The performance of the HPV16 real-time PCR integration assay

Ruutu, Merja; Kulmala, Satu-Maria; Peitsaro, Panu; Syrjänen, Stina

doi:10.1016/j.clinbiochem.2007.12.014

Cited by 23 publications

(23 citation statements)

References 16 publications

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“…In this regard, Ruutu et al state that an E2/E6 ratio ≥1 cannot entirely exclude the possibility of integration (40). However, the authors believe that this would be a rare phenomenon and emphasize the great sensitivity of the method.…”

Section: Discussionmentioning

confidence: 99%

“…Our real-time quantitative PCR (qPCR) strategy is based on a test described by Peitsaro et al, which has been extensively used for simultaneous detection of the viral load and physical state of the predominant HR-HPV type 16 (22, 28-34, 38, 39). Recent publications confirmed this test to be sensitive and specific for the identification of HPV integration (33,40). For this study, qPCR analysis has been applied on a large number of samples from a routine liquid-based cytology (LBC) setting to assess HPV16 viral load and integration state as risk markers for ≥CIN2 lesions in HPV16-positive women.…”

Section: Introductionmentioning

confidence: 99%

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Human Papillomavirus 16 Load and E2/E6 Ratio in HPV16-Positive Women: Biomarkers for Cervical Intraepithelial Neoplasia ≥2 in a Liquid-Based Cytology Setting?

Boulet

Benoy

Depuydt

et al. 2009

Cancer Epidemiology, Biomarkers &Amp; Prevention

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This retrospective case-control study assessed human papillomavirus 16 (HPV16) viral load and E2/E6 ratio as risk markers for cervical intraepithelial neoplasia (CIN) ≥2 lesions in HPV16-positive women in a routine liquid-based cytology setting. Triplex quantitative PCR for HPV16 E6, E2, and β-globin was done to determine the HPV16 load and the E2/E6 ratio, as a surrogate marker for integration, for women with a negative histologic endpoint (200 controls: 83 normal histology and 117 CIN1) and women with a ≥CIN2 endpoint (180 cases: 41 CIN2, 122 CIN3, and 17 invasive carcinoma). Our analysis showed a significantly higher HPV16 load in the case group, which was completely attributable to the high viral load of samples with invasive carcinoma as histologic endpoint. There was no significant difference in viral load between the other histologic groups. The E2/E6 ratio proved to be lower for the cases. However, the E2/E6 ratio indicated the presence of HPV integration in a considerable amount of control samples (44.3%), which suggests that HPV integration occurs early in the development of cancer and undermines the clinical value of viral integration. Overall, the intrinsic heterogeneous nature of the cervical cytology samples caused a substantial overlap of the HPV16 load and the E2/E6 ratio between controls and cases, which precludes the determination of cutoff values for risk prediction and hampers the clinical applicability in a cervical screening setting. (Cancer Epidemiol Biomarkers Prev 2009;18(11):2992-9)

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Human Papillomavirus 16 Load and E2/E6 Ratio in HPV16-Positive Women: Biomarkers for Cervical Intraepithelial Neoplasia ≥2 in a Liquid-Based Cytology Setting?

Boulet

Benoy

Depuydt

et al. 2009

Cancer Epidemiology, Biomarkers &Amp; Prevention

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“…However, the following three points should be taken into account: i) disrupted viral copies may be flanked by intact viral copies (40), ii) HPV integration does not always cause E2 disruption (41,42); and iii) more than a 10-fold excess of episomal form to integrated form of the virus interferes with E2 amplification, regardless the amount of viral DNA, resulting in lower E2/E6 ratios (43). Those factors tend to mask the presence of the integrated form HPV.…”

Section: ------------------------------------------------------------mentioning

confidence: 99%

Human papillomavirus is frequently detected in gefitinib-responsive lung adenocarcinomas

Baba

Castillo²,

Koriyama³

et al. 2010

Oncol Rep

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Abstract.A number of studies have reported the presence of human papillomavirus (HPV) in lung carcinoma. Interestingly, its detection rate appears to differ histologically and geographically. The present study examined 30 adenocarcinomas and 27 squamous cell carcinomas of the lung in a southern area of Japan, and detected high-risk HPV genome in 9 (30%) adenocarcinomas and 2 (7%) squamous cell carcinomas, using PCR with SPF10 primers and INNO-LiPA HPV genotyping assay. The difference of HPV detection rates in adenocarcinomas and squamous cell carcinomas was statistically significant (P=0.044, Fisher's exact test). HPV-16 was the most prevalent HPV genotype, and was detected in 27% (8/30) of adenocarcinomas and in 7% (2/27) of squamous cell carcinomas. High-risk-HPV positive carcinomas had decreased proportions of pRb (P=0.107) and significantly increased proportions of p16INK4a expressing cells (P=0.031) when compared to HPV-negative lung carcinomas. All HPV-16-positive cases were considered to have an integrated form of HPV-16 but its viral load was low (geometric mean = 0.02 copy per cell). In 20 additional adenocarcinomas treated with gefitinib, a tyrosine kinase inhibitor specific for epidermal growth factor receptor, the presence of HPV was examined. Note that East Asian ethnicity is a predictive factor of gefitinib response. High-risk HPV genome was found in 75% (6/8) of adenocarcinomas with complete or partial response to gefitinib but was not found in the remaining 12, which did not respond to gefitinib. In conclusion, the present study suggests that high-risk HPV may be more strongly related to adenocarcinomas, particularly gefitinib-responsive adenocarcinomas, when compared to squamous cell carcinomas. However, its low viral load makes it difficult to determine the etiological significance of these findings.

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“…On the other hand, integration of HPV DNA into the human genome is considered an essential step in the progression of HPV-associated cervical infection to invasive cervical carcinoma (18, 41). Upon integration the viral E2 gene, encoding a transcriptional repressor, is often disrupted with loss of its repressor function thus leading to up regulation of the transcription of the E6 and E7 oncogenes (30, 12). High expression of the viral oncogenes leads to disturbance of cell cycle controls and genomic instability that increase the risk of malignant transformation (25, 29).…”

Section: Introductionmentioning

confidence: 99%

Correlation between ebv co-infection and HPV16 genome integrity in Tunisian cervical cancer patients

Kahla¹,

Oueslati²,

Achour³

et al. 2012

Braz. J. Microbiol.

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Infection with high risk Human papillomavirus (HR-HPV) is necessary but not sufficient to cause cervical carcinoma. This study explored whether multiple HR-HPV or coinfection with Epstein-Barr virus (EBV) influence the integration status of HPV16 genome. The presence and typing of HPV in a series of 125 cervical specimens were assessed by polymerase chain reaction (PCR) using the specific primers for the HPV L1 region. As for EBV infection, the viral EBNA1 gene was used for its detection through PCR amplification. Disruption of the HPV E2 gene was assessed by amplification of the entire E2 gene with single set of primers, while E2 transcripts were evaluated by a reverse transcription PCR method (RT-PCR). The overall prevalence of HPVDNA was of 81.8% in cervical cancers versus 26.9% in benign lesions. In HPV positive cases, HPV16 and HPV18 were the most prevalent types, followed by HPV types 33, 31. EBV EBNA1 prevalence was statistically more frequent in cervical carcinomas than in benign lesions (29.5%, vs 9.6%; P=0.01). No viral infection was detected in healthy control women. The uninterrupted E2 gene was correlated with the presence of E2 transcripts originating from the HPV episomal forms. It was observed that integration was more common in HPV18 and EBV coinfection. The presence of EBV caused a five-fold [OR= 5; CI= 1.15-21.8; P = 0.04] increase in the risk of HPV16 genome integration in the host genome. This study indicates that EBV infection is acting as a cofactor for induction of cervical cancer by favoring HPVDNA integration.

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The performance of the HPV16 real-time PCR integration assay

Cited by 23 publications

References 16 publications

Human Papillomavirus 16 Load and E2/E6 Ratio in HPV16-Positive Women: Biomarkers for Cervical Intraepithelial Neoplasia ≥2 in a Liquid-Based Cytology Setting?

Human Papillomavirus 16 Load and E2/E6 Ratio in HPV16-Positive Women: Biomarkers for Cervical Intraepithelial Neoplasia ≥2 in a Liquid-Based Cytology Setting?

Human papillomavirus is frequently detected in gefitinib-responsive lung adenocarcinomas

Correlation between ebv co-infection and HPV16 genome integrity in Tunisian cervical cancer patients

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