Molecular diagnosis of genital human papillomavirus infection: Comparison of two methods used to collect exfoliated cervical cells

Vermund, Sten H.; Schiffman, Mark; Goldberg, Gary L.; Ritter, Diane B.; Weltman, André; Burk, Robert D.

doi:10.1016/0002-9378(89)90430-4

Cited by 57 publications

(20 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus, the sensitivity of PCR and LCR could have been higher if only a urine sample (as would happen in a screening program) had been taken. The order in which specimens were taken in a study comparing two methods for diagnosing genital human papillomavirus infection was evaluated by Vermund et al and showed slight influence on the diagnostic accuracy (20). On the other hand, to the extent that one is interested in evaluating the performance of a procedure that can be broadly applied to asymptomatic men, compared to a procedure that can only be applied within the clinic setting, the comparisons made in this analysis are appropriate.…”

Section: Discussionmentioning

confidence: 99%

Relative Accuracy of Nucleic Acid Amplification Tests and Culture in Detecting Chlamydia in Asymptomatic Men

Cheng¹,

Macaluso²,

Vermund³

et al. 2001

J Clin Microbiol

Self Cite

View full text Add to dashboard Cite

Published estimates of the sensitivity and specificity of PCR and ligase chain reaction (LCR) for detecting Chlamydia trachomatis are potentially biased because of study design limitations (confirmation of test results was limited to subjects who were PCR or LCR positive but culture negative). Relative measures of test accuracy are less prone to bias in incomplete study designs. We estimated the relative sensitivity (RSN) and relative false-positive rate (RFP) for PCR and LCR versus cell culture among 1,138 asymptomatic men and evaluated the potential bias of RSN and RFP estimates. PCR and LCR testing in urine were compared to culture of urethral specimens. Discordant results (PCR or LCR positive, but culture negative) were confirmed by using a sequence including the other DNA amplification test, direct fluorescent antibody testing, and a DNA amplification test to detect chlamydial major outer membrane protein. The RSN estimates for PCR and LCR were 1.45 (95% confidence interval [CI] ‫؍‬ 1.3 to 1.7) and 1.49 (95% CI ‫؍‬ 1.3 to 1.7), respectively, indicating that both methods are more sensitive than culture. Very few false-positive results were found, indicating that the specificity levels of PCR, LCR, and culture are high. The potential bias in RSN and RFP estimates were <5 and <20%, respectively. The estimation of bias is based on the most likely and probably conservative parameter settings. If the sensitivity of culture is between 60 and 65%, then the true sensitivity of PCR and LCR is between 90 and 97%. Our findings indicate that PCR and LCR are significantly more sensitive than culture, while the three tests have similar specificities.

show abstract

Section: Discussionmentioning

confidence: 99%

Relative Accuracy of Nucleic Acid Amplification Tests and Culture in Detecting Chlamydia in Asymptomatic Men

Cheng¹,

Macaluso²,

Vermund³

et al. 2001

J Clin Microbiol

Self Cite

View full text Add to dashboard Cite

show abstract

“…The vaginal tampon was placed in a sterile jar containing 50 ml of 10 mM Tris-HCl (pH 7.5), 50 mM EDTA, and 150 mM NaCl (9). During the pelvic examination, a cervicovaginal lavage was obtained with 10 ml of phosphate-buffered saline (pH 7.4) sprayed on the ectocervix with a syringe and aspirated from the posterior vaginal fornix (4,18). Specimens were refrigerated within 1 h. The delay between sampling and processing never exceeded 7 days.…”

Section: Methodsmentioning

confidence: 99%

“…Amplifications were performed in a TC9600 thermocycler (Perkin-Elmer Cetus, Montréal, Canada) for 40 cycles with the following cycling parameters: 95°C for 1 min, 55°C for 1 min, and 72°C for 1 min. Amplified products were spotted onto nylon membranes and were hybridized under stringent conditions as described in previous publications with 32 P-labeled oligonucleotide probes for types 6,11,16,18,31,33,35,39,45, 51, 52, 53, 56, and 58 (1,2,16). PCR products spotted onto nylon membranes were also hybridized with an HPV generic probe mixture under low-stringency conditions as previously described (1, 2, 14, 16).…”

Section: Methodsmentioning

confidence: 99%

“…However, screening samples for the presence of additional HPV types still requires gel electrophoretic analysis or generic probe blotting. We describe here a simple method for a broad-spectrum HPV screening assay; the method uses a generic probe mix composed of digoxigenin (DIG)-labeled fragments from four HPV types (11,16,18, and 51) on microwell plates (MWP) and a DIG-MWP detection kit from Roche Molecular Biochemicals. The assay utilizes the same biotinylated amplification products used in the MY09/ MY11/HMB01 reverse line blot genotyping techniques, eliminating the need for additional PCR.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Nonisotopic Detection of Human Papillomavirus DNA in Clinical Specimens Using a Consensus PCR and a Generic Probe Mix in an Enzyme-Linked Immunosorbent Assay Format

et al. 2001

View full text Add to dashboard Cite

We assessed the value of a new digoxigenin (DIG)-labeled generic probe mix in a PCR-enzyme-linked immunosorbent assay format to screen for the presence of human papillomavirus (HPV) DNA amplified from clinical specimens. After screening with this new generic assay is performed, HPV DNA-positive samples can be directly genotyped using a reverse blotting method with product from the same PCR amplification. DNA from 287 genital specimens was amplified via PCR using biotin-labeled consensus primers directed to the L1 gene. HPV amplicons were captured on a streptavidin-coated microwell plate (MWP) and detected with a DIG-labeled HPV generic probe mix consisting of nested L1 fragments from types 11, 16, 18, and 51. Coamplification and detection of human DNA with biotinylated ␤-globin primers served as a control for both sample adequacy and PCR amplification. All specimens were genotyped using a reverse line blot assay (13). Results for the generic assay using MWPs and a DIG-labeled HPV generic probe mix (DIG-MWP generic probe assay) were compared with results from a previous analysis using dot blots with a radiolabeled nested generic probe mix and type-specific probes for genotyping. The DIG-MWP generic probe assay resulted in high intralaboratory concordance in genotyping results (88% versus 73% agreement using traditional methods). There were 207 HPV-positive results using the DIG-MWP method and 196 positives using the radiolabeled generic probe technique, suggesting slightly improved sensitivity. Only one sample failed to test positive with the DIG-MWP generic probe assay in spite of a positive genotyping result. Concordance between the two laboratories was nearly 87%. Approximately 6% of samples that were positive or borderline when tested with the DIG-MWP generic probe assay were not detected with the HPV type-specific panel, perhaps representing very rare or novel HPV types. This new method is easier to perform than traditional generic probe techniques and uses more objective interpretation criteria, making it useful in studies of HPV natural history.Some types of human papillomavirus (HPV) are widely accepted as causative agents for cervical cancer (3,19). There are more than 40 HPV viral types that are commonly found in the genital tract, and approximately one-third of these are associated with cervical cancer and anal neoplasia. The anogenital HPV types are generally categorized as being either "high risk" or "low risk." High-risk types are associated with high-grade precancerous lesions and invasive cancer, while low-risk types are found in asymptomatic or benign conditions such as genital warts. However, the distribution and prevalence of types vary somewhat by geographic region and other demographic factors. Because the significance of the variation in type distribution is still being elucidated, studies of HPV epidemiology need to employ a methodology that can detect the entire spectrum of viral types. One of the most common means to detect and characterize new HPVs has been by PCR using consensus primers...

show abstract

“…More than 60% of women with abnormal Pap smears in our cachement area were reported previously to be HPV-positive [25,26]. The definitive role of HPV and the interaction of antioxidant vitamins and active oxy gen species in the complex multifactorial etiology of cer vix cancer needs to be further evaluated.…”

Section: Discussionmentioning

confidence: 99%

Measurements of Ascorbic Acid and Glutathione in Exfoliated Cervicovaginal Epithelial Cells of Smokers and Women with Cervical Dysplasias

Basu¹,

Mikhail²,

Goldberg³

et al. 1990

Gynecol Obstet Invest

Self Cite

View full text Add to dashboard Cite

This report emphasizes the ability to quantify ascorbic acid (AA) and reduced glutathione (GSH) levels in exfoliated cervicovaginal epithelial cells obtained by a lavage technique. Sixty-two women with abnormal Papanicolaou smears underwent colposcopic examinations. Colposcopic lesions were biopsied and histopathologically graded. Marked variations in the number of cells and in the levels of AA and GSH were observed. In cigarette smokers, the number of exfoliated cells retrieved was significantly higher (p < 0.05, by Student’s t test). The simultaneous investigation of biochemical and virologic parameters in exfoliated cervicovaginal epithelial cells, in conjunction with the known cytopathologic and epidemiologic risk variables, provides a novel approach to elucidate factor(s) that may inhibit or promote cervical carcinogenesis in designed prospective studies.

show abstract

Molecular diagnosis of genital human papillomavirus infection: Comparison of two methods used to collect exfoliated cervical cells

Cited by 57 publications

References 8 publications

Relative Accuracy of Nucleic Acid Amplification Tests and Culture in Detecting Chlamydia in Asymptomatic Men

Relative Accuracy of Nucleic Acid Amplification Tests and Culture in Detecting Chlamydia in Asymptomatic Men

Nonisotopic Detection of Human Papillomavirus DNA in Clinical Specimens Using a Consensus PCR and a Generic Probe Mix in an Enzyme-Linked Immunosorbent Assay Format

Measurements of Ascorbic Acid and Glutathione in Exfoliated Cervicovaginal Epithelial Cells of Smokers and Women with Cervical Dysplasias

Contact Info

Product

Resources

About