HPV DNA Detection by In Situ Hybridization with Catalyzed Signal Amplification on Thin-layer Cervical Smears

Samama, Brigitte; Plas-Roser, S.; Schaeffer, C.; Château, Danielle; Fabre, M; Boehm, Nelly

doi:10.1177/002215540205001014

Cited by 25 publications

(25 citation statements)

References 14 publications

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“…A diffuse hybridization pattern was not observed. Others have shown that the punctate pattern of hybridization correlates with viral DNA integration, whereas a diffuse signal correlates with episomal HPV (31). The hybridization signals were strictly limited to the carcinoma and the associated dysplastic epithelium.…”

Section: Resultsmentioning

confidence: 88%

Tissue Distribution of Human Papillomavirus 16 DNA Integration in Patients with Tonsillar Carcinoma

Begum¹,

Cao²,

Gillison³

et al. 2005

Clinical Cancer Research

295

215

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Purpose: Human papillomavirus16 (HPV-16) has been implicated as a causative agent in a subset of head and neck squamous cell carcinomas (HNSCC). This study was undertaken to discern the distribution and timing of HPV viral integration during tumorigenesis of the upper respiratory tract. Experimental Design: A tissue array was assembled from a consecutive group of 176 patients with HNSCCs. The array was evaluated by HPV-16 in situ hybridization and p16 immunohistochemistry. Patients with HPV-positive tonsillar cancers who had undergone bilateral tonsillectomies were selected for more complete mapping of viral integration. Results: HPV-16 was detected in 38 of the 176 (22%) cases by in situ hybridization. When stratified by site of origin, HPV-16 was detected in 37 of 45 cancers arising from the oropharynx but in only 1of 131tumors arising from nonoropharyngeal sites (82% versus 0.8%, P < 0.00001).P16 expression was associated with the presence of HPV-16: 31 of 38 HPV-positive tumors exhibited p16 expression, whereas only 9 of the 138 HPV-negative tumors were p16-positive (82% versus 6%, P < 0.00001). In the bilateral tonsil sections, hybridization signals were strictly limited to the invasive cancers and associated dysplasias. P16 staining was widely distributed throughout the nonneoplastic crypt epithelium of individuals with and without tonsillar cancer. Conclusions: HPV-16 is strongly associated with carcinomas arising from the oropharynx, and integration is tightly coupled to the neoplastic process.Viral integration does not occur as a field alteration throughout normal tonsillar epithelium. P16 expression localizes to HPV-positive cancers, and is intrinsic to the specialized epithelium of the tonsillar crypts. For risk assessment, early cancer detection and disease surveillance, evidence of HPV-16 integration may represent a meaningful finding, whereas high p16 expression, by itself, may not.

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Section: Resultsmentioning

confidence: 88%

Tissue Distribution of Human Papillomavirus 16 DNA Integration in Patients with Tonsillar Carcinoma

Begum¹,

Cao²,

Gillison³

et al. 2005

Clinical Cancer Research

295

215

View full text Add to dashboard Cite

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“…This method allowed us to detect as few as one or two copies of HPV DNA as previously described [9]. The physical state of the virus (integrated, episomal, or mixed) had been determined by the pattern of the ISH signal, a dotted pattern corresponding to the integrated form of the virus, a diffuse pattern to its episomal form [9,11,12].…”

Section: Hpv Detection By Ishmentioning

confidence: 99%

“…In situ hybridization was performed by using cold probes (Enzo Diagnostic, New York, NY) revealing the 6/11, 16/18, and 31/33/51 HPV genomes according to a procedure previously described [9]. Briefly, the slides were deparaffinized with toluene, rehydrated with absolute alcohol, and finally rinsed with water.…”

Section: Hpv Detection By Ishmentioning

confidence: 99%

“…Therefore, we examined the expression of p16 in relation to the physical status of HPV (episomal or integrated) in a series of 110 formalin-fixed, paraffin-embedded cutaneomucous biopsies for HPV screening and typing, as the hematoxylin-eosin (HE) staining was suggestive of an HPV infection. Human papillomavirus 6, 11, 16, 18, 31, 33, and 51 were detected using in situ hybridization (ISH) [9] with tyramide amplification and p16 expression was screened by immunohistochemistry. Although some reports have described the relationship between the presence of high-risk HPV and p16 overexpression in anogenital lesions [4,10], to our knowledge, this is the first study analyzing the correlation between p16 expression and HPV physical status in a large series of anogenital lesions.…”

Section: Introductionmentioning

confidence: 99%

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p16 expression in relation to human papillomavirus in anogenital lesions

2006

Self Cite

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“…Subsequent adaptation of the method to slide-based applications has demonstrated remarkable gains in sensitivity for FISH for detection of protein (Davies et al 1999), DNA (Acar et al 2002;McKay et al 1997;Schriml et al 1999), and RNA targets (Breininger and Baskin 2000;Qian et al 2001;Tata 2001;van de Corput et al 1998a, b;van Gijlswijk et al 2000;Wagner et al 1998;Yang et al 1999;Zaidi et al 2000;Zhang and Kitching 2000), and for bright field molecular applications (Zehbe et al 1997;Tubbs et al 2002a, b;Speel et al 1998;Samama et al 2002;Wiedorn et al 1999;Plummer et al 1998;Sano et al 1998). In the present study we combined the two approaches for simultaneous and robust immunophenotypic segregation of populations for genotyping using multicolored FISH probes.…”

Section: Introductionmentioning

confidence: 99%

Genotyping of phenotypically defined cells in neoplasia: enhanced immunoFISH via tyramide signal amplification (TSA) segregates immunophenotypically—defined cell populations for gated genotyping

et al. 2007

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Molecular morphologic tools exist for simultaneously visualizing immunophenotype and genotype of tumors, but are frequently hampered by a delicate balance between removing sufficient amount of the protein blocking full access of the probe to hybridize to target nucleic acids while still preserving sufficient target antigen for immunophenotyping. The result is often suboptimal, with either insufficiently visualized gene deletions and amplifications due to masking protein, or overdigestion of the protein target. Our purpose was to design and validate a gated genotyping assay that enables optimal and concomitant detection of both gene and protein. Using the proliferating endothelial cell compartment within gliomas organized in a tissue microarray (TMA), we tested the hypothesis that tyramide signal amplification (TSA) with deposition of a fluorochrome could be used during immunophenotyping, permitting sufficient protein digestion while insuring probe accessibility to nucleic acid target. The method was successfully validated using a TMA containing 38 glioma cases previously genotyped for EGFR amplification. CD31 positive endothelial cells were segregated via TSA-based Alexa-Fluor 647 immunofluorescence for analysis of EGFR amplification of the gliomas organized in the TMA. Enhanced immunoFISH (TSA) successfully segregates immunophenotypically-defined cell populations for gated genotyping.

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HPV DNA Detection by In Situ Hybridization with Catalyzed Signal Amplification on Thin-layer Cervical Smears

Cited by 25 publications

References 14 publications

Tissue Distribution of Human Papillomavirus 16 DNA Integration in Patients with Tonsillar Carcinoma

Tissue Distribution of Human Papillomavirus 16 DNA Integration in Patients with Tonsillar Carcinoma

p16 expression in relation to human papillomavirus in anogenital lesions

Genotyping of phenotypically defined cells in neoplasia: enhanced immunoFISH via tyramide signal amplification (TSA) segregates immunophenotypically—defined cell populations for gated genotyping

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