Increased detection rate of human papillomavirus in cervical scrapes by the polymerase chain reaction as compared to modified FISH and southern‐blot analysi

Melchers, Willem J. G.; Brule, A. van den; Walboomers, Jan M. M.; Bruin, M. De; Burger, M. P. M.; Herbrink, Paul; Meijer, Chris J.L.M.; Lindeman, J.; Quint, Wim

doi:10.1002/jmv.1890270413

Cited by 123 publications

(55 citation statements)

References 24 publications

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“…Samples positive for HR HPV by the hc2 assay (Digene, Milan, Italy) were analyzed by a multiplex PCR assay using type-specific primer sequences as described by van den Brule et al (24,42) To assess sensitivity, plasmids containing genomic DNAs of HPV-16 and -18, kindly provided by E. M. de Villiers (Deutsches Krebsforschungszentrum, Heidelberg, Germany), and specific PCR products of HPV-31 and -33 cloned into the pCR2.1 vector (Invitrogen, San Diego, CA) were quantified by optical density measurement. Constructs were serially diluted and subsequently amplified using type-specific primers.…”

Section: Methodsmentioning

confidence: 99%

RNA (E6 and E7) Assays versus DNA (E6 and E7) Assays for Risk Evaluation for Women Infected with Human Papillomavirus

Cattani¹,

Siddu²,

D'Onghia³

et al. 2009

J Clin Microbiol

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In the majority of cases, high-risk human papillomavirus (HR HPV) infections regress spontaneously, with only a small percentage progressing to high-grade lesions. Current screening methods are based on DNA detection. An alternative would be to monitor expression of the E6 and E7 viral oncogenes continuously expressed by malignant phenotypes. In the work reported in this paper, we compared the two methods for a group of women with high-risk HPV infections. Cervical specimens from 400 women, previously found to be HPV DNA positive, were analyzed for HPV DNA by a liquid hybridization assay and typed by multiplex PCR (for types 16, 18, 31, and 33). Identification of HR HPV E6 and E7 RNA transcripts was performed using real-time reverse transcription-PCR and nucleic acid sequence-based amplification assays. Results were compared with concurrent cytological data. HR HPVs were found in 61.2% of patients. The most common genotype was HPV type 16 (HPV-16) (47.1%), followed by HPV-18, HPV-31, and HPV-33. Nine percent of cases involved other genotypes. Among 223 HPV DNA-positive samples, only 118 were positive in the RNA test. Among HPV DNA-positive patients with normal cytology, we detected E6 and E7 RNA transcripts in two cases (18.2%). The rate of detection increased gradually with the grade of the observed lesions, rising from 20% for patients with atypical squamous cells of undetermined significance to 48.1% for women with low-grade squamous intraepithelial lesions and 86.3% for those with high-grade squamous intraepithelial lesions. These results suggest that testing for HPV E6 and E7 transcripts could be a useful tool for screening and patient management, providing more accurate predictions of risk than those obtained by DNA testing.Cancer of the cervix is the second most frequent gynecological malignancy in the world. It is well established that the main cause is infection with human papillomavirus (HPV) (30,43,46,48). However, only certain specific types of virus lead to cancer. We can thus distinguish between high-risk and low-risk HPVs (HR HPV and LR HPV, respectively) (1,6,12,31,43). Epidemiological studies of HPV show that more than 70% of cervical cancers worldwide are caused by HPV type 16 (HPV-16) and HPV-18. The remaining cases of malignancy are associated with other types of HR HPV (mainly . Today, these viruses are among the most important known risk factors for human cancer (6,20,31,38).Most HR HPV infections regress spontaneously 6 to 12 months after their appearance, probably due to successful attack by the immune system (33). Only a small percentage of infections persist. Without surgical treatment, these infections can progress to high-grade lesions and to squamous cell carcinoma or adenocarcinoma of the cervix (37,47,48).The introduction of the Pap smear test by Papanicolaou made it possible to identify precursor lesions and to significantly reduce mortality. However, the test cannot reliably predict whether a mild dysplasia will regress or progress (5, 36). Recently, it was suggested that cervica...

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Section: Methodsmentioning

confidence: 99%

RNA (E6 and E7) Assays versus DNA (E6 and E7) Assays for Risk Evaluation for Women Infected with Human Papillomavirus

Cattani¹,

Siddu²,

D'Onghia³

et al. 2009

J Clin Microbiol

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“…The PCR was based on modified versions of the type-specific primer sequences described by van den Brule and colleagues (34,52 To assess sensitivity, plasmids containing genomic DNA of HPV-16 and HPV-18, kindly provided by E. M. de Villiers (Deutsches Krebsforschungszentrum, Heidelberg, Germany), and the specific PCR products of HPV-31, HPV-33, and HPV-45 cloned into the pCR2.1 vector (Invitrogen, San Diego, CA) were quantified by measurement of the optical density. The constructs were serially diluted and subsequently amplified with type-specific primers.…”

Section: Studymentioning

confidence: 99%

Clinical Performance of Human Papillomavirus E6 and E7 mRNA Testing for High-Grade Lesions of the Cervix

et al. 2009

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Infection with high-risk (HR) human papillomavirus (HPV) is the major cause of cervical cancer. However, relatively few infections progress to malignant disease. Progression to malignancy requires the overexpression of the E6 and E7 genes in the integrated HPV genome. It follows that the E6 and E7 transcripts could be useful markers of disease progression. The study presented here tests this possibility, using data from colposcopy and from cytological and histological tests to compare RNA assays for the E6 and E7 genes with DNA testing. A total of 180 women underwent colposcopy, cytology, and biopsy of suspected lesions (143 cases). Cervical brush specimens were analyzed for HPV DNA and for E6 and E7 mRNA. DNA from HR HPV was found in 57.8% of the specimens; E6 and E7 transcripts were found in 45%. The rates of detection of HPV DNA and of E6 and E7 transcripts were 33.3% and 25%, respectively, for specimens with normal findings; 51.4% and 31.9%, respectively, for specimens with cervical intraepithelial neoplasia grade 1 (CIN1); and 61.1% and 44.2% for specimens with CIN2, respectively. All specimens with CIN3 and 95.5% of specimens from patients with squamous cell carcinoma were positive by both assays. Thirty-seven patients with normal colposcopy findings did not undergo biopsy. HPV DNA and mRNA transcripts were found in 32.4% and 18.9% of these cases, respectively. Comparisons with cytological tests produced similar results. Overall, the mRNA tests showed a higher specificity than the DNA tests for high-grade lesions (72.7% and 56.2%, respectively) and a higher positive predictive value (59.3% and 49.0%, respectively). These findings suggest that mRNA assays could be more powerful than DNA testing for predicting the risk of progression and offer a strong potential as a tool for triage and patient follow-up.Carcinomas of the anogenital tract, particularly cancer of the cervix, represent the second most frequent type of neoplasm worldwide (43, 57). The major cause of these cancers is infection with high-risk (HR) human papillomavirus (HPV). DNA from HPV has been detected in more than 99% of cervical squamous cell carcinomas (SCCs) and a smaller proportion of adenocarcinomas (3,4,31,33). However, most HPV infections regress spontaneously or progress only after a long period of latency. As a result, the number of infections is far higher than the number of women who develop cancer.The most common types of HPV found in cancer patients are types 16, 18, 31, 33, and 45 (5, 10, 40, 53). Persistent infection with these types is regarded as a significant risk factor (40). The role of HPVs in the etiology of cervical cancer is tightly correlated with the overexpression of two oncogenes (E6 and E7) due to a specific opening in the E2 open reading frame in the integrated viral genome (23, 28). Studies of cervical cancer cell lines and cancer biopsy specimens have shown that the continuous expression of the genes is a necessary condition for the transformation and maintenance of neoplastic and dysplastic cells (46,56,57).Ce...

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“…segments of the L1 region, respectively. 13 SiHa cells containing one to two HPV 16 copies/cell, CaSki cells containing 600 HPV 16 copies/cell and HeLa53 cells containing 10-50 HPV 18 copies/cell were used as positive controls. Amplifications were performed in 50 µL volumes by using the same procedures for all primer sets.…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

Human papillomavirus associated with bladder carcinoma? Analysis by polymerase chain reaction

et al. 1999

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Background: The aim of the present study was to assess the possible etiologic role of human papillomaviruses (HPV) in bladder tumors. Methods: Forty-two fresh biopsy specimens from different grades and stages of bladder tumor cases and 10 normal bladder mucosa biopsies were studied. Specimens were analyzed by polymerase chain reaction (PCR) with HPV-specific general primer set for the detection of viral DNA. Polymerase chain reaction-positive samples were also tested with HPV 16-and 18-specific primers by the same method. Results: We found two samples (4.8%) containing HPV DNA among the TaG1 bladder tumors. All other specimens, including the control group, were found to be negative by PCR. Neither of the two HPV-positive patients had immune deficiency and/or genital warts. Human papillomavirus 16 was detected by type-specific primers in one sample, but the other HPV-positive sample could not be typed. Conclusions:The low prevalence of HPV in this and many previous studies does not support an etiologic role of HPV in bladder carcinogenesis. We detected the virus in two early stage tumors, but none was detected in the high-grade samples. However, to clarify the positivity of HPV in these occasional cases, future studies must be designed by using in situ PCR techniques, including samples from tumors and normal bladder mucosa from the same patient.

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Increased detection rate of human papillomavirus in cervical scrapes by the polymerase chain reaction as compared to modified FISH and southern‐blot analysi

Cited by 123 publications

References 24 publications

RNA (E6 and E7) Assays versus DNA (E6 and E7) Assays for Risk Evaluation for Women Infected with Human Papillomavirus

RNA (E6 and E7) Assays versus DNA (E6 and E7) Assays for Risk Evaluation for Women Infected with Human Papillomavirus

Clinical Performance of Human Papillomavirus E6 and E7 mRNA Testing for High-Grade Lesions of the Cervix

Human papillomavirus associated with bladder carcinoma? Analysis by polymerase chain reaction

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