PCR-ISH application potentiates the clinical and diagnostic assessment of cervical tissue.

R. Li Vigni PhD1,2, U.A Bianchi MD2, G. Carosi MD3, M. Lomini MD2, S. Pecorelli MD2.

1Institut Pasteur, Paris, France; 2Department of Obstetrics and Gynaecology and 3Department of Infectious and Tropical Diseases, University of Brescia, Spedali Civili of Brescia, Italy.

AIM: The emergence of molecular techniques has enormously contributed to etiologically link human papillomavirus (HPV) infections to cervical disease but, so far, no general agreement exists on a choice method for HPV detection. Nowadays, the diagnosis of cervical pathology remains sometimes problematic. This study analyse the diagnostic value of in-situ hybridization (ISH) after gene amplification by polymerase chain reaction (PCR-ISH). METHODS: All patients were undergone to colposcopic-directed biopsy after a diagnosis of cytological abnormalities. We compared 100 women infected with human immunodeficiency virus type-1 (HIV+) and 100 HIV- women with similar age for: i) the prevalence of cervical intraepithelial neoplasia (CIN); ii) HPVs detection by ISH, PCR-ISH and solution-phase PCR from the different categories of histological specimens. HPV detection was performed in parallel analyses by ISH and PCR-ISH as previously described (Li Vigni et al, 1999) and by standard PCR using MY09/11 primers; amplimers therefrom produced were revealed by blot-hybridization. RESULTS: The prevalence of CINs ranged from 46 for HIV- to 68% for HIV+ women. The cumulative HPV positivity was higher for HIV+ than HIV-women. In both cohorts, 59% of lesions were found ISH-positive. Viral sequences were discovered in about 100% of CIN2-3 both by PCR and PCR-ISH. On the whole, 89% and 80% of condylomas were positive by either PCR or PCR-ISH in respectively HIV- and HIV+ patients. CIN1 harboured HPVs in 75% and 81% of lesions by PCR in HIV- and HIV+ women whereas 83% and 87% were found positive by PCR-ISH. Additionally, PCR-ISH evidenced HPV sequences in 25% and 33% of chronic cervicitis in HIV- and HIV+ patients. CONCLUSIONS: The diagnosis and classification of cervical disease is sometimes a challenge for the pathologist. In fact, the use of cyto-histopathological criteria may result difficult, subjective and poorly reproducible especially when borderline or low-grade lesions are evaluated. Because the PCR-ISH offers the opportunity to specifically probe tissues with little morphological changes such information is of particular value for selecting true high-risk patients inside groups at-risk for cervical carcinoma.

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Paper presented at the International Symposium on Predictive Oncology and Intervention Strategies; Paris, France; February 9 - 12, 2002; in the section on Risk Assessment, Part 1.