Factors associated with HPV persistence after treatment for high-grade cervical intra-epithelial neoplasia with large loop excision of the transformation zone (LLETZ).

BACKGROUND AND OBJECTIVE: Human Papillomavirus (HPV) persistence after high-grade cervical intra-epithelial neoplasia (CIN) removal may be associated with residual lesions or risk of disease recurrence. Knowledge regarding the factors associated with HPV persistence following CIN treatment is still limited. The main purpose of this longitudinal study was to assess the association between characteristics of the patients and their cervical lesions with high-risk HPV-type persistence, detected by commercially available Hybrid Capture II (HC II), after CIN 2 and 3 treatment with large loop excision of the transformation zone (LLETZ). STUDY DESIGN: For this cohort study, a total of 94 women submitted to LLETZ between March 2001 and September 2002 were included. Only women with at least one follow-up visit at 6 or 12 months and confirmed CIN 2 or 3 in the cone specimen were considered. In each visit women answered to a questionnaire and undertook Pap smear and HC II specimens collection. McNemar's, chi-square and Fisher tests were used for univariate analysis. Generalized Estimating Equations (GEE) were used for multivariate analysis. All calculations were performed within 95% confidence intervals (95% CI). RESULTS: Histological evaluation showed 12 (13%) women with CIN, 2 and 82 (87%) with CIN 3 and conization margins were compromised in 27 (29%) cases. Eighty-seven (92%) women showed positive HC II tests prior to LLETZ. Of women initially HPV negative, none had a positive HC II during follow-up. The proportion of positive HPV tests was reduced from 92% to 20%(P < 0.01) at the first visit and to 22% (P < 0.01) at the second visit after LLETZ. Multivariate analysis showed that smoking and age above 35 years (irrespective of margin status) were strongly associated with positive HPV during follow-up. CONCLUSION: HPV persistence following LLETZ was associated with smoking and with the interaction between age and conization margins.     

Frequency of cervical intraepithelial neoplasia following large loop excision of the transformation zone.

AIM: To determine the frequency of cervical intraepithelial neoplasia (CIN) following large loop excision of the transformation zone of the cervix (LLETZ) according to grade and completeness of excision of CIN. METHODS: A retrospective study of 669 patients who had LLETZ biopsies showing CIN 1, 2, or 3. The patients were subdivided according to the grade and completeness of excision of CIN. The follow up period was 1.5 to 3.5 years. Risk of persistent/recurrent CIN was assessed by the frequency of histological diagnosis of CIN during the follow up period. RESULTS: Frequency of persistent/recurrent CIN increased with the grade of CIN reported: 6.7% of patients with CIN 1, 13.4% with CIN 2, and 21.7% with CIN 3 developed persistence or recurrence. The frequency of CIN persistence/recurrence was significantly lower where LLETZ showed complete excision of CIN (8.4%) than where it was incomplete (31.3%) (p < 0.0001) or equivocal (27.8%) (p < 0.0001). CONCLUSIONS: Patients with incomplete or equivocal excision of all grades of CIN merit careful, preferably colposcopic, follow up. Patients with completely excised high grade CIN require careful cervical cytologic surveillance.     

Prevalence of the most common high-risk HPV genotypes among women in three new independent states of the former Soviet Union

Type distribution of HPV has been studied in different geographic regions, but the data are scanty from the new independent states of the former Soviet Union. Here the HPV prevalence and distribution of the most frequent high-risk HPV types among 3,187 women at different risk for HPV and cervical intraepithelial neoplasia in Russia, Belarus, and Latvia is reported. HPV detection, type distribution and viral load analysis in DNA samples from cervical scrapes were done with real-time PCR-based assay detecting HPV types 16, 18, 31, 33, 35, 39, 45, 52, and 58. The overall HPV prevalence was 31.2%, HPV16 was the most prevalent type followed by HPV31 and HPV33 group. The overall HPV prevalences in Russia, Belarus and Latvia were 33.4%, 27.5%, and 26.2%. The type distributions were similar in these countries, except for Latvia where HPV39 was the third prevalent genotype. HPV prevalence was highest (40.8%) among women from sexually transmitted disease clinic, followed by 30.9% among gynecological outpatients and 27.2% in screening patients. HPV detection increased with cytological abnormality (P = 0.0001) and lesion grade in the biopsy (P = 0.0001), from 27% to 72% in normal samples to cancer, and from 64% to 77% in cervical intraepithelial neoplasia 1 to cancer. The normalized viral loads varied greatly between and among different HPV-types. The mean log HPV33 group copies/cell increased from negative for intraepithelial lesions to cancer (P = 0.049). Distribution of the most common high-risk HPV-types seems to be similar in these countries as reported in other major geographical regions.     

Performance of the Abbott RealTime high‐risk HPV test in women with abnormal cervical cytology smears

HPV DNA testing is known to be much more sensitive than cytology, but less specific. A range of HPV and related tests in 858 women referred for colposcopy because of an abnormal smear were evaluated to compare the performances of these tests. This article compared the Abbott test to other tests which had been previously evaluated. This test was a real true test for 14 high‐risk HPV types. The Abbott test was found to be highly sensitive for cervical intraepithelial neoplasia grade 3 or worse (CIN3+) (98.9%) with a specificity of 31.5%. These numbers were comparable with the Qiagen HC2 test, the Roche Linear Array and Amplicor tests, and the Gen‐Probe APTIMA test. Differences between these tests appeared to be related mostly to the choice of cutoff level. An added feature of the Abbott test was the provision of type specific results for HPV 16 and 18. J. Med. Virol. 82: 1186–1191, 2010. © 2010 Wiley‐Liss, Inc.     

Detection of rare and possibly carcinogenic human papillomavirus genotypes as single infections in invasive cervical cancer

The contribution of carcinogenic human papillomavirus (HPV) types to the burden of cervical cancer has been well established. However, the role and contribution of phylogenetically related HPV genotypes and rare variants remains uncertain. In a recent global study of 8977 HPV‐positive invasive cervical carcinomas (ICCs), the genotype remained unidentified in 3.7% by the HPV SPF₁₀ PCR–DEIA–LiPA₂₅ (version 1) algorithm. The 331 ICC specimens with unknown genotype were analysed by a novel sequence methodology, using multiple selected short regions in L1. This demonstrated HPV genotypes that have infrequently or never been detected in ICC, ie HPV26, 30, 61, 67, 68, 69, 73 and 82, and rare variants of HPV16, 18, 26, 30, 34, 39, 56, 67, 68, 69, 82 and 91. These are not identified individually by LiPA₂₅ and only to some extent by other HPV genotyping assays. Most identified genotypes have a close phylogenetic relationship with established carcinogenic HPVs and have been classified as possibly carcinogenic by IARC. Except for HPV85, all genotypes in α‐species 5, 6, 7, 9 and 11 were encountered as single infections in ICCs. These species of established and possibly carcinogenic HPV types form an evolutionary clade. We have shown that the possibly carcinogenic types were detected only in squamous cell carcinomas, which were often keratinizing and diagnosed at a relatively higher mean age (55.3 years) than those associated with established carcinogenic types (50.9 years). The individual frequency of the possibly carcinogenic types in ICCs is low, but together they are associated with 2.25% of the 8338 included ICCs with a single HPV type. This fraction is greater than seven of the established carcinogenic types individually. This study provides evidence that possibly carcinogenic HPV types occur as single infections in invasive cervical cancer, strengthening the circumstantial evidence of a carcinogenic role. Copyright © 2012 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Prevalence of HPV genotypes determined by PCR and DNA sequencing in cervical specimens from French women with or without abnormalities

BACKGROUND: Epidemiological data on human papillomavirus (HPV) are needed to estimate potential changes in type distribution induced by recent HPV vaccination strategies. OBJECTIVES AND STUDY DESIGN: The epidemiological distribution of HPV in 669 cervical specimens from French women with and without cytological abnormalities was evaluated using type-specific PCR or sequencing. The results were compared with those obtained using the Digene high-risk Hybrid Capture 2 (HR-HC2) assay. RESULTS: The overall prevalence of HPV was high (45.3%) in our study population. 285 of the 291 HPV-positive samples were typed. The distribution frequency concerned 34 different genotypes, with HPV16 being the most prevalent (32.6%). Other genotypes present were HPV31 (7.4%), HPV18, HPV 52 (both 6.0%), HPV6 (5.3%) and HPV66 (4.2%). The respective frequencies of all other genotypes were below 4%. The agreement with HR-HC2 was 78.8%. The distribution frequency data were also analyzed relatively to cytological and histological results. Our method enables the diagnosis of HPV infections with the additional advantage of genotyping. CONCLUSION: HPV infections in the area of France studied here involve numerous HPV types, but the high cumulative prevalences of types 16, 18, 6 and 11 (44.6% in total) would suggest a major impact of vaccination on these genotypes.     

宫颈癌及其前期病变HPV感染与基因型分布

目的了解宫颈癌及宫颈上皮内瘤变CIN患者的HPV的感染和其基因分型及主要感染型别情况。方法应用型特异PCR检测宫颈癌及其前病变的患者的HPV感染及其主要基因分型情况的分析。结果在本研究宫颈癌及宫颈上皮内瘤变患者中,宫颈癌的HPV感染率为91．0％,CINⅠ／Ⅱ／Ⅲ的HPV感染率为73．3％,主要高危型HPV基因型别依次为HPV16、HPV18、HPV58、HPV33。结论在宫颈上皮内瘤变患者中感染主要高危型HPV基因型别依次为HPV16、HPV18、HPV58、HPV33、HPV16在宫颈癌和CIN中的构成比随着宫颈病变的增加而明显增加。     

ASC-US and high-risk HPV testing: performance in daily clinical practice

Data are beginning to accrue on high-risk HPV DNA testing in patients with ASC-US on cervical cytology. We report on our experience at the University of Wisconsin Hospital and Clinics. From February 2002 through December 31, 2005 (3 yr, 11 mo), the cytopathology laboratory processed 49,599 Pap Tests, of which 1,792 (3.6%) were diagnosed as ASC-US. Six hundred and seventy two (37.5%) of these cases were processed for high-risk HPV genotypes using the Digene Hybrid Capture II method. Of these cases, 266 (39.6%) were positive for high-risk HPV genotypes, 11 (1.6%) were equivocal, and 395 (58.8%) were negative. Biopsy follow-up was available for 127 (47.7%) of the 266 cases, of which 66 (52%) were negative, 46 (36.2%) showed CIN I, 9 (7.1%) were CIN II, and 6 (4.7%) were CIN III. Of the remaining 139 (52.3%) cases, 86 (62%) had follow-up Pap Tests, of which 57 (66.3%) were negative, 15 (17.4%) were ASC-US, 12 (15%) were low-grade squamous intraepithelial lesions, and 2 (2.3%) were high-grade squamous intraepithelial lesions; 53 (38.1%) were lost to follow-up. In combination, 90 (42.25%) of the 213 cases with follow-up showed atypia or above after a diagnosis of ASC-US; of which 58 (64%) were low-grade lesions and 17 (19%) were high-grade lesions. Our laboratory's reported high-risk HPV positivity is comparable to recent reports in the literature on its use in daily clinical practice. In addition, cervical abnormalities were found in a significant proportion of the cases.     

Comparison of the performance in detection of HPV infections between the high‐risk HPV genotyping real time PCR and the PCR‐reverse dot blot assays

A new multiplex real‐time PCR assay, the high‐risk HPV genotyping real time PCR assay (HR HPV RT‐PCR), has been developed to detect 15 high‐risk HPV types with respective viral loads. In this report, a total of 684 cervical specimens from women diagnosed with vaginitis were assessed by the HR HPV RT‐PCR and the PCR reaction and reverse dot blot (PCR‐RDB) assays, using a PCR‐sequencing method as a reference standard. A total coincidence of 97.7% between the HR HPV RT PCR and the PCR‐RDB assays was determined with a Kappa value of 0.953. The HR HPV RT PCR assay had sensitivity, specificity, and concordance rates (accuracy) of 99.7%, 99.7%, and 99.7%, respectively, as confirmed by PCR‐sequencing, while the PCR‐RDB assay had respective rates of 98.8%, 97.1%, and 98.0%. The overall rate of HPV infection, determined by PCR‐sequencing, in women diagnosed with vaginitis was 49.85%, including 36.26% of single infection and 13.6% of multiple infections. The most common infections among the 15 high‐risk HPV types in women diagnosed with vaginitis were HPV‐52, HPV‐16, and HPV‐58, with a total detection rate of 10.23%, 7.75%, and 5.85%, respectively. We conclude that the HR HPV RT PCR assay exhibits better clinical performance than the PCR‐RDB assay, and is an ideal alternative method for HPV genotyping. In addition, the HR HPV RT PCR assay provides HPV DNA viral loads, and could serve as a quantitative marker in the diagnosis and treatment of single and multiple HPV infections.     

High‐risk HPV types in lesions of the uterine cervix of female commercial sex workers in the Philippines

In order to prevent cervical cancer, vaccines against human papilloma virus types 16 (HPV‐16) and 18 (HPV‐18) have been implemented worldwide. However, the HPV types that cause cancer can differ according to geographical area and ethnicity. In this new era of the HPV vaccine, it is important to elucidate the prevalent HPV types in each area. Therefore, the prevalence of HPV infection and cervical abnormalities among 369 female commercial sex workers in the Philippines were examined. HPV L1 gene was amplified by polymerase chain reaction (PCR) using modified GP5+/6+ primers, and genotyping was performed by sequencing cloned PCR products. HPV DNA was detected in 211 (57.2%) women, among whom 46 HPV types were identified. HPV‐52 was most common and multiple‐type infection was observed in 44.5%. Among 56 women with abnormal cervical cytology (low‐ and high‐grade squamous intraepithelial lesions and adenocarcinoma in situ), HPV‐52 was most common (23.2%), followed by HPV‐16 (19.6%), ‐58 (10.7%), and ‐67 (10.7%). Only 27% of these women were positive for HPV‐16 and ‐18. Multivariate analysis revealed that HPV‐16, ‐39, ‐52, ‐67, and ‐82 were significantly associated with abnormal cytology. Repeated analysis of HPV‐52 single‐positive samples using the original GP5+/6+ PCR primers produced negative results in 57% of cases, suggesting that the prevalence of HPV‐52 infection may have been underestimated in previous studies, and the current vaccines may not be sufficient for preventing infection and the development of premalignant lesions of the cervix in women in the Philippines. J. Med. Virol. 81:545–551, 2009. © 2009 Wiley‐Liss, Inc.