The studies presented above were aimed to index the CVF proteome, and reveal the remarkable proteins for malign transformations of the cervix. Yet, there are 2 studies which performed quantitative validation of potential biomarkers aimed at the development of a new screening method for CxCa. FirstThe first of these two studies; by Van Raemdonck et al. (2014) [59] aimed theto illustrate the different protein expression profile of the CVF fromin healthy and in infected women. In this direction, they performed a semi-quantitative proteomics study. Cervicovaginal lavage samples were collected during the colposcopic examination by aspiration, with a 5% acetic acid, from 6 healthy, 4 LSILLSIL-, 2 HSIL diagnosedHSIL-diagnosed women. ProteomicsThe proteomics analysis was performed with a High Performance Liquid Chromatography (HPLC) and a Matrix Assisted Lazer Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF/TOF). They found that 16 proteins significantly differ statistically between the stages statistically significant. In particular, they stated that actinin-alpha-4 (ACTN4; p: 0,001) as the most promising protein is the potential biomarker and suitable for developing ofthe POC test for the CxCa screening. To demonstrate this opinion as quantitatively, the team also performed an Enzyme-Linked ImmunoSorbent Assay (ELISA) test from 16 healthy and 12 HPV-infected precancerous samples, and the results confirmed the performance of ACTN4. The Area Under Curve (AUC) of the Receiver Operating Characteristic (ROC) was 86%, with 84% of sensitivity and 86% of specificity.
Proteomics studies with the latest-technology devices can expand the CVF proteome and reveal novel candidate biomarkers. WhenNow that we have reviewed these studies, currently we have several potential candidate protein biomarkers that neededneed to be validated and verified by further clinical studies. The development of an immunoassay algorithm or aan integrated multiplex microfluidic cancer biochip with two or three biomarkers with the highest diagnostic discrimination will be one of the best state-of-the-art achievements of our time.