Achievements
Claudin 1 as a biomarker for cervical cytology and histology
(EUROGIN, Montecarlo, 17-20 February, 2010)
In cooperation with:
GenoID Ltd., Budapest, Hungary; Semmelweis University, Dept. of Pathophysiology, Budapest, Hungary; Semmelweis University, 2nd Department of Obstetrics & Gynecology Budapest, Hungary; Semmelweis University, Dept. of Microbiology, Budapest, Hungary
The claudin1 (CLDN1) is major component of the tight junction structure and plays an important role in cell-cell adhesion. The role of CLDN1 in cervical cancer pathology was revealed by our previous work (2). In this study 360 cervical cytological, histological samples were collected from a colposcopic referral population. The histological and cytological samples were immunostained using CLDN1, and as reference, p16 INK4a /CDK2A antibodies and were evaluated by experienced pathologist.
The results indicate high concordance between CLDN1 and CDKN2A immunostaining, which were in agreement with histology immune status, as well. CLDN1 might have a similar potential, than CDKN2A, to be used as histological/cytological biomarker to improve the clinical performance of cervical cytology/histology.
Work was supported by FP7 Autocast 201525 and NKTH Jedlik Ányos SPE_SAFE.
Evaluation of a new cervical screening biomarker panel
(International Papillomavirus Congress, Malmö, 8-14 May, 2009)
In cooperation with:
GenoID Ltd., Budapest, Hungary; Semmelweis University, Budapest, Hungary; St. István Hospital, Budapest Hungary; Bajcsy-Zsilinszky Hospital, Budapest, Hungary; St. Lázár Hospital, Salgótarján, Hungary; Computer and Automation Research Institute, Hungarian Academy of Sciences, Hungary
Primary HPV screening is an emerging concept, which may fundamentally influence cervical cancer screening, making impossible to establish new high throughput automated screening methods in primary screening. Following this path, the combination of HPV testing with screening biomarkers could be the next major milestone. With the advent of a fully automated, low cost, biochemical screening method both the overall quality of the cervical screening programs could be improved and also the programs could be more accessible for women, especially in low resource countries.
We have developed a new gene expression, HPV combination panel for cervical screening. The genes were identified using TaqMan® Custom Array on 293 patient sample, detecting expression of 190 preselected genes in duplicates (the overall number of screening PCR reactions were 114,000). Thorough analysis revealed a classificatory system which identified 8 marker genes, which were capable to produce ROC area better than 0.9 and classify the samples with high precision. The method was validated on a colposcopic referral population using liquid-based cytological specimens, giving 95% specificity and 80% sensitivity for CIN1+ histology.
Work was supported by EC FP7 Autocast project and Hungarian Jedlik Ányos HPV_SCREEN and SPE_SAFE state grants.
High throughput HPV screening using a one step multiplex real-time PCR based system
(International Papillomavirus Congress, Malmö, 8-14 May, 2009)
In cooperation with:
GenoID Ltd., Budapest, Hungary; Semmelweis University, Dept. of Pathophysiology, Budapest, Hungary; Dept. of Histopathology, University of Dublin, Trinity College and The Coombe Women and Infants University Hospital, Dublin 8, Ireland; Semmelweis University, Dept. of Microbiology, Budapest, Hungary
Primary HPV screening is an emerging concept, which may fundamentally influence cervical cancer screening ( J. Cuzick et al. Vaccine 26S (2008) K29–K41). To fulfill the demand we have developed a molecular beacon based one step multiplex real-time PCR (MB-RT PCR) system that is designed for the high throughput detection of 14 high-risk HPVs (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68) on the ABI7900HT instrument, in 96 or 384 well plate format. Detection is achieved in 3 dye channels. Recognizing the demand for the identification of HPV vaccine types, HPV 16 and 18 are detected together in one channel. The other high-risk types are detected in groups separately and an artificial internal control is also detected to control for false negative reactions.
Using two patient populations; a cervical screening population and a colposcopy referral population, the performance of the MB-RT PCR was compared to cytology, histology, Full Spectrum HPV Amplification and Detection System (GenoID, IVD-CE mark), Amplicor HPV Test (Roche) and Hybrid Capture II (hc2, Qiagen). We propose the MB-RT PCR is a useful tool for high throughput HPV based cervical screening. In addition this technology has the capability for semiquantitative viral load determination which could potentially be used to improve specificity of primary HPV screening.
Work was supported by EC FP7 Autocast project and Hungarian Jedlik Ányos HPV_SCREEN and SPE_SAFE state grants.
