Detection of HPV-DNA by a PCR-based Method in Formalin-fixed, Paraffin-embedded Tissue From Rare Endocervical Carcinoma Types

Context.— Human papillomavirus (HPV) DNA in situ hybridization (ISH) assay and p16 immunohistochemistry (IHC) are used to determine high-risk HPV status in formalin-fixed, paraffin-embedded (FFPE) tissues in oropharyngeal squamous cell carcinoma (SCC). Although high sensitivity and specificity for HPV can be obtained by combined p16 IHC and HPV DNA ISH, the occasional discrepancy between these assays has prompted evaluation of Cervista HPV assays in FFPE tissue from patients with oropharyngeal SCC. Objective.— To compare the efficacy of Cervista HPV 16/18 and Cervista HPV HR assay to that of HPV DNA ISH assay and p16 IHC in FFPE tissue in head and neck squamous cell carcinoma of oropharyngeal origin. Design.— Archived FFPE tissue from 84 patients with SCC of oropharyngeal origin and available HPV DNA ISH and p16 IHC test results were tested with the Cervista HPV 16/18 assay and further verified by polymerase chain reaction (PCR)–based HPV16/18 genotyping tests in cases with discrepancy. Results.— Of the 84 specimens, 75% (63 of 84) were positive and 16% (13 of 84) had discrepant or equivocal findings by p16 IHC and HPV DNA ISH testing. Use of Cervista HPV assays, either to clarify discrepant/equivocal findings or as confirmation after initial p16 IHC/HPV DNA ISH tests, identified 81% (68 of 84) of HPV-positive cases without equivocal HPV results. Five of 13 cases with discrepancy or equivocal HPV DNA ISH results tested positive for HPV16 or HPV18 by Cervista HPV 16/18 assay, which was further confirmed by PCR-based HPV 16/18 genotyping. Conclusions.— The Cervista HPV assays are a reasonable alternative to HPV DNA ISH in determining HPV status in FFPE tissue specimens from patients with oropharyngeal SCC.

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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 ◽

10.1155/2013/453606 ◽

2013 ◽

Vol 2013 ◽

pp. 1-11 ◽

Cited By ~ 11

Author(s):

Roberta Zappacosta ◽

Antonella Colasante ◽

Patrizia Viola ◽

Tommaso D’Antuono ◽

Giuseppe Lattanzio ◽

...

Keyword(s):

In Situ Hybridization ◽

Dna Test ◽

Hpv Dna ◽

Formalin Fixed Paraffin ◽

Formalin Fixed Paraffin Embedded ◽

Chromogenic In Situ Hybridization ◽

High Level ◽

Formalin Fixed ◽

Hpv Dna Test

Although HPV-DNA test and E6/E7 mRNA analyses remain the current standard for the confirmation of human papillomavirus (HPV) infections in cytological specimens, no universally adopted techniques exist for the detection of HPV in formalin-fixed paraffin-embedded samples. Particularly, in routine laboratories, molecular assays are still time-consuming and would require a high level of expertise. In this study, we investigated the possible use of a novel HPV tyramide-based chromogenic in situ hybridization (CISH) technology to locate HPV on tissue specimens. Then, we evaluate the potential usefulness ofp16INK4a/Ki-67 double stain on histological samples, to identify cervical cells expressing HPV E6/E7 oncogenes. In our series, CISH showed a clear signal in 95.2% of the specimens and reached a sensitivity of 86.5%. CISH positivity always matched with HPV-DNA positivity, while 100% of cases with punctated signal joined with cervical intraepithelial neoplasia grade 2 or worse (CIN2+). p16/Ki67 immunohistochemistry gave an interpretable result in 100% of the cases. The use of dual stain significantly increased the agreement between pathologists, which reached 100%. Concordance between dual stain and E6/E7 mRNA test was 89%. In our series, both CISH andp16INK4a/Ki67 dual stain demonstrated high grade of performances. In particular, CISH would help to distinguish episomal from integrated HPV, in order to allow conclusions regarding the prognosis of the lesion, whilep16INK4a/Ki67 dual stain approach would confer a high level of standardization to the diagnostic procedure.

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Evaluation of two commercialised in situ hybridisation assays for detecting HPV-DNA in formalin-fixed, paraffin-embedded tissue

Archives of Gynecology and Obstetrics ◽

10.1007/s00404-010-1771-z ◽

2010 ◽

Vol 284 (4) ◽

pp. 999-1005 ◽

Cited By ~ 3

Author(s):

Maresa Montag ◽

Thomas J. F. Blankenstein ◽

Naim Shabani ◽

Ansgar Brüning ◽

Ioannis Mylonas

Keyword(s):

In Situ Hybridisation ◽

Hpv Dna ◽

Formalin Fixed Paraffin ◽

Formalin Fixed Paraffin Embedded ◽

Formalin Fixed

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Evaluation of the Anyplex II HPV28 Assay in the Detection of Human Papillomavirus in Archival Samples of Oropharyngeal Carcinomas

Archives of Pathology & Laboratory Medicine ◽

10.5858/arpa.2019-0199-oa ◽

2019 ◽

Vol 144 (5) ◽

pp. 620-625 ◽

Cited By ~ 2

Author(s):

Francesca Rollo ◽

Maria Gabriella Donà ◽

Barbara Pichi ◽

Raul Pellini ◽

Renato Covello ◽

...

Keyword(s):

Human Papillomavirus ◽

Detection Rate ◽

The Other ◽

Hpv Dna ◽

Formalin Fixed Paraffin ◽

Hpv Status ◽

Formalin Fixed Paraffin Embedded ◽

Good Concordance ◽

Cancer Tissues ◽

Formalin Fixed

Context.— The improved survival and better response to treatments of human papillomavirus (HPV)–related oropharyngeal squamous cell carcinoma (OPSCC) highlight the need for effective tools in evaluating HPV status on formalin-fixed, paraffin-embedded (FFPE) cancer tissues. To date, there is no agreement regarding the most appropriate method for HPV testing on FFPE materials. Objective.— We aimed to investigate the performance of the Anyplex II HPV28 (Anyplex) on FFPE OPSCC tissues and to compare it with 2 other methods for HPV-DNA detection and p16 overexpression. Design.— One hundred sixty FFPE OPSCCs were evaluated, which had already been analyzed with the INNO-LiPA HPV assay, Xpert HPV assay, and p16 immunostaining. Results.— All the samples but 1 provided valid results with the Anyplex, which showed the highest HPV detection rate and a good concordance with all the other methods (κ = 0.75, 95% CI, 0.65–0.85 versus INNO-LiPA; κ = 0.80, 95% CI, 0.70–0.89 versus Xpert; κ = 0.76, 95% CI, 0.65–0.86 versus p16). Moreover, the HPV-driven fraction, based on HPV-DNA and p16 double positivity, was higher with Anyplex (83 of 159, 52.2%) than with the other 2 assays, that is, 78 of 156 (50.0%) for INNO-LiPA and 80 of 160 (50.0%) for Xpert. Conclusions.— Anyplex II HPV28 showed a higher HPV detection rate and HPV-associated fraction than the other methods used. This assay is suitable for HPV detection in archival OPSCC tissues.

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Immunohistochemical Detection of early and late Human Papillomaviruses (HPV) proteins in Retinoblastoma tumor

10.1101/2020.07.16.20153882 ◽

2020 ◽

Author(s):

Nara Diniz Soares Pessoa ◽

Thatiana Correia Melo ◽

Rodrigo Pinheiro Araldi ◽

Rofrigo Franco Carvalho ◽

Willy Becak ◽

...

Keyword(s):

Hpv Infection ◽

Human Papillomaviruses ◽

Immunohistochemical Detection ◽

Hpv Dna ◽

Formalin Fixed Paraffin ◽

Formalin Fixed Paraffin Embedded ◽

Retinoblastoma Tumor ◽

Formalin Fixed

In this study, we evaluated the presence of early and late Human Papillomavirus (HPV) proteins in retinoblastoma Brazilian patients. For this, 8 formalin-fixed paraffin-embedded retinoblastoma tissue blocks were used. HPV DNA presence was determined by in situ hybridization (ISH). Immunohistochemistry were performed to verify HPV16/18 E6, E1^E4, and L1 proteins. HPV was detected in all retinoblastoma tumors and viral DNA was labeled in tumor cells, retinal layers and optical nerve structures. In addition, E1^E4, E6 and L1 proteins were detected in all samples in the same areas where HPV DNA was detected. Our data showed the presence and expression of early and late HPV proteins in retinoblastoma tumors from Brazilian children. However, further studies should be performed to clarify the role of HPV infection in retinoblastoma tumor.

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