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PCR-ISH application potentiates the clinical and diagnostic assessment of cervical tissue.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. For more information, contact livigni@pasteur.fr 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. |
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