Towards a “Sample-In, Answer-Out” Point-of-Care Platform for Nucleic Acid Extraction and Amplification: Using an HPV E6/E7 mRNA Model System

Gulliksen, Anja; Keegan, Helen; Martín, C; O’Leary, John; Solli, Lars; Falang, I. M.; Grønn, Petter; Karlgård, Aina; Mielnik, Michał; Johansen, Ib-Rune; Tofteberg, Terje; Baier, Tobias; Gransee, Rainer; Drese, Klaus; Hansen‐Hagge, Thomas E.; Riegger, L.; Koltay, Peter; Zengerle, Roland; Karlsen, Frank; Ausen, Dag; Furuberg, Liv

doi:10.1155/2012/905024

Cited by 28 publications

(19 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The results matched well with the results of conventional NASBA techniques, but with only 1/2000 of the sample size needed compared to the conventional technique [42]. This technology was recently applied to detection of E6 and E7 mRNA, which is closely associated with HPV, on a POC platform [43]. The device is capable of performing pre-concentration, extraction, amplification, and fluorescent detection, and requires minimal user interface.…”

Section: A Optical Detection Technologiessupporting

confidence: 66%

Advances in Nanotechnology and Microfluidics for Human Papillomavirus Diagnostics

et al. 2015

View full text Add to dashboard Cite

| Human papillomavirus (HPV) has been shown in many studies as a prerequisite for the development of cervical cancer, which is the second most common and predominant form of malignancy among women worldwide, with 270 000 deaths (80% of whom live in developing countries) and 500 000 new cases per year. Early diagnosis of cervical cancer allows patients to be fully treated. Therefore, there is high clinical utility of HPV diagnostics even with binary positive/ negative indication of the presence of HPV in patient samples.Although the Pap smear is widely used for cervical cancer screening, this method still suffers from low sensitivity and specificity. Thus, simple, rapid and inexpensive diagnostic methods are needed to detect the etiology of cervical cancer, i.e., HPV, especially high-risk oncogenic subtypes 16 and 18.Here, we review the existing assays and platforms employed for HPV diagnosis, and highlight recent advances in nanotechnology and microfluidics that potentially enable new approaches for HPV diagnosis.

show abstract

Section: A Optical Detection Technologiessupporting

confidence: 66%

Advances in Nanotechnology and Microfluidics for Human Papillomavirus Diagnostics

et al. 2015

View full text Add to dashboard Cite

show abstract

“…cleavage by caspases) may also facilitate differentiation‐dependent genome amplification, with the accumulation of E1 in the nucleus enhancing viral DNA replication at the expense of cellular replication through induction of a DNA damage response . The E4 protein, which accumulates at very high levels in cells supporting virus synthesis may in fact have a primary function in virus release or transmission , with the optimization of genome amplification occurring as an indirect consequence of its expression .…”

Section: Papillomavirus Life Cycle Organization In the Infected Epithmentioning

confidence: 99%

Human papillomavirus molecular biology and disease association

Doorbar

Egawa

Griffin

et al. 2015

Reviews in Medical Virology

712

624

View full text Add to dashboard Cite

SummaryHuman papillomaviruses (HPVs) have evolved over millions of years to propagate themselves in a range of different animal species including humans. Viruses that have co‐evolved slowly in this way typically cause chronic inapparent infections, with virion production in the absence of apparent disease. This is the case for many Beta and Gamma HPV types. The Alpha papillomavirus types have however evolved immunoevasion strategies that allow them to cause persistent visible papillomas. These viruses activate the cell cycle as the infected epithelial cell differentiates in order to create a replication competent environment that allows viral genome amplification and packaging into infectious particles. This is mediated by the viral E6, E7, and E5 proteins. High‐risk E6 and E7 proteins differ from their low‐risk counterparts however in being able to drive cell cycle entry in the upper epithelial layers and also to stimulate cell proliferation in the basal and parabasal layers. Deregulated expression of these cell cycle regulators underlies neoplasia and the eventual progression to cancer in individuals who cannot resolve high‐risk HPV infection. Most work to date has focused on the study of high‐risk HPV types such as HPV 16 and 18, which has led to an understanding of the molecular pathways subverted by these viruses. Such approaches will lead to the development of better strategies for disease treatment, including targeted antivirals and immunotherapeutics. Priorities are now focused toward understanding HPV neoplasias at sites other than the cervix (e.g. tonsils, other transformation zones) and toward understanding the mechanisms by which low‐risk HPV types can sometimes give rise to papillomatosis and under certain situations even cancers. Copyright © 2015 John Wiley & Sons, Ltd.

show abstract

“…A pair of primers and a molecular beacon were designed to specifically amplify and detect the tmRNA transcript of H. influenzae. The primers (5'-3') P1; AATTCTAATACGACTCACTATAGGG-AGAAGG-CTTCGATCCTCAAACGGT, P2; GCAGCTTAACCT and a molecular beacon 5'FAM CCGAGT-GGGGATAACGCGGAGTCA-ACTCGG DAB 3' were designed according to recommended guidelines [33,34]. Primer and molecular beacon probes were supplied by Eurofins MWG Operon (Ebersberg, Germany).…”

Section: Nasba Assay Designmentioning

confidence: 99%

Development of an on-disc isothermal in vitro amplification and detection of bacterial RNA

Brennan

Coughlan

Clancy

et al. 2017

Sensors and Actuators B: Chemical

View full text Add to dashboard Cite

We present a centrifugal microfluidic "Lab-on-a-Disc" (LoaD) system capable of implementing nucleic acid in vitro amplification using non-contact heating and fluorescence detection. The system functionality is verified by implementing a Nucleic Acid Sequence Based Amplification (NASBA) reaction, targeting the tmRNA transcript of Haemophilus influenzae. The NASBA assay incorporates fluorescent molecular beacon probes reporting target tmRNA amplification for endpoint detection. The system implements non-contact IR heating to heat the NASBA reaction to the required target temperatures during denaturation and amplification steps. The LoaD control system facilitates spin speed and chamber positioning for heating and fluorescence detection. The LoaD alignment system uses magnetic fields to locate and lock the chamber in the required position (heating or detection). The NASBA assay was implemented on the system using Haemophilus influenzae tmRNA over the range 10 2-10 4 cell equivalent (CE) units. For comparison, identical qNASBA assays were implemented on a Roche LightCycler 2.0 over this concentration range.

show abstract

Towards a “Sample-In, Answer-Out” Point-of-Care Platform for Nucleic Acid Extraction and Amplification: Using an HPV E6/E7 mRNA Model System

Cited by 28 publications

References 30 publications

Advances in Nanotechnology and Microfluidics for Human Papillomavirus Diagnostics

Advances in Nanotechnology and Microfluidics for Human Papillomavirus Diagnostics

Human papillomavirus molecular biology and disease association

Development of an on-disc isothermal in vitro amplification and detection of bacterial RNA

Contact Info

Product

Resources

About