are more difficult to set up for NGS with blood samples than for NGS with a tissue biopsy