Evaluation of quantity and staining pattern of human papillomavirus (HPV)-infected epithelial cells in thin-layer cervical specimens using optimized HPV-CARD assay

Algeciras‐Schimnich, Alicia; Policht, Frank; Sitailo, Svetlana; Song, Meng; Morrison, Larry E.; Соколова, И. А.

doi:10.1002/cncr.22946

Cited by 14 publications

(19 citation statements)

References 55 publications

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“…Integration of the HPV DNA was found in some HSIL and CA lesions, but not in any patients with the cytology results of LSIL or NILM, suggesting that the event of integration may begin at some stage of LSIL. A recent paper described observing the S pattern using their system in almost half of the HSILs and even in some patients of ASCUS or LSIL [51]. The sensitivity of their ISH was similar to that of PCR.…”

Section: Discussionmentioning

confidence: 98%

“…The slides were incubated with 1.25 μg/ml proteinase K solution for 10 min at room temperature, washed with water, and then quenched with 0.3% H 2 O 2 in methanol for 20 min at room temperature before being finally washed again with water. Excess liquid was removed by air drying, and one drop of the high-risk HPV probe, consisting of a cocktail directed against the high-risk HPV types 16,18,31,33,35,39,45,51,52,56,58,59, and 68 (Dako Norden A/S), was applied. The slides were coverslipped.…”

Section: In Situ Hybridizationmentioning

confidence: 99%

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Ancillary testing of liquid-based cytology specimens for identification of patients at high risk of cervical cancer

et al. 2008

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Integration of human papillomavirus DNAs into the host genome is crucial to the development of cervical cancer. Overexpression of the P16 protein has been reported in cervical intraepithelial neoplasia (CIN) as well as cervical cancer. Such molecular biomarkers have been utilized for ancillary testing of liquid-based cytology specimens; however, their clinical application remains controversial. To detect CIN 2 or more advanced lesions, 153 liquid-based cytology (LBC) specimens were investigated to determine the physical status of the human papillomavirus (HPV) DNA by in situ hybridization (ISH) and to detect overexpression of the P16 protein by immunocytochemistry combined with HPV genotyping by polymerase chain reaction. The combination of ISH, P16 immunocytochemistry, and LBC showed high sensitivity (89.3%) as well as high specificity (92.6%). We confirmed the usefulness of P16 immunocytochemistry combined with ISH and HPV genotyping as ancillary molecular-biological tests of LBC specimens for identifying patients at high risk of cervical cancer.

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Section: Discussionmentioning

confidence: 98%

Section: In Situ Hybridizationmentioning

confidence: 99%

Ancillary testing of liquid-based cytology specimens for identification of patients at high risk of cervical cancer

et al. 2008

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“…We referred to as a multipunctate (MP) pattern. This second pattern has been correlated with viral integration [7,8]. Nuclear morphology was visible in the MP pattern but not in the DI pattern.…”

Section: Sample Proceduresmentioning

confidence: 90%

Detection of HPV by in situ hybridization in thin-layer (ThinPrep) cervicovaginal samples

et al. 2011

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We have studied an automated in situ hybridization (ISH) method as a possible alternative approach for detecting high-risk human papillomavirus (HPV) in monolayer (ThinPrep) cervico-vaginal samples, comparing the results with those obtained by polymerase chain reaction (PCR) using consensus primers and studying the relationship between the ISH staining pattern and the viral integration in HPV 16-positive cases. Eighty atypical squamous cells of undetermined significance (ASCUS) and low-grade squamous intraepithelial lesion (LSIL) cases were used for our purposes. The patients were monitored through periodic cytologies. ISH with was performed with an automated Ventana System, analysis by PCR was performed with consensus primers and integration of HPV16 was performed by realtime PCR analyzing E2 and E6 genes. Additionally, 27 HSIL cases were also studied to observe the ISH staining patterns. HPV infection was detected by ISH in 21.7% of the ASCUS cases and 55.8% of the LSIL cases. Two distinct staining patterns were observed: multipunctated (MP) and diffuse (DI). In some cases, a mixed pattern (MP + DI) was observed and these cases were considered as MP. The MP pattern increased with the degree of lesion and seemed to have a prognostic value in ASCUS/LSIL cases. The lesion in MP pattern cases persisted throughout the entire study in 77% of cases, whereas in cases with a DI staining pattern, only 41% of them showed persistence of the lesion (p <0.001). No correlation was found between HPV integration and the ISH staining pattern. Given the lower sensitivity and negative predictive value of ISH and its incapacity to demonstrate the integration of high-risk HPV in ASCUS and LSIL cases using liquid-based cytology, we do not recommend this technique for the triage of ASCUS and LSIL cases.

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“…HPV staining was visualized with a green filter and localized to the nucleus as confirmed by dAPI staining. HPV staining pattern was classified as diffuse, mixed and punctate ( Figure 1) as described previously [12,13]. The sample was considered positive for HPV infection if at least one HPV-positive cell was found.…”

Section: Methodsmentioning

confidence: 99%

“…Particularly, triple-color Vysis Cervical FISH Probe Kit was designed to identify HPV infected cells and determine copy number changes of the hTERC and MYC gene via FISH (HPV-FISH) [12,13].…”

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confidence: 99%

Molecular cytogenetic analyses of hTERC (3q26) and MYC (8q24) genes amplifications in correlation with oncogenic human papillomavirus infection in Czech patients with cervical intraepithelial neoplasia and cervical carcinomas

Kuglík¹,

Kašíková²,

Smetana³

et al. 2015

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It is known that cervical cancer develop from precancerous intraepithelial neoplasia (CIN) which is characterized by series of genetic abnormalities. The progression of CIN to cervical carcinoma has been associated especially with the genomic integration of oncogenic human papilloma virus (HPV) and gain of the human telomerase RNA gene hTERC (3q26) and MYC (8q24).In this study, cytology specimens of cervical intraepithelial neoplasia and cervical carcinoma from 74 Czech women were analyzed using the triple-color Cervical FISH Probe Kit designed for identification of HPV infected cells and copy number aberration of the hTERC and MYC genes. HPV-positivity exhibited 70% of patients with premalignant lesions (CIN I -CIN III, carcinoma in situ), chromosomal changes were found in 53.3% of cases -MYC amplification had 33.3% of women with CIN I -CIN III and 50% with carcinoma in situ. Amplification of hTERC was detected in 16.7% of patient with CIN I, in 50% with CIN II, in 58.3% with CIN III and in 66.7% with carcinoma in situ. Based on HPV-positivity and the occurrence of chromosomal aberrations, patients were divided into high-, intermediate-and low-risk group.Among women with cervical carcinomas, HPV infection was detected in 90.1% of specimens and chromosomal aberrations were found in 87.5% of samples. Amplification of MYC gene was detected in 25% and hTERC gene in 62.5% of patients. According to the histopathological grade of tumors, MYC gene amplification occurred more frequently in specimens of spinocellular carcinoma than adenocarcinoma (p=0.029). We found no association between the frequency of cytogenetic lesions and the incidence of lymphangiogenesis or lymph node metastases in cervical carcinoma patients. Simultaneous hTERC and MYC genes amplification was significantly more frequent in samples of cervical carcinomas than in premalignant lesions (p=0.008).In a cohort of 26 patients with cervical carcinoma we used oligo-based GGH+SNP microarray technique for the high resolution mapping of copy number changes of hTERC and MYC genes. We found that recurrent gain of genetic material in chromosome 3q26 area carrying hTERC gene of size 43.6 Mb between 3q25.1-3qter and duplication of 3q were the most common genomic identifications of amplified gene. In MYC locus array-CGH profiling identified duplication of 8q and trisomy 8 as frequent genomic changes.Our work confirmed that in cervical carcinoma gains of hTERC and MYC genes are specific genomic changes associated with developing of malignant phenotype. We also showed that in premalignant stages HPV-FISH assay can be used as an effective diagnostic procedure to identify patients carrying highly risking HPV infection and chromosomal aberrations associated with this malignancy.

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Evaluation of quantity and staining pattern of human papillomavirus (HPV)-infected epithelial cells in thin-layer cervical specimens using optimized HPV-CARD assay

Cited by 14 publications

References 55 publications

Ancillary testing of liquid-based cytology specimens for identification of patients at high risk of cervical cancer

Ancillary testing of liquid-based cytology specimens for identification of patients at high risk of cervical cancer

Detection of HPV by in situ hybridization in thin-layer (ThinPrep) cervicovaginal samples

Molecular cytogenetic analyses of hTERC (3q26) and MYC (8q24) genes amplifications in correlation with oncogenic human papillomavirus infection in Czech patients with cervical intraepithelial neoplasia and cervical carcinomas

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