The Epi proLung BL Reflex Assay was validated in a case control study conducted for registration of the IVD as a CE-marked test. In this performance evaluation study the following performance characteristics were determined:

Clinical Sensitivity and Specificity. The performance of the biomarker ^mSHOX2 was established in a performance evaluation study for CE-marking of the Epi proLung BL Reflex Assay with samples from 75 patients with lung cancer and 79 patients with other lung diseases (e. g. infections, sarcoidosis, scleroderma, COPD). The ^mSHOX2 assay consistently identified cancer cases with a sensitivity of 81 % at a specificity of 95 %.

Robustness. The Epi proLung BL Reflex Assay uses routinely obtained, fixed sample material with no additional preparation required. Handling and ease-of-use of the Epi proLung BL Reflex Assay has been further optimized by combining as many test components as possible in the fewest number of reaction vessels possible. Moreover, major handling failures that could lead to false negative or false positive results were analyzed. It was verified that these failure modes were mitigated by the test design or detected by the work flow controls.

Reproducibility. Several aliquots of identical samples were analyzed repeatedly with the Epi proLung BL Reflex Assay. This study was performed according to guidance from the CLSI (Clinical and Laboratory Standards Institute) to demonstrate reproducibility of test results. Three different pools of bronchial lavage samples as well as mixtures of DNA with known relative amounts of methylation were analyzed. Three different reagent lots were used and the samples were analysed by three different operators on three different real-time PCR systems. The Epi proLung BL Reflex Assay was shown to deliver consistent results.