Standardization of the chromogenic in situ hybridization protocol for searching the high and low risk HPV using a commercial probe labeled with digoxigenin
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Keywords

Cervix uteri.
HPV DNA Probes.
In situ hybridization.
Penile.

How to Cite

1.
Midori Kimura LMK, Kasumi Shirata NKS, Nonogaki SN, Mariotti Guerra J, Suheko Oyafuso M, de Menezes Y, José Tadeu de Araújo L, di Loreto C. Standardization of the chromogenic in situ hybridization protocol for searching the high and low risk HPV using a commercial probe labeled with digoxigenin. Bepa [Internet]. 2022 Aug. 7 [cited 2024 Nov. 22];14(158). Available from: https://periodicos.saude.sp.gov.br/BEPA182/article/view/37992

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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