Standardization of the chromogenic in situ hybridization protocol for searching the high and low risk HPV using a commercial probe labeled with digoxigenin
Keywords:
Cervix uteri., HPV DNA Probes., In situ hybridization., Penile.Abstract
Introduction: Infection with human papillomavirus (HPV) is considered the major risk factor for developing cancer and precancerous lesions of cervix, penis, anus, among others. Objective: To implement the chromogenic in situ hybridization (CISH) protocol with commercial probe for HPV of high risk (HR-HPV) (types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 69, 82) and of low risk (LR-HPV) (types 6, 11) in formalinfixed and paraffin-embedded samples. Material and method: CISH we re performed on biopsy samples with morphological changes suggestive of HPV, selected from the file of Pathology Center of Adolfo Lutz Institute. Thirty-eight cervical, 10 penile, 11 anal canal and five foreskin samples were analyzed for both HPV-LR and HR. Five different protocols were tested for the two probes, following modifications in the running time (t) and temperature (T) steps for hybridization, proteolysis, incubation with the detection system. Results: In current study, the increase of t and T parameters to proteolysis and incubation with the detection system, for the both probes, raised the labeling in the infected cells. LR-HPV probe was positive in 6.5 % of CIN 1 and in 60.0 % of penile with condyloma accuminata. HR-HPV probe was positive in 18.5 % of cervix and 7.4 % of penile samples. Two cases (one cervix and one penis) were positive for both probes. Conclusion: The implemented protocol proved to be suitable for the samples fixed in formalin and embedded in paraffin, for the public labora tory conditions
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Copyright (c) 2022 Lidia Midori Kimura Midori Kimura, Neuza Kasumi Shirata Kasumi Shirata, Suely Nonogaki Nonogaki, Juliana Mariotti Guerra, Marina Suheko Oyafuso, Yara de Menezes, Leonardo José Tadeu de Araújo, Celso di Loreto
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