BACKGROUND: Despite the fact that in situ hybridization (ISH) has been extensively studied, there have been limited reports of the usefulness of this technique in malignant transformation and HPV-DNA integration in relation to histological findings (24. De Marchi Triglia R. 2009, 26. Ming Guo. 2008, 31. Zappacosta R 2013)
OBJECTIVE: To assess the usefulness of HPV genotyping using in situ hybridization ISH technique to identify episomal and punctate signal pattern and to verify whether a punctate pattern could be used as a progression marker of CIN
METHODS: The study period was between June 2013 through July 2014. HPV status was determined in 40 ever-married women (25 - 53 years age) using in situ hybridization, 20 women (group I) with healthy looking cervices and normal Pap smear, and 20 women (group II) with abnormal cervices and abnormal Pap smears. A third group, (group III), 20 women with squamous cell carcinoma. Pap smears were collected, followed by colposcopic guided biopsy. HPV was tested on all specimens by in situ hybridization (ISH) using the broad spectrum HPV probe recognizing HPV 16, 18, 31, 33, ,35,39, 45, 51, 52, 56, 58, 59, and 68 compared with conventional hematoxylin ad eosin H&E stain
RESULTS: 4 of the 20 women (group I) showed mild positive ISH test results with diffuse nuclear staining. Group II results; mild positive reaction with diffuse nuclear signal pattern was noticed in 4 (20%) of the cervical specimens. Moderate positive reaction with diffuse signal pattern of the nucleus was observed in 8 (40%) of the specimens. The remaining 8 (40%) of the specimens showed both diffuse and punctate signals in the nuclei.
CONCLUSION: ISH is a sensitive, easy to handle method for HPV-detection in cervical squamous intraepithelial lesions and cancers, suitable for routine pathology, helps to distinguish episomal from integrated HPV, and hence, prognosis of the lesion.
References
24. De Marchi Triglia R., Metze K, Zeferino LC, Lucci De Angelo Andrade LA: HPV in situ hybridization signal patterns as a marker for cervical intraepithelial neoplasia progression. Gynecol Oncol. 2009;112:114–118. (Abstract)
26. Ming Guo, Yun Gong, Michael Deavers, Elvio G. Silva, Yee Jee Jan, David E. Cogdell, Rajyalashmi Luthra, E. Lin, Hung Cheng Lai, Wei Zhang, and Nour Sneige, Evaluation of a commercialized In situ hybridization assay for detecting human papillomavirus DNA in tissue specimens from patients with cervical intraepithelial neoplasia and Cervical Carcinoma, Journal of Clinical Microbiology, Jan. 2008, p. 274–280 Vol. 46, No. 10095-1137
31. Zappacosta R, Antonella Colasante, Patrizia Viola, Tommaso D’Antuono, Giuseppe Lattanzio, Serena Capanna, DanielaMaria Pia Gatta, and Sandra Rosini: Chromogenic in situ hybridization and p16/Ki67 dual staining on formalin-fixed paraffin-embedded cervical specimens: correlation with HPV-DNA test, E6/E7 mRNA test, and potential clinical applications. BioMed Research International. Volume 2013, Article ID 453606, 11 pages
Key words: Cervical intraepithelial neoplasia CIN, in situ hybridization ISH, human papillomavirus HPV