scholarly journals Persistence of Human Papillomavirus DNA in Cervical Lesions After Treatment With Diathermic Large Loop Excision

1998 ◽  
Vol 6 (5) ◽  
pp. 214-219 ◽  
Author(s):  
A. L. Distéfano ◽  
M. A. Picconi ◽  
L. V. Alonio ◽  
D. Dalbert ◽  
J. Mural ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Blot Hybridization ◽  
Simultaneous Detection ◽  
Cervical Lesions ◽  
Dot Blot ◽  
Hpv Dna ◽  
Viral Type ◽  
Large Loop ◽  
Single Stranded Conformational Polymorphism

Objective:The aim of this study was to identify human papillomavirus (HPV) in cervical intraepithelial neoplasia (CIN) lesions and to evaluate the persistence of viral DNA after diathermic large loop excision (DLLE) treatment.Study Design:Biopsies from 36 patients with low- and high-grade CIN lesions were studied before and after DLLE treatment looking for HPV sequences. DNA was extracted to perform a radioactive polymerase chain reaction (PCR) using GP 5,6 generic primers. PCR products were analyzed by the single-stranded conformational polymorphism (SSCP) which is a simultaneous detection and typing method. Dot-blot hybridization with generic and type-specific biotinylated oligonucleotide probes was applied in some cases.Results:HPV DNA was found in all pretreatment samples, and the viral type was identified in 80% of them, HPV 16 being the most prevalent. The viral type coincided with that detected in the first biopsy in all except one case. Seventy five percent of the patients (27 cases) were negative for CIN at follow up, but 50% of them remained HPV DNA positive.Conclusion:DLLE treatment was effective in removing the CIN lesion but not the HPV. This fact points out the need to asses the presence of HPV in DNA during the follow-up, since viral persistence has been considered a high risk factor for recurrence and/or malignant transformation.

scholarly journals Genotyping of 27 Human Papillomavirus Types by Using L1 Consensus PCR Products by a Single-Hybridization, Reverse Line Blot Detection Method

1998 ◽  
Vol 36 (10) ◽  
pp. 3020-3027 ◽  
Author(s):  
P. E. Gravitt ◽  
C. L. Peyton ◽  
R. J. Apple ◽  
C. M. Wheeler
Keyword(s):  
Human Papillomavirus ◽  
Detection System ◽  
Sampling Error ◽  
Blot Hybridization ◽  
High Sensitivity ◽  
Oligonucleotide Probes ◽  
Dot Blot ◽  
Consensus Primer ◽  
Hpv Dna ◽  
Silver Spring

Amplification of human papillomavirus (HPV) DNA by L1 consensus primer systems (e.g., MY09/11 or GP5+/6+) can detect as few as 10 to 100 molecules of HPV targets from a genital sample. However, genotype determination by dot blot hybridization is laborious and requires at least 27 separate hybridizations for substantive HPV-type discrimination. A reverse blot method was developed which employs a biotin-labeled PCR product hybridized to an array of immobilized oligonucleotide probes. By the reverse blot strip analysis, genotype discrimination of multiple HPV types can be accomplished in a single hybridization and wash cycle. Twenty-seven HPV probe mixes, two control probe concentrations, and a single reference line were immobilized to 75- by 6-mm nylon strips. Each individual probe line contained a mixture of two bovine serum albumin-conjugated oligonucleotide probes specific to a unique HPV genotype. The genotype spectrum discriminated on this strip includes the high-risk, or cancer-associated, HPV genotypes 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 55, 56, 58, 59, 68 (ME180), MM4 (W13B), MM7 (P291), and MM9 (P238A) and the low-risk, or non-cancer-associated, genotypes 6, 11, 40, 42, 53, 54, 57, 66, and MM8 (P155). In addition, two concentrations of β-globin probes allowed for assessment of individual specimen adequacy following amplification. We have evaluated the performance of the strip method relative to that of a previously reported dot blot format (H. M. Bauer et al., p. 132–152, in C. S. Herrington and J. O. D. McGee (ed.), Diagnostic Molecular Pathology: a Practical Approach, (1992), by testing 328 cervical swab samples collected in Digene specimen transport medium (Digene Diagnostics, Silver Spring, Md.). We show excellent agreement between the two detection formats, with 92% concordance for HPV positivity (kappa = 0.78, P < 0.001). Nearly all of the discrepant HPV-positive samples resulted from weak signals and can be attributed to sampling error from specimens with low concentrations (<1 copy/μl) of HPV DNA. The primary advantage of the strip-based detection system is the ability to rapidly genotype HPVs present in genital samples with high sensitivity and specificity, minimizing the likelihood of misclassification.

Human papillomavirus in cervix carcinoma and condylomata acuminate-identification of HPV-DNA by improved dot-blot-hybridization

1992 ◽  
Vol 17 (6) ◽  
pp. 392-396 ◽  
Author(s):  
R.M. KNOBLER ◽  
S. SCHNEIDER ◽  
B. RADLWIMMER ◽  
W. BODEMER ◽  
W. GEBHART ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Blot Hybridization ◽  
Cervix Carcinoma ◽  
Dot Blot ◽  
Hpv Dna ◽  
Dot Blot Hybridization

Human Papillomavirus (HPV) Cervical Lesions: Results from 300 Italian Women Studied with Dna Hybridization Techniques and Morphology

1988 ◽  
Vol 74 (6) ◽  
pp. 745-749 ◽  
Author(s):  
Fiorella Nuzzo ◽  
Vittorio Tison ◽  
Antonella Castagnoli ◽  
Maurizio Tiboni ◽  
Ethel-Michele De Villiers ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Human Papillomavirus ◽  
Southern Blot ◽  
Dna Hybridization ◽  
Blot Hybridization ◽  
Hpv Infection ◽  
Cervical Lesions ◽  
Hpv Dna ◽  
Cervical Infection

Human papillomavirus cervical infection was investigated in a series of 300 unselected women by comparing morphological diagnoses (cytology and histology) with results of DNA hybridization techniques (filter in situ hybridization of DNA from exfoliated cervical cells and Southern blot analysis of HPV-DNA in cervical biopsy specimens). The prevalence of HPV cervical infection diagnosed by PAP smears was 11.6 %. Despite disadvantages, filter in situ hybridization was confirmed to be particularly useful for screening purposes to detect HPV in cervical scrapings. In 3 cases it was the only applicable method for diagnosing « high-risk » HPV infection. Southern blot hybridization of tissue DNA with HPV 16-DNA revealed the presence of this virus in 8 cases, and HPV 31-DNA and HPV 42-DNA in 1 case each.

scholarly journals Molecular variants of human papillomavirus types 16 and 18 preferentially associated with cervical neoplasia

2000 ◽  
Vol 81 (12) ◽  
pp. 2959-2968 ◽  
Author(s):  
Luisa L. Villa ◽  
Laura Sichero ◽  
Paula Rahal ◽  
Otavia Caballero ◽  
Alex Ferenczy ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Cancer Screening ◽  
Screening Program ◽  
Blot Hybridization ◽  
Cervical Neoplasia ◽  
Long Control Region ◽  
Cervical Lesions ◽  
Dot Blot ◽  
Hpv 16 ◽  
Molecular Variants

In order to determine geographically related intratypic variation in human papillomavirus (HPV) type 16 and 18 isolates that could be associated with lesion development, data were analysed from an ongoing cohort study of the natural course of infection of HPVs and cervical neoplasia. Testing for HPVs was carried out by PCR and molecular variants of these HPVs were characterized by sequence analysis of the long control region and by dot blot hybridization of the E6 and L1 genes. Tests for HPV were done in multiple first-year specimens from 1690 women enrolled in a cancer screening program from 1993 to 1997. Subjects were followed-up by cytology and cervicography for detection of cervical lesions. Seven variants of HPV-16 and four of HPV-18 were detected in one or more specimens from 65 subjects. The same variant was found in specimens taken on different visits from each case of persistent infection. Overall, non-European variants tended to persist more frequently [odds ratio (OR)=4·5; 95% confidence interval (CI), 1·6–12·4] than European (E) variants (OR=2·5; 95% CI, 1·3–4·9), relative to the risk of persistence for non-oncogenic HPVs. In addition, non-E variants were more strongly associated with risk of both prevalent (age- and race-adjusted OR=172·2; 95% CI, 47·1–630·1) and incident [relative risk (RR)=22·5; 95% CI, 6·0–83·9] high-grade lesions than E variants (prevalent lesions OR=46·3; 95% CI, 15·5–138·0 and incident lesons RR=6·1; 95% CI, 1·3–27·4), relative to the risk for HPV-negative women. Although consistent, the latter differences were not statistically significant. If confirmed in other populations, measurement of intratypic variation of HPV-16 and -18 has the potential to serve as an ancillary tool in cervical cancer screening.

Molecular detection methods of human papillomavirus (HPV)

2009 ◽  
Vol 24 (4) ◽  
pp. 215-222 ◽  
Author(s):  
Apostolos Zaravinos ◽  
Ioannis N. Mammas ◽  
George Sourvinos ◽  
Demetrios A. Spandidos
Keyword(s):  
Human Papillomavirus ◽  
Molecular Detection ◽  
Blot Hybridization ◽  
High Sensitivity ◽  
Detection Methods ◽  
Dot Blot ◽  
Hpv Dna ◽  
Moderate Sensitivity ◽  
Hybridization Assays

Human papillomavirus (HPV) testing can identify women at risk of cervical cancer. Currently, molecular detection methods are the gold standard for identification of HPV. The three categories of molecular assays that are available are based on the detection of HPV DNA and include (1) non-amplified hybridization assays, such as Southern transfer hybridization (STH), dot blot hybridization (DB) and in situ hybridization (ISH); (2) signal amplified hybridization assays, such as hybrid capture assays (HC2); (3) target amplification assays, such as polymerase chain reaction (PCR) and in situ PCR. STH requires large amounts of DNA, is laborious and not reproducible, while ISH has only moderate sensitivity for HPV. The sensitivity of the HC2 assay is similar to that of PCR-based assays, with high sensitivity being achieved by signal rather than target amplification. PCR-based detection is both highly sensitive and specific. Since PCR can be performed on very small amounts of DNA, it is ideal for use on specimens with low DNA content. In the future, with the advance of technology, viral DNA extraction and amplification systems will become more rapid, more sensitive, and more automated.

Genital Human Papillomavirus Infections: Correlation of Cytological, Colposcopic and Histological Features with Viral Types in Women and Their Male Partners

1993 ◽  
Vol 4 (1) ◽  
pp. 13-20 ◽  
Author(s):  
J Monsonego ◽  
L Zerat ◽  
F Catalan ◽  
Y Coscas
Keyword(s):  
Human Papillomavirus ◽  
Blot Hybridization ◽  
Diagnostic Methods ◽  
Low Grade ◽  
Male Partners ◽  
Papillomavirus Infections ◽  
Cervical Lesions ◽  
Hpv 16 ◽  
Histological Features ◽  
Hpv Dna

To determine the incidence of anogenital papillomavirus infections and to assess the value of available diagnostic methods, we compared the cytological, colposcopic and histological features of anogenital papillomavirus-related lesions with their associated human papillomavirus types (HPV) in 300 women and in their male partners. HPV-type deoxyribonucleic acid was detected by blot hybridization in 398 out of 624 subclinical and clinically defined anogenital lesions. Whatever the site of the lesion, condylomas and low-grade intraepithelial neoplasia (IEN) were found in 84% of lesions associated with HPV 6–11, compared with 32% of lesions containing HPV 16–18 ( P<0.001). Among the HPV 16–18 associated lesions, high-grade cervical, vaginal, vulvar and anal intraepithelial neoplasias represented 45% ( P<0.001) of the lesions. In 65% of 23 cases of squamous anogenital cancer, HPV 16–18 and mixed types were present ( P<0.001). In 54% (161/300) of cases, the lesions were multicentric (161/300). On cytological examination, 27% of the samples gave false negative results. In cervical lesions, there was a good correlation between virological and colposcopic findings, but this was not true for extracervical mucous epithelia in the vagina or on the vulva. With peniscopy in the male partners 220 out of 410 had penile condylomatous lesions and more than half of the 350 male specimens examined by molecular hybridization contained HPV DNA. A correlation was found between the virus types in penile lesions or in cells of the distal urethra and in the cervical lesions of the sexual partner. We concluded that in the majority of cases of cervical lesions there is a correlation between the type of HPV DNA identified by blot hybridization and the cytohistocolposcopic findings. In practice, viral typing may be indicated in cases of colpohistologic discordance and in condyloma low-grade CIN that cannot be distinguished from presumably innocuous human papillomavirus infection.

scholarly journals A Matched Prospective Study of Human Immunodeficiency Virus Serostatus, Human Papillomavirus DNA, and Cervical Lesions Detected by Cytology and Colposcopy

1999 ◽  
Vol 7 (3) ◽  
pp. 158-164 ◽  
Author(s):  
L. O. Eckert ◽  
D. H. Watts ◽  
L. A. Koutsky ◽  
S. E. Hawes ◽  
C. E. Stevens ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Human Papillomavirus ◽  
Prospective Study ◽  
Cervical Lesions ◽  
Abnormal Cytology ◽  
Hpv Dna ◽  
Immunodeficiency Virus ◽  
Dna Types ◽  
Hiv Seropositive

Objective:To compare the prevalence and type of human papillomavirus (HPV) infections in the genital tract of human-immunodeficiency-virus- (HIV) seropositive and -seronegative women matched for cytology and to examine prospectively the relationship of HPV DNA, colposcopic findings and cervical squamous intraepithelial lesions (SIL) in these matched seropositive and seronegative cohorts.Methods:A matched prospective study of HIV-seropositive and -seronegative women undergoing cytologic screening, colposcopy, and testing for HPV DNA and other infections at each visit.Results:Twenty-three HIV-seropositive women were matched with 23 seronegative women by cervical cytology reading, lifetime number of sexual partners, age, and follow-up length. Fourteen pairs of these women had follow-up visits every 4 months, for 56 and 53 total visits in seropositive and seronegative women, respectively. After matching, the groups had a similar overall prevalence ofHPV DNA and ofHPV oncogenic (high risk) types at baseline. On follow up, HIV-seropositive women were more likely than seronegative women to develop SIL (38% vs. 10%), less likely to have negative cytology (34% vs. 60%, overallP= 0.03), more visits with HPV DNA detected (68% vs. 40%P= 0.04), and more visits with multiple HPV DNA types detected (18% vs. 0%,P= 0.02). Colposcopic lesions in the seropositive women were more likely to have sharp borders or mosaicism or to be thick white (P= 0.009).Conclusions:After matching for baseline Papanicolaou smear readings, these data suggest that over time seropositive women have more visits that yield abnormal cytology, more persistent HPV DNA detection, and more colposcopic abnormalities than seronegative women. Infect. Dis. Obstet. Gynecol. 7:158–164, 1999.

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