Rapid and Sensitive Detection of Physical Status of Human Papillomavirus Type 16 DNA by Quantitative Real-Time PCR

Nagao, Shoji; Yoshinouchi, Mitsuo; Miyagi, Yasunari; Hongo, Atsushi; Kodama, Junichi; Itoh, Sachio; Kudo, Takafumi

doi:10.1128/jcm.40.3.863-867.2002

Cited by 84 publications

(77 citation statements)

References 23 publications

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“…In contrast to advanced lesions and cervical cancer (36), integration of HPV DNA into the host genome has been considered a rare event in early preneoplastic lesions of the cervix. Recent studies that used novel quantitative real-time PCR techniques, however, have demonstrated that integration of HPV may occur in early CIN lesions (37) and even in those containing HPV but no CIN (38). Also, rapid progression, in 1 to 2 years, from non-CIN lesions or CIN 2 to CIN 3 was shown to be associated with a heavy load of integrated HPV.…”

Section: Discussionmentioning

confidence: 99%

Folate Is Associated with the Natural History of High-Risk Human Papillomaviruses

Piyathilake

Henao²,

Macaluso³

et al. 2004

Cancer Research

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Several micronutrients have been implicated in cervical carcinogenesis. However, their mode of action is still a matter of speculation. In particular, it is unclear whether certain nutrients reduce the probability of acquiring high-risk human papillomavirus (HPV) or whether they facilitate the clearance of high-risk HPV. We conducted a 24-month prospective follow-up study to test the hypothesis that systemic concentrations of folate are associated with the occurrence and duration of high-risk HPV infections after controlling for other micronutrients (vitamins B 12 , A, E, and C, total carotene) and known risk factors for high-risk HPV infections and cervical cancer. Circulating concentrations of these micronutrients and risk factors for cervical cancer were determined in a cohort of 345 women who were at risk of developing cervical intraepithelial neoplasia. Using the hybrid capture 2 (HC-2) assay, high-risk HPV status was evaluated at 6-month intervals up to 24 months. All women had at least three consecutive visit high-risk HPV test results. Higher folate status was inversely associated with becoming HC-2 test-positive [odds ratio (OR): 0.27; 95% confidence interval (CI), 0.08 -0.91; P ‫؍‬ 0.04]. Women with higher folate status were significantly less likely to be repeatedly HC-2 test-positive (OR: 0.33; 95% CI, 0.13-0.86; P ‫؍‬ 0.02) and more likely to become test-negative during the study (OR: 2.50; 95% CI, 1.18 -5.30; P ‫؍‬ 0.02). To our knowledge, this is the first long-term prospective follow-up study reporting an independent protective role of higher folate status on several aspects of the natural history of high-risk HPV after controlling for known risk factors and other micronutrients. Improving folate status in subjects at risk of getting infected or already infected with high-risk HPV may have a beneficial impact in the prevention of cervical cancer.

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Section: Discussionmentioning

confidence: 99%

Folate Is Associated with the Natural History of High-Risk Human Papillomaviruses

Piyathilake

Henao²,

Macaluso³

et al. 2004

Cancer Research

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“…Besides, the use of these consensus primers may leave some important oncogenic HPV types undetected because of loss of the L1 sequence during viral integration. In clinical practice this can be critical, since integrated HPV 16/18 DNA has been revealed in various stages of dysplasia (Nagao et al, 2002;Gallo et al, 2003) and is shown to be common in patients with cervical carcinomas (Park et al, 1997;Kalantari et al, 1998;Walboomers et al, 1999). Therefore, the use of type-specific PCR, directed against one of the early-region genes (preferentially E6 and E7), is always recommended.…”

Section: Discussionmentioning

confidence: 99%

Human papillomavirus oncogenic expression in the dysplastic portio; an investigation of biopsies from 190 cervical cones

et al. 2004

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In this study, we investigated the presence of E6/E7 transcripts of seven common high-risk human papillomavirus (HPV) types in 190 cervical biopsies. The RNA-based real-time nucleic acid sequence-based amplification assay (NASBA) and type-specific PCR, both detecting HPV 16, 18, 31, 33, 45, 52, and 58, as well as consensus PCR, were performed on all 190 biopsies. High accordance between type-specific and consensus PCR confirms that the HPV types included in this study are the most common types present in cervical dysplasia. Furthermore, we see a clear increase in the incidence of HPV, both DNA and RNA, along with the histological severity of dysplasia. HPV RNA was detected in all but two PCR-positive cases, confirming that the virus exerts E6/E7 mRNA expression in cases of high-grade dysplasia. Out of 19 women given a normal or borderline diagnosis at conisation, only four were found HPV positive, which may suggest that unnecessary conisations can possibly be reduced by introducing HPV testing into the preoperative routine assessment. Human papillomavirus (HPV) infections are associated with the development of cervical neoplasia and are a necessary factor in the aetiology of cervical carcinoma (Walboomers et al, 1999;Bosch et al, 2002;zur Hausen, 2002). The most frequent HPV types found in high-grade cervical intraepithelial dysplasia (CIN II/III) and in cervical carcinomas are HPV 16, 18, 31, 33, and 45, which are often referred to as high-risk HPV or cancer-associated HPV types. However, the association between a positive HPV DNA test and the risk of subsequent development of dysplasia is partly unknown; the presence of HPV DNA does not necessarily indicate a risk for developing a more severe lesion. In fact, HPV is a very common virus among sexually active women and most infections are transient and asymptomatic (zur Hausen, 1991;Ho et al, 1998;Moscicki et al, 1998). Only very few infections cause morphologic changes in the epithelium and only a small percent of these will eventually develop cervical cancer (Bosch et al, 1995). On the other hand, several studies have shown that in cervical carcinogenesis, the expression of HPV E6 and E7 ORF is required for cell transformation and immortalisation (zur Hausen, 1988;Munger et al, 1989;Stoler et al, 1992). Consequently, persistent expression of these oncogenes may serve as an indicator of progression to CIN and invasive cancer (Sotlar et al, 1998), and detection of E6 and E7 mRNA transcripts may therefore be of higher prognostic value than the detection of HPV DNA. Moreover, new methods for preservation of RNA in cells have made gene expression analysis more feasible.In this study, we investigated the presence of high-risk HPV E6/E7 transcripts in cervical biopsies from patients undergoing conisation. The analysis was performed on a biopsy taken from the ectocervix/transformation zone at the time of conisation, and the pathological diagnosis of this particular biopsy was not always representative for the existing lesion that was later revealed in the cone. This s...

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“…In recent studies using real-time PCR (Nagao et al, 2002;Peitsaro et al, 2002), the integration of the genome of high-risk HPVs was detected not only in cervical carcinomas but also in CINs. Peitsaro et al (2002) showed that 14 of 15 HPV-16 -positive CIN1/2s contained HPV-16 DNA in either integrated or mixed form.…”

Section: Discussionmentioning

confidence: 99%

“…An earlier study using 2-dimensional gel electrophoresis showed that integration of high-risk HPV was characteristic of cervical carcinoma but not of CIN (Das et al, 1992). However, recent studies using real-time PCR suggested that integration of high-risk HPV into the host genome occurs in a fraction of cases of CIN (Nagao et al, 2002;Peitsaro et al, 2002). The factors that influence the HPV-mediated progression of dys-plastic lesions to invasive carcinoma have not been determined.…”

mentioning

confidence: 97%

Monoclonal Expansion with Integration of High-Risk Type Human Papillomaviruses Is an Initial Step for Cervical Carcinogenesis: Association of Clonal Status and Human Papillomavirus Infection with Clinical Outcome in Cervical Intraepithelial Neoplasia

Ueda

Enomoto

Miyatake

et al. 2003

Laboratory Investigation

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SUMMARY:To define the natural history of cervical intraepithelial neoplasia (CIN) as related to clonal status, we evaluated 20 cases of CIN1 and 18 cases of CIN2 that had been clinically followed for 7 to 48 months at Osaka University Hospital. These included 10 cases that progressed, 15 cases that persisted, and 13 cases that regressed. We analyzed the clonal status of each case by analysis of the pattern of X-chromosomal inactivation. Human papillomavirus (HPV) infection was detected by PCR-RFLP analysis. CINs that are monoclonal or infected by high-risk HPVs are more likely to progress or persist than cases that are polyclonal or negative for high-risk HPVs (p ϭ 0.009 for monoclonal vs polyclonal, p ϭ 0.024 for high-risk HPV positive vs negative p ϭ 0.024). Eighteen (90%) of 20 monoclonal, high-risk HPV-associated CINs progressed or persisted, whereas 9 (60%) of 15 polyclonal or high-risk HPV-negative CINs regressed. Therefore, the combination of clonality status and high-risk type HPV infection was significantly correlated with clinical outcome (p ϭ 0.003). The physical status of the HPV genome was evaluated in 17 cases of HPV-16 positive CINs by real-time PCR. Of those, the HPV viral genome was present in both episomal and integrated forms in 14 CINs (84%), and 12 of these cases (86%) were monoclonal in composition. By contrast, all three CINs in which the HPV genome was present in episomal form were polyclonal. In one CIN1 that was polyclonal, HPV-16 was originally present in episomal form but after 24 months, the patient developed a monoclonal CIN3 in which the HPV-16 genome was present in mixed form. These results may imply that HPV viral integration into the host genomic DNA is associated with progression from polyclonal to monoclonal status in CIN. These events may play a fundamental role in the progression from low-grade to higher grade dysplasia of the cervical mucosa. (Lab Invest 2003, 83:1517-1527.

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Rapid and Sensitive Detection of Physical Status of Human Papillomavirus Type 16 DNA by Quantitative Real-Time PCR

Cited by 84 publications

References 23 publications

Folate Is Associated with the Natural History of High-Risk Human Papillomaviruses

Folate Is Associated with the Natural History of High-Risk Human Papillomaviruses

Human papillomavirus oncogenic expression in the dysplastic portio; an investigation of biopsies from 190 cervical cones

Monoclonal Expansion with Integration of High-Risk Type Human Papillomaviruses Is an Initial Step for Cervical Carcinogenesis: Association of Clonal Status and Human Papillomavirus Infection with Clinical Outcome in Cervical Intraepithelial Neoplasia

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