Detection of human papillomavirus DNA in CaSki and HeLa cells by fluorescent in situ hybridization

Lizard, Gérard; Chignol, Marie‐Christine; Chardonnet, Y; Souchier, Catherine; Bordes, M.; Schmitt, Daniel; Revillard, Jean‐Pierre

doi:10.1016/0022-1759(93)90067-h

Cited by 22 publications

(15 citation statements)

References 23 publications

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“…In the cervical cancer cell lines, viral load values of 1.5 copies/cell HPV 16 in SiHa, 548 copies/cell HPV 16 in CaSki, and 8 copies/cell HPV 18 in HeLa were obtained which were in agreement with published results obtained using other methodologies (Ho et al, 1995; Lizard et al, 1993; Syrjanen et al, 1991). Linear results were observed in both the uniplex and multiplex platforms on serial 10-fold dilutions of HeLa DNA (1.37 pg/μl - 13.7 ng/μl), with calculated HPV 18 concentrations decreasing proportionally as input sample DNA concentrations decreased.…”

Section: Resultssupporting

confidence: 89%

Development and evaluation of a PCR and mass spectroscopy (PCR–MS)-based method for quantitative, type-specific detection of human papillomavirus

Patel

Shih

Newton

et al. 2009

Journal of Virological Methods

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Knowledge of the central role of high-risk human papillomavirus (HPV) in cervical carcinogenesis, coupled with an emerging need to monitor the efficacy of newly introduced HPV vaccines, warrant development and evaluation of type-specific, quantitative HPV detection methods. In the present study, a prototype PCR and mass spectroscopy (PCR-MS)-based method to detect and quantitate 13 high-risk HPV types is compared to the Hybrid Capture 2 High Risk HPV DNA test (HC2; Digene Corp., Gaithersburg, MD) in 199 cervical scraping samples and to DNA sequencing in 77 cervical tumor samples. High-risk HPV types were detected in 76/77 (98.7%) cervical tumor samples by PCR-MS. Degenerate and type-specific sequencing confirmed the types detected by PCR-MS. In 199 cervical scraping samples, all 13 HPV types were detected by PCR-MS. Eighteen (14.5%) of 124 cervical scraping samples that were positive for high-risk HPV by HC2 were negative by PCR-MS. In all these cases, degenerate DNA sequencing failed to detect any of the 13 high-risk HPV types. Nearly half (46.7%) of the 75 cervical scraping samples that were negative for high-risk HPV by the HC2 assay were positive by PCR-MS. Type-specific sequencing in a subset of these samples confirmed the HPV type detected by PCR-MS. Quantitative PCR-MS results demonstrated that 11/75 (14.7%) samples contained as much HPV copies/cell as HC2-positive samples. These findings suggest that this prototype PCR-MS assay performs at least as well as HC2 for HPV detection, while offering the additional, unique advantages of type-specific identification and quantitation. Further validation work is underway to define clinically meaningful HPV detection thresholds and to evaluate the potential clinical application of future generations of the PCR-MS assay.

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Section: Resultssupporting

confidence: 89%

Development and evaluation of a PCR and mass spectroscopy (PCR–MS)-based method for quantitative, type-specific detection of human papillomavirus

Patel

Shih

Newton

et al. 2009

Journal of Virological Methods

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“…Human cervical cancer cell lines CaSki (contain about 600 copies of HPV 16), HeLa (contain 20-50 copies of HPV 18) (Lizard et al, 1993) and the human mammary glands epithelial cell line HBL-100, all obtained from the ATCC (Manassas, VA, USA), were resuscitated by the Cell Bank of Shanghai Institute of Biochemistry and Cell Biology, CAS (Shanghai, PR China). Cells were cultured according to the ATCC's specifications.…”

Section: Cell Culture and Reagentsmentioning

confidence: 99%

HPV E6 down-regulation and apoptosis induction of human cervical cancer cells by a novel lipid-soluble extract (PE) from Pinellia pedatisecta Schott in vitro

Jiang

Xia

et al. 2010

Journal of Ethnopharmacology

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“…These findings support previous studies confirming the theory that p16 INK4A is overexpressed in dysplastic cells of the cervix 47 INK4A and HPV copy number is typical of the response of a cell to the action of an agonist 56 and in figure 2, is approximated by a fit with the following function y=a+b√x 10 +cx, where a=62.4, b=3.5 and c=-0.1. It should be noted that, in figure 2, the HPV copy number is represented by the average of the range quoted in literature [57][58] and so error margins in the horizontal axis are potentially very large. However, the sublinear nature of the plot indicates that p16 INK4A expression levels are 15 particularly sensitive for low HPV copy number.…”

Section: Confocal Microscopymentioning

confidence: 99%

Correlation of p16INK4A expression and HPV copy number with cellular FTIR spectroscopic signatures of cervical cancer cells

Ostrowska¹,

Garcia²,

Meade³

et al. 2011

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Detection of human papillomavirus DNA in CaSki and HeLa cells by fluorescent in situ hybridization

Cited by 22 publications

References 23 publications

Development and evaluation of a PCR and mass spectroscopy (PCR–MS)-based method for quantitative, type-specific detection of human papillomavirus

Development and evaluation of a PCR and mass spectroscopy (PCR–MS)-based method for quantitative, type-specific detection of human papillomavirus

HPV E6 down-regulation and apoptosis induction of human cervical cancer cells by a novel lipid-soluble extract (PE) from Pinellia pedatisecta Schott in vitro

Correlation of p16INK4A expression and HPV copy number with cellular FTIR spectroscopic signatures of cervical cancer cells

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