Analysis of Human Papillomavirus Type 16 (HPV16) DNA Load and Physical State for Identification of HPV16-Infected Women with High-Grade Lesions or Cervical Carcinoma

Saunier, Maëlle; Monnier-Benoit, Sylvain; Mauny, Frédéric; Dalstein, Véronique; Briolat, Jenny; Riethmuller, Didier; Kantélip, B.; Schwarz, Elisabeth; Mougin, Christiane; Prétet, Jean‐Luc

doi:10.1128/jcm.01212-08

Cited by 94 publications

(107 citation statements)

References 54 publications

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“…Its protein product, E2, normally downregulates the expression of E6 and E7 (13)(14)(15). In the majority of HPV16-and HPV18-positive CCs, the virus has been detected in an integrated form (15)(16)(17)(18), although there are also HPV-positive CCs in which the virus is either in a mixed form or only in episomes (15,18).…”

Section: Introductionmentioning

confidence: 99%

Molecular Markers Implicating Early Malignant Events in Cervical Carcinogenesis

Koskimaa

Kurvinen

Costa

et al. 2010

Cancer Epidemiology, Biomarkers &Amp; Prevention

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Background: Human papillomavirus can induce a stepwise progression of precursor lesions to carcinoma. Sensitive and specific molecular markers are needed to identify the cervical lesions (CIN) at risk for this progression. hTERT activation could be one indicator of a point of no return in malignant progression.Methods: The UT-DEC-1 cell line is an in vitro model for the study of human papillomavirus-induced progression. Using molecular mining, nine potential genes interlinking hTERT and viral oncogene expression with the phenotypical features of CIN2 were identified. After preliminary testing with real-time PCR, five genes were selected for further analysis: hTERT, DKC1, Bcl-2, S100A8, and S100A9. These proteins were also tested in a series of 120 CIN lesions using immunohistochemistry.Results: Analysis of the mRNA expression of these genes at different cell passages revealed three time points with significant changes. hTERT, Bcl-2, and S100A9 were also overexpressed in CIN lesions, and the expression pattern changed during the progression toward CIN3 lesions.Conclusions: These identified time points that were combined with the mRNA overexpression of target genes matched events previously shown to be important in the progression toward malignancy: (a) the viral integration into the cell genome and episome loss; (b) the selection of cells with an acquired growth advantage and ability to maintain telomerase activity; and (c) the final stage of malignancy with permanently upregulated telomerase.Impact: hTERT, Bcl-2, and S100A9 together might compose a potential prognostic marker panel for the assessment of CIN lesions. These results, however, need further validation in prospective clinical settings.

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Section: Introductionmentioning

confidence: 99%

Molecular Markers Implicating Early Malignant Events in Cervical Carcinogenesis

Koskimaa

Kurvinen

Costa

et al. 2010

Cancer Epidemiology, Biomarkers &Amp; Prevention

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“…The significance of the viral load may vary with different types of HPV infection. For example, while HPV16 viral load was increased with the severity of lesions from precancerous lesion (squamous intraepithelial lesion) to invasive carcinoma, HPV18 viral load did not correlate with the severity of disease (Woodman et al, 2007;Saunier et al, 2008;Xi et al, 2009).…”

Section: Discussionmentioning

confidence: 98%

Prognostic Value of HPV18 DNA Viral Load in Patients with Early-Stage Neuroendocrine Carcinoma of the Uterine Cervix

Siriaunkgul

Utaipat²,

Suwiwat³

et al. 2012

Asian Pacific Journal of Cancer Prevention

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Objectives: To evaluate the clinicopathologic correlation and prognostic value of HPV18 DNA viral load in patients with early-stage cervical neuroendocrine carcinoma (NECA). Methods: Formalin-fixed, paraffinembedded tissue of cervical NECA patients with known HPV18 infection and clinicopathologic data including follow-up results were collected. The HPV18 DNA load was assessed with quantitative PCR targeting the HPV18 E6E7 region. Results: Twenty-one patients with early-stage (IB-IIA) cervical NECA were identified. HPV18 DNA viral load ranged from 0.83 to 55,174 copies/cell (median 5.90). Disease progression, observed in 10 cases (48%), was not significantly associated with any clinicopathologic variables. However, the group of patients with progressive disease tended to have a higher rate of pelvic lymph node metastasis (50% versus 9%, p=0.063) and a lower median value of HPV18 DNA viral load (4.37 versus 8.17 copies/cell, p=0.198) compared to the non-recurrent group. When stratified by a cut-off viral load value of 5.00 copies/cell, the group of patients with viral load ≤5.00 copies/cell had a significantly shorter disease-free survival than the group with viral load >5.00 copies/cell (p=0.028). The group with a lower viral load also tended to have a higher rate of disease progression (75% versus 31%, p=0.080). No significant difference in the other clinicopathologic variables between the lower and higher viral load groups was identified. Conclusion: HPV18 DNA viral load may have a prognostic value in patients with early-stage NECA of the cervix. A low viral load may be predictive of shortened disease-free survival in these patients.

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“…Diversi sono invece i lavori in letteratura che valutano l'influenza dell'arricchimento del pellet mediante centrifugazione dei tamponi cervicali per la citologia su strato sottile (ThinPrep, SurePath) per aumentare la sensibilità del test molecolare rispetto all'estrazione diretta del DNA (5,12,35), mentre praticamente inesistenti sono lavori che valutino l'idoneità del prelievo eseguito con i tamponi E-Swab (Copan, Italia), i più utilizzati attualmente nelle microbiologie perché adatti all'esecuzione automatizzata degli esami colturali. Inesistenti sono anche lavori condotti con metodiche NAATs che quantificano la carica definendo dei cut-off per il trattamento dei patogeni commensali opportunisti (Ureaplasma spp.…”

Section: Introduzioneunclassified

Pre-analytical and post-analytical evaluation in the era of molecular diagnosis of sexually transmitted diseases: cellularity control and internal control

Bianchi¹,

Napoli²,

Turrisi³

et al. 2014

Microbiol Med

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SummaryPre-analytical and post-analytical evaluation in the era of molecular diagnosis of sexually transmitted diseases: cellularity control and internal control.Background. Increase of molecular tests performed on DNA extracted from various biological materials should not be carried out without an adequate standardization of the pre-analytical and postanalytical phase.Materials and Methods. Aim of this study was to evaluate the role of internal control (IC) to standardize pre-analytical phase and the role of cellularity control (CC) in the suitability evaluation of biological matrices, and their influence on false negative results. 120 cervical swabs (CS) were pre-treated and extracted following 3 different protocols. Extraction performance was evaluated by amplification of: IC, added in each mix extraction; human gene HPRT1 (CC) with RT-PCR to quantify sample cellularity; L1 region of HPV with SPF10 primers. 135 urine, 135 urethral swabs, 553 CS and 332 ThinPrep swabs (TP) were tested for C. trachomatis (CT) and U. parvum (UP) with RT-PCR and for HPV by endpoint-PCR. Samples were also tested for cellularity.Results. Extraction protocol with highest average cellularity (Ac)/sample showed lowest number of samples with inhibitors; highest HPV positivity was achieved by protocol with greatest Ac/PCR. CS and TP under 300.000 cells/sample showed a significant decrease of UP (P<0.01) and HPV (P<0.005) positivity. Female urine under 40.000 cells/mL were inadequate to detect UP (P<0.05).Conclusions. Our data show that IC and CC allow optimization of pre-analytical phase, with an increase of analytical quality. Cellularity/sample allows better sample adequacy evaluation, crucial to avoid false negative results, while cellularity/PCR allows better optimization of PCR amplification. Further data are required to define the optimal cut-off for result normalization.

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Analysis of Human Papillomavirus Type 16 (HPV16) DNA Load and Physical State for Identification of HPV16-Infected Women with High-Grade Lesions or Cervical Carcinoma

Cited by 94 publications

References 54 publications

Molecular Markers Implicating Early Malignant Events in Cervical Carcinogenesis

Molecular Markers Implicating Early Malignant Events in Cervical Carcinogenesis

Prognostic Value of HPV18 DNA Viral Load in Patients with Early-Stage Neuroendocrine Carcinoma of the Uterine Cervix

Pre-analytical and post-analytical evaluation in the era of molecular diagnosis of sexually transmitted diseases: cellularity control and internal control

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