Double staining: diagnostic utility in non-small cell lung carcinoma in the era of tissue conservation.

Background: In an era of precision medicine distinguishing pulmonary squamous cell carcinoma (SQCC) from adenocarcinoma (ADC) is vital for treatment. Immunohistochemical (IHC) staining for p40, p63 and Cytokeratin 5 (CK5) are useful for SQCC, while TTF-1 and Napsin-A can be used for confirming ADC. Fine needle aspiration (FNA) cell blocks (CB) have limited tissue, hence, double IHC staining is helpful for tissue conservation for molecular analysis.

Methods: Thirty six confirmed lung SQCC and 45 ADC CB were selected for IHC. Double staining was performed with p40/CK5 and p63/CK5 on all SQCC, and with TTF-1/Napsin-A on all ADC. Results were positive if at least 5% of malignant cells were immunoreactive for the antigen.

Results: P40/CK5 had (92%) sensitivity, (100%) specificity, (100%) positive predictive value (PPV), (91%) negative predictive value (NPV) and an overall diagnostic accuracy of (96%). By contrast, P63/CK5 double stains showed (92%) sensitivity, (80%) specificity, (85%) PPV, (89%) NPV and (86%) overall diagnostic accuracy, respectively. TTF-1/Napsin A staining for ADC showed a sensitivity of 80%, specificity of 96%, PPV of 97%, NPV of 71% and accuracy of 85%.

Conclusions: P40/CK5 double stain has higher specificity, PPV, NPV, and overall accuracy than P63/CK5 double stain in the diagnosis of lung SQCC. TTF-1/Napsin-A double staining is a valuable marker with high specificity, PPV, and diagnostic accuracy in diagnosing lung ADC. The usage of P40/CK5 and TTF-1/Napsin-A as a panel can be recommended for characterizing non-small cell carcinoma (NSCC) of the lung and for conserving tissue for molecular testing.