GYNECOLOGIC ONCOLOGY 46, 33-36 (1992) The Prevalence of HPVI 6 DNA in Normal and Pathological Cervical Scrapes Using the Polymerase Chain Reaction NINO PASETTO,* FRANCESCO SESTI,* LAVINIA DE SANTE,* EMILIO PICCIONE,~ GIUSEPPE NOVELLI,*Y~?’ AND BRUNO DALLAPICCOLA$ *Clinica Ostetrica e Ginecologica, Universitri di Roma Torvergata 00173 Rome, Italy; TCattedra di Patologia Ostetrica e Ginecologica, Universitri di Cagliari, 09100 Cagliari, Sardinia, Italy; and Kattedra di Genetica Umana, Universitk di Roma Torvergata e Universitk di Urbino, Urbino, 61029 Italy Received May 16, 1991 Human papillomavirus (HPV) has beenimplicated in the de- velopment of cervical cancer. Newly developed moleculartech- niques have facilitated I-IPV identification and typing of cervical lesions. Several studieshave suggested a strong correlation be- tween HPV16 and intraepithelial cervical neoplasia.However, there is no agreement on this point sinceHPV16 is detectedalso in normal cervical tissue. We have amplified the Ll region of HPV16 DNA using the polymerase chain reaction (PCR) tech- nique and analyzedthe HPV16 prevalence in cytological scrapes of 201 Italian sexualIyactive patients. One hundred twenty-four of these women had normal cytologic and colposcopic exami- nation, while seventy-seven womenhad normal or abnormal cy- tology and a cervical lesion detectable by colposcopy. Within this latter group, 25 of 77 women (32.4%) were positive for HPV16 DNA, compared to 11 of 124samples (8.87%)from women with normal cytology and colposcopy. The present results confirm the high efficiency of the PCR techniquein detecting HPV16 DNA from cervical scrapes and suggest that this analysis should become a method of choice in clinical studies. o 1s~ ~cad~nic prrss, IK. INTRODUCTION Newly developed molecular techniques have allowed the identification and detection of 60 different human papillomavirus (HPV) types. More than 20 of them have been found in the genital tract [ 11. HPVs have been class- ified as low risk and high risk types according to their detection in either benign condylomas/papillomas or cervical intraepithelial neoplasia (CIN) [2,3]. Evidence has indicated a strong correlation between HPV16 and the development of cervical cancer [4]. Nevertheless, re- cent studies have reported a high frequency of HPV16 DNA in normal cervical tissue [5]. This study was devised ’ TO whom correspondence should be addressed at L&oratorio Cen- trale, C.R.I. - Servizio di Genetica, Via Ramazzini 15, 00151 Rome, Italy. Fax: 39-6-65741183. to assessthe prevalence of HPV16 in cervical scrapes from a group of Italian women undergoing cytologic, histologic, and colposcopic examinations. This has allowed evalua- tion of the rate of HPV16 infection in normal and ma- lignant cervical tissue. MATERIALS AND METHODS Two hundred one sexually active patients (age range, 19-65 years) sent to us for further evaluation after suspect clinical observation of cervical epithelium during the 3 months from November 1990 until 31 January 1991 were subjected to colposcopy. Seventy-seven patients had a cervical lesion, seventy-two had an atypical transforma- tion zone (ZTA) (mosaic, punctation, acetowhite epithe- lium), and five had a colposcopically suspected invasive carcinoma. Nine of the seventy-seven patients with cerv- ical lesions had concomitant HPV alterations of the vagina and/or vulva. In 1 case an intraepithelial neoplasia of the vagina (VAIN 3) was present. The other 124 women had normal colposcopic examination, in the absence of vaginal or vulvar lesions. The colposcopic examination was per- formed according to Reid’s colposcopic index [6]. All the patients with ZTA and the 5 women with suspected in- vasive carcinoma were submitted to a cervical target punch biopsy. The severity of the CIN was graded as CIN 1, 2, and 3, according to the criteria of Ferenczy and Winkler [7]. A cytohistological diagnosis of HPV infection was recorded only in the presence of koilocytes. Two cervical smears were obtained from each of the 201 pa- tients. Thirty-four patients with unsuccessful colposcopy were submitted to an endocervical scrape. One smear was fixed on a slide for routine Papanicolaou examination and assessed for features of CIN [8] and for evidence of HPV infection [9]. The second cytologic smear was used for 33 0090~8258/92 $4.00 Copyright 0 1992 by Academic Press,Inc. All rights of reproduction in any form reserved.