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HPV E6/E7 mRNA Expression Analysis Using the NovoCyte® Flow Cytometer

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Enzo Life Sciences Farmingdale, NY USA
ACEA Biosciences, San Diego, CA USA

Featured Product: FLOWSCRIPT® HPV E6/E7 assay kit


INTRODUCTION

Human Papillomavirus (HPV) is the most common sexually transmitted disease and the chief cause of cervical cancer. HPV infects cervical tissues as well as other mucosal and cutaneous regions of the body. Primarily, HPV infection is asymptomatic and transient but it may persist and cause cervical cellular abnormalities. The primary tool for cervical cancer screening, the Pap smear, determines the presence of dysplasia of the cervical squamous epithelium. Widespread screening has been extremely effective for the treatment and prevention of cervical cancer, however, cytology methods of cervical screening require highly trained cytopathologists and do not diagnose HPV infection. Since the discovery of HPV as the underlying cause of cervical cancer, morphologically abnormal cervical cells have been routinely screened for the presence of HPV DNA by PCR analysis. HPV DNA analysis in cervical cancer screening has increased the detection of high-risk lesions with normal cytology results and decreased unnecessary biopsies in women with atypical squamous cells of undetermined significance (ASC-US).

Fluorescent in situ hybridization (FISH) is a powerful technique used for the detection of RNA or DNA in intact cells with a fluorescent probe. Recently FISH technology has been combined with flow cytometry to allow for rapid examination of RNA expression in individual cells. Detection of viral HPV mRNA instead of DNA is a more direct indication of viral activity; therefore the detection of HPV RNA is clinically useful for HPV detection. Overexpression of two HPV oncoproteins E6 and E7 contribute to the malignancy of HPV infected cells by inhibition and degradation of tumor suppressor proteins and promoting cell growth. With new innovations in FISH and flow cytometry, it is possible to detect the presence of E6/E7 HPV mRNA with the use of the FLOWSCRIPT HPV E6/E7 assay by Enzo Life Sciences and the NovoCyte flow cytometer providing clinically relevant information at the single cell level. To demonstrate the ability of HPV mRNA detection on the NovoCyte Flow cytometer, both HPV control cell lines and clinical samples were assayed for E6/E7 mRNA transcription and analyzed on the NovoCyte flow cytometer.

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