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  4. Human Papillomavirus DNA and Host Gene mRNA Biomarker Detection in a "Micro Total Analysis System" (microTAS) Device to Aid Diagnosis of Cervical Intraepithelial Neoplasia
 
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2014
Journal Article
Title

Human Papillomavirus DNA and Host Gene mRNA Biomarker Detection in a "Micro Total Analysis System" (microTAS) Device to Aid Diagnosis of Cervical Intraepithelial Neoplasia

Title Supplement
Abstract
Abstract
Background: Human papillomavirus (HPV) DNA detection has high sensitivity for high-grade cervical intraepithelial neoplasia (CIN), but low specifi city. Cervical cancer research is focused on development of biomarkers (BM) for prediction of disease severity eg. HPV E6/E7mRNA and p16INK4a/Ki67. This study describes a novel set of host mRNA biomarkers, mediator probe PCR (MP-PCR), and microTAS device for HPV DNA and BM detection in LBC. Design: A panel of 21 BM were validated by TaqMan PCR assays on 692 women with known histology and HPV status (Hybrid Capture 2 (HC2) and Linear Array HPV Genotyping (LA-HPV)) recruited from colposcopy clinics through CERVIVA (The Irish Cervical Screening Research Consortium) and Jedlik Anyos HPV Screen Multicentric Clinical Study, Hungary. A novel MP-PCR test was designed for HPV and BM detection, clinically validated on AB7900 thermal cycler and miniaturised for use in a microfluidic chip run on an instrument designed for multichannel thermal cycling and fluorophore detection (microTAS device). Results: Combined BM and HPV testing by TaqMan and hc2 had high specificity (91%), and sensitivity (75%) for CIN1+. HPV16 MP-PCR on AB7900 had high positive predictive value (95%), comparable to HC2 (94%) for CIN2+. On the microTAS device, the HPV16 MP-PCR had a limit of detection of 160 HPV16 copies with high sensitivity (85%) and concordance (82%) to LA-HPV for HPV16 detection. Concordance of TaqMan PCR and the microTAS device for 4 of the mRNA BM ranged from 79% to 83.3%. Conclusions: Combined mRNA BM and HPV DNA detection by PCR methods may be a useful alternative to consecutive BM and HPV, can be readily adapted for microTAS and should be explored further.
Author(s)
Keegan, H.
Pilkington, L.
Jordan, R.
Mozes, J.
Varga, N.
Benczik, M.
Riegger, L.
Koltay, P.
Sauer, U.
Preininger, C.
Koger, B.
Fraunhofer-Institut für Physikalische Messtechnik IPM  
Brandenburg, A.  
Fraunhofer-Institut für Physikalische Messtechnik IPM  
Muller, W.
Faltin, B.
Roth, C.
Martin, C.
Jeney, C.
O'Leary, J.J.
Journal
Laboratory investigation  
Conference
United States & Canadian Academy of Pathology (USCAP Meeting) 2014  
Language
English
Fraunhofer-Institut für Physikalische Messtechnik IPM  
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