mRNA and DNA Detection of Human Papillomaviruses in Women of All Ages Attending Two Colposcopy Clinics

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From: PLoS ONE(Vol. 7, Issue 11)
Publisher: Public Library of Science
Document Type: Article
Length: 4,626 words
Lexile Measure: 1560L

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Author(s): Aris Spathis 1 , * , Christine Kottaridi 1 , Aikaterini Chranioti 1 , Christos Meristoudis 1 , Charalambos Chrelias 2 , Ioannis G. Panayiotides 3 , Evangelos Paraskevaidis 4 , Petros Karakitsos 1

Introduction

Human Papillomavirus (HPV) infection is estimated to be the most common sexually transmitted infection [1] and has been directly linked to cervical intraepithelial lesion creation and increased risk of cervical cancer development [2], [3]. Although HPVs are commonly detected in cervical smears, most infections appear to be transient and asymptomatic, with approximately 90% of infections being cleared within 2 years [4], [5], [6]. Only persistent HPV infections seem to be linked to CIN establishment and progression [5], [7], [8].

Although the life-long risk for HPV infection is 80%, only a fraction of that percentage of women will eventually develop cervical cancer [4], [9]. Furthermore, HPV DNA positivity usually peaks in younger women, ranging from 25%-65% with a steady decline in women older than 35 years, ranging from 10%-30% [4], [6], [10], [11]. As a result HPV DNA testing has been suggested for screening of women over 30 or women with equivocal cytology results [12], [13].

The expression of the two viral genes E6 and E7 is responsible for the transformation of cells by HPV, resulting in continuous morphological changes and finally cervical neoplasia [14], [15], [16], [17], [18]. Therefore, detection of HPV oncogene transcripts has been proposed to identify more accurately women with higher grade lesions and increased risk of cervical cancer development [19], [20], [21], [22], [23].

In the present study the clinical performance of flow cytometric in-situ hybridization for E6 and E7 mRNA transcript detection was evaluated, in women attending two colposcopy clinics of two University Hospitals in Greece.

Materials and Methods

Study Design

A total of 1173 women that attended the colposcopy clinics of the 3 rd Department of Gynecology and Obstetrics in the tertiary care "ATTIKON" University General Hospital and the Department of Gynecology and Obstetrics in the University Hospital of Ioannina were enrolled in the present study. The study population did not represent a normally screened population, since most patients attended the outpatient clinics after a referral abnormal cytology and or colposcopy. All patients signed an informed consent form, while the study was approved by the Bioethics comity of the hospitals. A liquid-based cytology (LBC, ThinPrep® Pap-Test, Hologic, U.S.A.) sample was collected and a monolayer smear was prepared on a TP 2000 Processor and stained according to Papanicolaou technique. A trained cytopathologist diagnosed each case, according to the Bethesda 2001 system [24]. Biopsies were taken from all women with either colposcopically visible lesions or a cytology result of ASCUS or worse. Tissue samples were routinely processed and diagnosed by an experienced pathologist. Two 1 ml LBC aliquots were used for DNA extraction and flow cytometric analysis.

HPV DNA Detection

HPV DNA detection was performed on the first aliquot using a commercially available kit, CLART® HPV 2 kit (Genomica, Spain) that allowed the detection of 35 different HPV genotypes, 20...

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Gale Document Number: GALE|A477082536