Adrian Rea¹, Courtney Noah¹, Andrew L’Huillier¹, Jack Coleman¹, Michael Mira², and Dieter Schapfel^2
1Enzo Life Sciences, Farmingdale, NY 11735
²Enzo Clinical Labs, Farmingdale, NY 11735

Featured Product: FLOWSCRIPT® HPV E6/E7 assay kit

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Abstract

Enzo’s newest flow cytometry platform, FLOWSCRIPT®, acts as “a snapshot cell query” by allowing the analysis of mRNA transcript expression in individual cells in a mixed cell population. By studying whether a gene or a set of genes is turned on or off, it is possible to obtain clinically relevant information at the single cell level. The first product developed for use with this technology is the FLOWSCRIPT® HPV E6/E7 Assay. Integration and ultimately overexpression of HPV oncoproteins, E6 and E7, promotes the growth of malignant cells through the inhibition of tumor suppressors and has been linked with increased likelihood of cervical cancer progression.

Introduction

The FLOWSCRIPT® HPV E6/E7 Assay is a flow cytometry-based assay for the detection of mRNAs that precede the expression of the oncogenic proteins, E6 and E7, produced during infection by high-risk Human Papillomavirus (HPV) viruses. The FLOWSCRIPT® HPV E6/E7 Assay is capable of detecting E6/E7 mRNA transcripts from multiple high-risk HPV types (HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 82) which together account for over 95% of cervical cancer. The assay employs a novel in situ hybridization technique utilizing cocktails of oligonucleotide probes specific to multiple targets within the E6 and E7 genes to ensure the detection of these transcripts from most known variants of high-risk HPV. Fixed and permeabilized cells are hybridized with probes and analyzed by flow cytometry for E6/E7 transcript expression. Each probe contains a fluorescent label and a quenching molecule whereby no signal is observed in the absence of target. During hybridization, the probe anneals to the target sequence, thereby emitting a detectable signal.