Impact of Fine Needle Aspiration Collecting Medium on Success of Polymerase Chain Reaction Sequencing
Key Clinical Takeaways
- Design/Population: As presented at the 2025 American Society for Clinical Pathology (ASCP) Annual Meeting in Orlando, Florida, a prospective cohort study of 24 FNA specimens from advanced NSCLC compared CytoRich Red (n = 12) and formalin (n = 12) as collection media for PCR sequencing of KRAS and EGFR mutations.
- Key Outcomes: CytoRich Red produced higher DNA yields (1.1 to 43.5 ng/μl) and 11 of 12 concordant results, vs 0.6 to 10.4 ng/μl and 6 of 12 concordant results with formalin.
- Clinical Relevance: CytoRich Red demonstrated superior performance as a collecting medium for PCR sequencing; squamous cell carcinoma specimens yielded lower DNA quality than non-squamous cases.
According to a small cohort study, CytoRich Red performed better than formalin as a collecting medium for polymerase chain reaction (PCR) sequencing among patients with advanced non-small cell carcinoma (NSCLC).
These data were first presented by Adeyinka Akinsanya, MD, Indiana University, Indiana, at the American Society for Clinical Pathology 2025 Annual Meeting in Orlando, Florida.
Dr Akinsanya and coauthors explained that pre-analytic factors, such as the collecting medium form fine needle aspiration (FNA) rinse and the portion of sample used in sequencing, “may influence the success rate of molecular sequencing.”
This prospective study included 24 FNA specimens from advanced NSCLC. Specimens were collected in either CytoRich Red (n = 12) or formalin (n = 12). DNA yield and PCR sequencing outcomes for KRAS or EGFR mutations were evaluated. The normal process of obtaining samples during rapid on-site evaluated was modified for this study, to include an additional designated pass obtained and placed in appropriate media to test the supernatant.
Of the 12 samples in the CytoRich Red group, 10 were adenocarcinoma and 2 were NSCLC cases. All samples yielded satisfactory DNA quantities (1.1 ng/μl to 43.5 ng/μl), and 11 samples yielded results concordant with positive controls. Of the 12 samples in the formalin group, 7 were squamous cell carcinomas, 2 were adenocarcinomas, 1 was NSCLC, and 1 was large cell neuroendocrine carcinoma. DNA concentrations in this group ranged from 0.6 ng/μl to 10.4 ng/μl, and 9 specimens generated with valid results with 6 concordant with positive controls.
Dr Akinsanya et al concluded that CytoRich Red “demonstrated superior performance as a collecting medium for PCR sequencing.” They also noted squamous cell carcinoma specimens yielded lower DNA quality when compared to non-squamous cell carcinoma lung cancer cases.
Source:
Akinsanya A, Madison R, Gorden R, et al. Use of fine needle aspiration supernatant from lung carcinoma for PCR sequencing: Does the collecting medium make a difference? Presented at ASCP 2025 Annual Meeting; November 17-20, 2025. Orlando, Florida.
