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1.
子宫颈刮片中人乳头瘤病毒的基因分型 总被引:7,自引:0,他引:7
王金花 《中华实验和临床病毒学杂志》1999,(1):9-12
目的 确定不同型人乳头瘤病毒(HPV)感染的自然历程以及其持续感染在子宫颈癌发展过程中的作用。方法 应用聚合酶链反应检测荷兰155名妇女子宫刮颈片中的HPV DNA,应用线样探针分析法(LiPA)进行,包括HPV6,11,16,18,31,33,35,40,42,43,44,45,51,52,56和58的基因分型。结果 155例妇女子宫颈片中HPV DNA检出率为60%,其中在宫颈细胞学检查正常或 相似文献
2.
The PapilloCheck® assay was compared with the Digene HC2 HPV DNA assay for the detection of 13 high‐risk human papillomaviruses (HPV) in 240 samples, including 181 cervical scrapes and 59 anal scrapes. Overall, 75 (30.5%) samples were positive by the Digene HC2 HPV DNA assay: 34 (18.8%) cervical scrapes and 41 (69.5%) anal scrapes. By considering only the 13 high‐risk HPV types detected by the Digene HC2 HPV DNA assay, 66 (27.5%) samples were positive by the PapilloCheck® assay: 27 (14.9%) cervical scrapes and 39 (66.1%) anal scrapes. Concordant results between the two assays were obtained for 225 (93.8%) samples with a Kappa coefficient value of 0.85, indicating an excellent agreement. By considering all the HPV types detectable by the PapilloCheck® assay, the overall prevalence of HPV was 34.2% (82/240): 21.0% (38/181) in cervical scrapes and 74.6% (44/59) in anal scrapes. Among the samples positive by the PapilloCheck® assay, a multiple HPV infection (2–9 HPV types) was identified in 43 of 82 (52.4%) samples, including 7 of 38 (18.4%) cervical samples, and 36 of 44 (81.8%) anal samples. The prevalence of high‐risk HPV, as determined by the PapilloCheck® assay, was 17.6% (36/205) in samples with normal cytology, 83.9% (26/31) in samples with low‐grade squamous intraepithelial lesions or atypical squamous cells of undetermined significance, and 100% (4/4) in samples with high‐grade squamous intraepithelial lesions. The results obtained indicate that the PapilloCheck® assay may be considered as a reliable screening test for HPV detection and typing. J. Med. Virol. 83:1377–1382, 2011. © 2011 Wiley‐Liss, Inc. 相似文献
3.
p16INK4A, CDC6, and MCM5: predictive biomarkers in cervical preinvasive neoplasia and cervical cancer 总被引:17,自引:0,他引:17
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Murphy N Ring M Heffron CC King B Killalea AG Hughes C Martin CM McGuinness E Sheils O O'Leary JJ 《Journal of clinical pathology》2005,58(5):525-534
AIM: To analyse and compare expression patterns of three potential biomarkers-p16(INK4A), CDC6, and MCM5-and evaluate their use as predictive biomarkers in squamous and glandular cervical preinvasive neoplasia. METHODS: Immunocytochemical analysis of p16(INK4A), MCM5, and CDC6 expression was performed on 20 normal, 38 cervical intraepithelial neoplasia 1 (CIN1), 33 CIN2, 46 CIN3, 10 squamous cell carcinoma, 19 cervical glandular intraepithelial neoplasia (cGIN), and 10 adenocarcinoma samples. Staining intensity was assessed using a 0-3 scoring system. p16(INK4A), MCM5, and CDC6 expression was also examined in ThinPrep slides exhibiting mild, moderate, and severe dyskaryosis. Human papillomavirus (HPV) was detected using a modified SYBR green assay. Fluorogenic polymerase chain reaction (PCR) and solution phase PCR were used for specific HPV typing. RESULTS: All three markers showed a linear correlation between expression and grade of dysplasia. p16(INK4A) and MCM5 protein expression was upregulated in all grades of squamous and glandular dysplasia. CDC6 protein was preferentially expressed in high grade lesions and in invasive squamous cell carcinoma. CONCLUSION: p16(INK4A) expression was closely associated with high risk HPV infection-all grades of squamous and glandular cervical lesions were immunohistochemically positive. MCM5 staining intensity was independent of high risk HPV infection, highlighting its potential as a biomarker in both HPV dependent and independent cervical dysplasia. CDC6 may be a biomarker of high grade and invasive lesions of the cervix, with limited use in low grade dysplasia. p16(INK4A) was the most reliable marker of cervical dysplasia. Combinations of dysplastic biomarkers may be useful in difficult diagnostic cases. 相似文献
4.
Comparison of the SPF10-LiPA system to the Hybrid Capture 2 Assay for detection of carcinogenic human papillomavirus genotypes among 5,683 young women in Guanacaste, Costa Rica 总被引:2,自引:0,他引:2
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Safaeian M Herrero R Hildesheim A Quint W Freer E Van Doorn LJ Porras C Silva S González P Bratti MC Rodriguez AC Castle P;Costa Rican Vaccine Trial Group 《Journal of clinical microbiology》2007,45(5):1447-1454
The objective of this analysis was to compare the performance characteristics of two human papillomavirus (HPV) DNA detections assays, the Hybrid Capture 2 assay (HC2) and the SPF(10) assay, for the detection of carcinogenic HPV. Data are from the enrollment visits of women who participated in the randomized, double-blind, placebo-controlled phase III HPV16/18 Vaccine Trial in Guanacaste, Costa Rica. We compared the results of HC2 and SPF(10) testing of cervical specimens. Since the line probe assay (LiPA) detection system does not distinguish between HPV type 68 (HPV68; which is targeted by HC2) and HPV73 (which is not targeted by HC2), for SPF(10)-LiPA, we defined the carcinogenic HPV types as the 12 HC2-targeted types (types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, and 59), HPV68/73, and the HC2-cross-reactive, carcinogenic type HPV66. The kappa values and the performance characteristics for the detection of cervical abnormalities were ascertained. Paired observations were available for 5,683 sexually active, young women (median age, 21 years). The prevalence of carcinogenic HPV types was 35% (n=1,962) by HC2 and 35% (n=2,003) by SPF(10)-LiPA. There were no differences in the prevalence of carcinogenic HPV types by HC2 and SPF(10)-LiPA among women with normal, atypical squamous cells of undetermined significance and high-grade squamous intraepithelial lesion cytology. Among women with low-grade squamous intraepithelial lesion cytology, HC2 was more likely to test positive than SPF(10)-LiPA for the carcinogenic HPV types (87% and 79%, respectively; P=0.001) as a result of HC2 cross-reactivity with HPV types 40, 43, 44, 53, 54, 60, 70, and 74. The crude agreement between the two assays was 88%, with a kappa value of 0.75 (95% confidence limits, 0.73 to 0.76). We observed very good agreement between HC2 and SPF(10)-LiPA for carcinogenic HPV type detection. 相似文献
5.
Development and clinical evaluation of a highly sensitive PCR-reverse hybridization line probe assay for detection and identification of anogenital human papillomavirus. 总被引:26,自引:0,他引:26
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B Kleter L J van Doorn L Schrauwen A Molijn S Sastrowijoto J ter Schegget J Lindeman B ter Harmsel M Burger W Quint 《Journal of clinical microbiology》1999,37(8):2508-2517
Human papillomavirus (HPV) can be detected by amplification of viral DNA. A novel PCR primer set generating a short PCR fragment (SPF PCR) was used for amplification of a fragment of only 65 bp from the L1 region and permitted ultrasensitive detection of a broad spectrum of HPV genotypes. The intra- and intertypic sequence variations of the 22-bp interprimer region of this amplimer were studied. Among 238 HPV sequences from GenBank and clinical specimens, HPV genotypes were correctly identified based on the 22-bp sequence in 232 cases (97.2%). Genotype-specific probes for HPV genotypes 6, 11, 16, 18, 31, 33 to 35, 39, 40, 42 to 45, 51 to 54, 56, 58, 59, 66, 68, 70, and 74 were selected, and a reverse hybridization line probe assay (LiPA) (the INNO-LiPA HPV prototype research assay) was developed. This LiPA permits the use of amplimers generated by the SPF as well as the MY 09/11 primers. The assay was evaluated with a total of 1, 354 clinical specimens, comprising cervical scrapes (classifications ranging from normal cytology to severe dyskaryosis) and formalin-fixed, paraffin-embedded cervical carcinoma samples. LiPA results were highly concordant with sequence analysis of the SPF amplimer, genotype-specific PCR, and sequence analysis of amplimers generated by MY 09/11 primers. The sensitivity of the SPF primers was higher than that of the GP5(+)/6(+) primers over a broad range of HPV types, especially when multiple HPV genotypes were present. In conclusion, the SPF LiPA method allows extremely sensitive detection of HPV DNA as well as reliable identification of HPV genotypes in both cervical smears and paraffin-embedded materials. 相似文献
6.
da Silva Barros NK Costa MC Alves RR Villa LL Derchain SF Zeferino LC Dos Santos Carneiro MA Rabelo-Santos SH 《Journal of medical virology》2012,84(7):1143-1150
High-risk human papillomavirus (HPV) is considered the main etiological agent for cervical neoplasia. However, the presence of a single type HPV infection alone is unlikely to be sufficient to cause cervical cancer. There is epidemiologic evidence suggesting that HPV and Chlamydia trachomatis play a central role in the etiology of cervical intraepithelial neoplasia and subsequent cervical cancer. To evaluate the HPV prevalence and the seropositivity for C. trachomatis in women referred to the colposcopy clinic due to an abnormal cervical smear and to examine the effect of this association on the severity of cervical neoplasia. Following enrollment, 131 patients underwent colposcopy and biopsies when necessary. HPV DNA was detected by the polymerase chain reaction (PCR) and genotyping was performed by reverse line-blot hybridization assay. C. trachomatis seropositivity was tested by ELISA for the detection of IgG antibodies. The prevalence of HPV infection was 86.3%. Seropositivity for C. trachomatis was 26%. Thirty-one women (27.4%) were positive for C. trachomatis antibodies and HPV-DNA. The most prevalent HPV type in C. trachomatis-seropositive women were HPV 16 (51.6%) and this HPV type was present mainly in neoplasia cases. Positivity for HPV, particularly HPV types 16 and 18, and C. trachomatis seropositivity was significantly associated with a diagnosis of high grade neoplasia. Borderline significance was observed after adjustment for HPV. C. trachomatis seropositivity is associated with high grade neoplasia in women infected with HPV, mainly when the types 16 and 18 were involved. 相似文献
7.
Detection of specific types of human papillomavirus in cervical scrapes, anal scrapes, and anogenital biopsies by DNA hybridization 总被引:2,自引:0,他引:2
B R Henderson C H Thompson B R Rose Y E Cossart B J Morris 《Journal of medical virology》1987,21(4):381-393
Specific varieties of human papillomavirus (HPV) infecting the anogenital region were detected in clinical samples by use of a filter hybridization technique suitable for rapid screening of cervical and anal scrapes. In this way possibly benign types (HPV6 and HPV11) could be differentiated from types thought to be capable of malignant transformation (HPV 16 and HPV 18). Cervical or anal canal cells were applied directly to nylon filters and fixed by u.v. irradiation before hybridization with mixed viral DNA probes under both low- and high-stringency conditions. In addition, probe for the human Alu-repeated DNA sequence was used to assess the relative amount of total nucleic acids in each sample applied to the filter. HPV DNA was detected in 3 of 19 cervical scrapes from patients with no past or present history of wart virus infection or cervical dysplasia. Within a positive study group totalling 71 patients, HPV (6/11 or 16/18) was detected in cervical scrapes from 24% of 41 patients who did not have visible genital dysplasia, 30% of 27 patients with visible genital dysplasia or cervical intraepithelial neoplasia (CIN) I, and in 1 of 3 patients with past CIN II/III. In addition, HPV6/11 or 16/18 DNA was detected in anal scrapes from 3 of 6 male patients and in 85% of genital biopsies. A notably high proportion (4/6) of vaginal condylomata were positive with both the HPV6/11 and the HPV16/18 mixed viral DNA probes. Of the biopsies prepared for histopathology and positive for HPV DNA, the HPV group-specific antigen could be detected in only 60%. 相似文献
8.
Genotyping of human papillomavirus in liquid cytology cervical specimens by the PGMY line blot assay and the SPF(10) line probe assay 总被引:9,自引:0,他引:9
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van Doorn LJ Quint W Kleter B Molijn A Colau B Martin MT Kravang-In Torrez-Martinez N Peyton CL Wheeler CM 《Journal of clinical microbiology》2002,40(3):979-983
A comparison of two PCR-based human papillomavirus (HPV) DNA detection and genotyping systems (PGMY LBA and SPF(10) LiPA) was conducted in two laboratories. Both systems are based on broad-spectrum PCR for the detection of HPV DNA, followed by reverse hybridization with type-specific probes. A total of 400 selected cervical scrape specimens in PreservCyt solution (55% normal cytology, 18% atypical squamous cells of unknown significance, 14.8% low-grade squamous intraepithelial lesions [SIL], and 12.5% high-grade SIL) were tested for the presence of HPV DNA. In this selected group of specimens, the overall agreement between the two methods for the detection of any HPV DNA was high (kappa = 0.859). When the 20 common HPV genotypes identified by both methods were considered (HPV types 6, 11, 16, 18, 31, 33, 35, 39, 40, 42, 45, 51, 52, 53, 54, 56, 58, 59, 66, and 68), compatible genotype-specific results were observed in 96.5% of the samples, even when multiple HPV genotypes were present. However, for some specific HPV genotypes, there were significant differences in HPV detection by the two methods. PGMY LBA detected more HPV type 42 (P = 0.002), HPV type 56 (P = 0.039), and HPV type 59 (P < 0.001), whereas SPF(10) LiPA detected more HPV type 31 (P < 0.001) and HPV type 52 (P = 0.031). For the remaining genotypes, including HPV types 16 and 18, the results obtained by the two methods were not significantly different. In general, both genotyping methods are highly suitable for clinical and epidemiological studies. 相似文献
9.
D Riethmuller C Gay X Bertrand D Bettinger J P Schaal J P Carbillet C Lassabe P Arveux E Seilles C Mougin 《Diagnostic molecular pathology》1999,8(3):157-164
The purpose of this study was to evaluate the clinical use of the Hybrid Capture (HC)-II system for the detection of human papillomavirus (HPV) DNA to identify women at risk of progression to high grade squamous intraepithelial lesions (HGSIL) and carcinomas by differentiating low risk (LR) HPV types (6, 11, 42, 43, 44) and high/intermediate risk (HR) HPV types (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68). Five hundred and ninety-six women were enrolled in the study. Among them, 466 attended the hospital for routine cytologic screening and 130 were referred for colposcopy because of an abnormal Pap smear. The presence of HPV DNA was tested in cervical samples collected with the Digene Cervical Sampler in Digene Specimen Transport Medium (Digene Corporation, Silver Spring, MD, U.S.A.) using the HC-II assay. Results were compared with those obtained by polymerase chain reaction (PCR) using the MY09-MY11 primers followed by several hybridizations with specific probes. The overall HPV positivity was 32.9% by HC-II and 37.8% by PCR. Among cytologically normal smears, 19.5% were positive by HC-II (14.3% HR) and 25.1% by PCR. Of the atypical squamous cells of undetermined significance samples, 52.9% were positive by HC-II (41.1% HR) and 55.9% by PCR. Of the low grade SIL, 64.5% were positive by HC-II (59.4% HR) and 68.7% by PCR. The HPV positivity rate was found identical by both techniques in high grade smears (81.6%) and squamous cervical carcinomas (100%). By using PCR as the reference method, the sensitivity of HC-II was higher among women with abnormal cytology than with normal cytology (87.3% vs. 70%). Specificity was 80.8% and 97.5%, respectively. In summary, these results indicate that the HC-II method and MY-PCR identified nearly equivalent prevalences of HPV in cervical smear specimens. 相似文献
10.
Lee HP Kim SO Hwang TS Bae JM Kim SN Kim JW Hwang SY Lee HS Shin SK Cho W Hong SP 《Journal of medical virology》2011,83(3):471-482
Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry-based restriction fragment mass polymorphism (RFMP) assay was adapted to human papillomavirus (HPV) genotyping. The analytical sensitivity and the clinical utility were evaluated by testing defined HPV genome equivalents and a total of 426 specimens composed of normal cytology, atypical squamous cells of undetermined significance, low grade squamous intraepithelial lesion, high grade squamous intraepithelial lesion and invasive squamous cell carcinoma. The RFMP assay was able to detect 38.4-114.6 genomic equivalents of a wide variety of HPV types. The RFMP assay detected 34 different HPV genotypes in cervical samples of which 8% were found to be multiple-type infections. The high-risk HPV positivity rate according to the histological diagnosis was 7.9% (8/101), 31.7% (38/120), 50% (55/110), 86% (37/43), 96.2% (50/52) in normal, atypical squamous cells of undetermined significance, low grade squamous intraepithelial lesion, high grade squamous intraepithelial lesion and squamous cell carcinoma subgroups, respectively. Diagnostic sensitivities/specificities for the cervical lesions of squamous cell carcinoma and high grade squamous intraepithelial lesion or worse histology were found to be 96.2%/92.1% and 91.6%/92.1%, respectively. The sensitivity, accuracy, wide range of genotype identification and high-throughput capacity with cost-effectiveness of the test consumables make the RFMP assay suitable for mass screening and monitoring of HPV-associated cervical cancer. 相似文献
11.
p. SHEPHERD D. LUNNY R. BROOKES T. PALMER D. McCANCE† 《Scandinavian journal of immunology》1992,36(S1):69-74
The presence of human papillomavirus (HPV) types 6, 16 and 8 in cervical biopsies can be detected by an immunoperoxidase technique using type-restricted monoclonal antibodies raised against fusion proteins representing the L1 major capsid proteins of these three HPV types. In a retrospective study ( n = 54) we have used these antibodies and biotinylated DNA probes of HPV 6.16 and 18 lo detect and type HPV in formalin-fixed material from the cervix. The biopsies were classified histologically into normals, wart infections without dysplasia, cervical intraepithelial neoplasia (CIN)and squamous cell carcinomas. Antibody staining showed that 22% of all CIN was positive for HPV 16and 40% of cervical warts were positive for HPV 6, 16 and 18, There was no HPV capsid protein detected in the normals and squamous cell carcinomas using these antibodies, whereas 25% of the tumours were positive for HPV 16 by in situ hybridization. Sections of cervical warts and CIN positive for HPV types by in situ hybridization were also positive by antibody staining which suggests that both techniques are detecting replicating virus. We feel these two techniques complement each other in detection and typing of HPV in cervical biopsies from patients with active disease. 相似文献
12.
Comparative analysis of human papillomavirus infections in cervical scrapes and biopsy specimens by general SPF(10) PCR and HPV genotyping 总被引:4,自引:0,他引:4
Quint WG Scholte G van Doorn LJ Kleter B Smits PH Lindeman J 《The Journal of pathology》2001,194(1):51-58
Human papillomavirus (HPV) can be detected by DNA amplification from clinical samples. The aim of the present study was to compare the HPV status in both cervical scrape and biopsy specimens obtained from 174 patients, using the recently developed broad spectrum SPF(10) PCR-LiPA method. The detection rate of HPV in these materials was determined and the spectrum of HPV genotypes was compared. Cervical scrapes and biopsy specimens were obtained, either on the same day (group I), or with an interval of up to almost 2 years (group II, mean interval 97 days, range 1-469 days). HPV DNA was amplified by SPF(10) PCR and detected in a microtitre plate hybridization assay. Of the HPV-positive cases, the genotype was determined by reverse hybridization of the same SPF(10) amplimer on a line probe assay (LiPA), discriminating between HPV genotypes 6, 11, 16, 18, 31, 33-35, 39, 40, 42-45, 51-54, 56, 58, 59, 66, 68, 70, and 74. The results showed that the detection rate and the spectrum of HPV genotypes in cervical scrapes and the corresponding biopsy specimens were highly comparable in both patient groups, even when multiple genotypes were present. In both groups, multiple HPV genotypes were more frequently detected in cervical scrapes than in the corresponding biopsy specimens. In conclusion, HPV infection can be diagnosed in cervical scrapes and biopsy specimens using the SPF(10) PCR-LiPA system. Analysis of cervical scrapes accurately reflects the spectrum of HPV genotypes in the patient's cervical region, even with a sampling interval between the cervical scrape and the biopsy specimen. 相似文献
13.
Detection and typing of human papillomavirus DNA by PCR using consensus primers in various cervical lesions of Korean women 总被引:2,自引:0,他引:2
Hwang T 《Journal of Korean medical science》1999,14(6):593-599
The association between cervical cancers and human papillomavirus (HPV) is now well established. To estimate the extent of infection with common HPVs among Korean women, we have examined 224 cervical scrapes of various cervical lesions. Detection and typing of HPVs were done by polymerase chain reaction (PCR) using consensus primers followed by restriction enzyme digestion and PCR using type-specific primers. The prevalence of total HPV infection in patients with cervical intraepithelial neoplasia (CIN) and cervical cancer were significantly higher than those in healthy women and patients with atypical squamous cells of undetermined significance (ASCUS). HPV typing in 41 invasive carcinomas of the cervix revealed the prevalence of HPV 16 in 15 cases, followed by HPV 58, 18, 33, 31, 52 and 35. The distribution pattern of HPV types in CIN were not much different from carcinomas. HPV types except HPV 18 had a tendency to show higher prevalence in high-grade squamous intraepithelial lesion (HSIL) than low-grade squamous intraepithelial lesions (LSIL), however, HPV 18 was detected in LSIL but not in HSIL. HPV 18 tended to have the worse clinical stage, although it was not statistically significant. These findings suggest the importance of HPV typing other than HPV 16 and 18 and a different clinicopathologic significance of HPV 18. 相似文献
14.
Carcopino X Henry M Benmoura D Fallabregues AS Richet H Boubli L Tamalet C 《Journal of medical virology》2006,78(8):1131-1140
It has been recognized that human papillomavirus infection is the major causal factor for high-grade cervical lesions. The aim of the study was to evaluate the relationship between HPV 16 and 18 viral loads and cervical status in different age strata. A duplex real time PCR method was devised to determine HPV 16 and 18 viral load per million of human cells using an in house plasmidic construct as a standard of quantification. The 151 cervical scrapes were collected before colposcopic examination from either abnormal cervico-vaginal smear (group 1, 97 patients) or from post treatment clinical follow-up (group 2, 54 patients). In women aged 30-40, the HPV16 viral loads were significantly higher in high-grade squamous intraepithelial lesion than in low-grade squamous intraepithelial lesion in both groups and HPV18 in group 1. In women aged 20-30 of group 1, high HPV viral load was associated in few cases with high-grade squamous intraepithelial lesion or low-grade squamous intraepithelial lesion, and surprisingly in some patients with normal cervix. HPV 16 and 18 viral loads are related to the severity of cervical lesion, and may be useful in the clinical management of cervical lesions. A specific follow-up may be useful for those with high viral load despite normal cervix. 相似文献
15.
Mendoza LP Arbiza J Paez M Kasamatsu E Castro A Giménez G Basiletti J Gonzalez J Mongelós P Picconi MA 《Journal of medical virology》2011,83(8):1351-1357
The incidence of cervical cancer in Paraguay is among the highest in the world. This study aimed to determine the distribution of human papillomavirus (HPV) genotypes in Paraguayan women, according to the severity of the cervical lesion. This cross-sectional study included 207 women without a squamous intraepithelial lesion, 164 with low-grade squamous intraepithelial lesions, 74 with high-grade squamous intraepithelial lesions, and 41 with cervical cancer. Type-specific HPV was determined by the polymerase chain reaction with MY9/11 L1 and GP5+/GP6+ L1 primers, followed by restriction fragment length polymorphism and reverse line blotting hybridization, respectively. In total, 12 high-risk and 24 low-risk HPVs types were detected. HPV 16 was the most prevalent, followed by HPV 18 in cervical cancer (14.6%), HPV 31 in high-grade squamous intraepithelial lesions (14.9%), HPVs 58/42 in low-grade squamous intraepithelial lesions (9.1% each), and HPVs 31/58 (2.4% each) in women without squamous intraepithelial lesions. Among 285 positive samples, 24.2% harbored multiple HPV types, being this more prevalent in women with squamous intraepithelial lesions (30.8% in low-grade squamous intraepithelial lesions, 22.5% in high-grade squamous intraepithelial lesions, and 22.0% in cervical cancer) than in women without lesions (9.3%). The higher prevalence of HPV 16 and other high-risk HPVs in women both with and without cervical lesions may explain the high incidence of cervical cancer in Paraguay. This information may be of importance for local decision makers to improve prevention strategies. In addition, these results may be useful as baseline pre-vaccination data for a future virological surveillance in Paraguay. 相似文献
16.
Use of anticontamination primers in the polymerase chain reaction for the detection of human papilloma virus genotypes in cervical scrapes and biopsies 总被引:13,自引:0,他引:13
A J van den Brule E C Claas M du Maine W J Melchers T Helmerhorst W G Quint J Lindeman C J Meijer J M Walboomers 《Journal of medical virology》1989,29(1):20-27
A reliable application of the polymerase chain reaction (PCR) for detection of the human papilloma virus (HPV) genotypes in cervical smears and biopsies was developed. Primers flanking the HPV cloning site were used to avoid detection of cloned HPV plasmids. These anticontamination primers were used for the specific detection of HPV 6, 11, 16, 18, and 33 in cervical scrapes that had been tested previously for HPV with a combined modified filter in situ hybridization (modified FISH) and dot blotting procedure. The PCR appeared to be superior. Two groups of women were screened for HPV genotypes. Group A consisted of women belonging to a regularly screened population, and group B contained women attending a gynaecological clinic. It appeared that the overall prevalence of HPV in cytologically normal scrapes in the first group was 6%, whereas in the second group 12% was found. In scrapes with cytological dysplasia, the prevalence of HPV in group A and B was approximately 40% and 60%, respectively. HPV 16 was present predominantly. In biopsies of squamous cell carcinomas of the cervix uteri, an HPV prevalence rate of 90% was found, all of which contained only HPV 16 and 18. These data indicate an important role for HPV detection in the screening of cervical scrapes to identify women with an increased risk of cervical cancer. 相似文献
17.
Increased detection rate of human papillomavirus in cervical scrapes by the polymerase chain reaction as compared to modified FISH and southern-blot analysis 总被引:17,自引:0,他引:17
W Melchers A van den Brule J Walboomers M de Bruin M Burger P Herbrink C Meijer J Lindeman W Quint 《Journal of medical virology》1989,27(4):329-335
Cervical scrapes from 80 women with a positive cytology result were tested for the presence of human papillomavirus (HPV) using the polymerase chain reaction (PCR) and compared to the results obtained with the modified filter in situ hybridisation (FISH) and the Southern-blot techniques. The sensitivity of the modified FISH and the Southern-blot was similar, and HPV was detected in 46% of the patients. The sensitivity of the PCR appeared to be higher, and HPV was detected in 70% of the patients. HPV-DNA could be detected in 46 of the 68 patients with mild dysplasia, in 6 of the 8 patients with severe dysplasia, and in all 4 patients with carcinoma in situ. In 18 patients (21%) more than one HPV type could be detected by the PCR. The control group consisted of 100 women involved in a triennial checkup programme, who had normal smear results and no history of cervical lesions. HPV was detected in 5% of the women by the PCR. The PCR technique detected HPV in a high proportion of the cervical scrapes from women with a positive cytology result. These results give further evidence for an important role of HPV in the pathogenesis of cervical cancer. 相似文献
18.
Rachel L. Winer Tiffany G. Harris Long Fu Xi Kathrin U. Jansen James P. Hughes Qinghua Feng Carolee Welebob Jesse Ho Shu‐Kuang Lee Joseph J. Carter Denise A. Galloway Nancy B. Kiviat Laura A. Koutsky 《Journal of medical virology》2009,81(4):713-721
Human papillomavirus (HPV) RNA levels may be a more sensitive early indicator of predisposition to carcinogenesis than DNA levels. We evaluated whether levels of HPV‐16 and HPV‐18 DNA and messenger RNA (mRNA) in newly detected infections are associated with cervical lesion development. Female university students were recruited from 1990 to 2004. Cervical samples for HPV DNA, HPV mRNA, and Papanicolaou testing were collected tri‐annually, and women were referred for colposcopically directed biopsy when indicated. Quantitative real‐time polymerase chain reaction of L1 and E7 DNA and E7 mRNA was performed on samples from women with HPV‐16 and HPV‐18 infections that were incidently detected by consensus PCR. Adjusting for other HPV types, increasing E7 cervical HPV‐16 mRNA levels at the time of incident HPV‐16 DNA detection were associated with an increased risk of cervical intraepithelial neoplasia grade 2–3 (HR per 1 log10 increase in mRNA = 6.36, 95% CI = 2.00–20.23). Increasing HPV‐16 mRNA levels were also associated with an increased risk of cervical squamous intraepithelial lesions; the risk was highest at the incident positive visit and decreased over time. Neither HPV‐16 E7 DNA levels nor HPV‐18 E7 DNA nor mRNA levels were significantly associated with cervical lesion development. Report of >1 new partner in the past 8 months (relative to no new partners) was associated with increased HPV mRNA (viral level ratio [VLR] = 10.05, 95% CI = 1.09–92.56) and increased HPV DNA (VLR = 16.80, 95% CI = 1.46–193.01). In newly detected HPV‐16 infections, increasing levels of E7 mRNA appear to be associated with an increased risk of developing cervical pre‐cancer. J. Med. Virol. 81:713–721, 2009 © 2009 Wiley‐Liss, Inc. 相似文献
19.
High-grade cervical intraepithelial lesions (HGCINs) are easily diagnosed by established histologic criteria. However, we encountered problematic cases that are difficult to diagnose because features intermediate between dysplasia and metaplasia are present. p16 and Ki-67 immunostains proved HGCIN in these difficult and unusual cases. Because these are unusual cases of cervical dysplasia, we decided to type the human papillomavirus (HPV) using the Invader HPV test with analyte-specific reagents developed by Third Wave Technologies (Madison, WI, USA) (a new HPV screening assay applicable to tissue and amenable to rapid, sensitive, and specific detection of 14 high- to intermediate-risk HPV types) and a panel of immunostains. Results of these difficult cases are compared with classic HGCIN cases. We searched our pathology files over a period of 16 months for high-grade squamous intraepithelial lesion, cervical intraepithelial neoplasia II, cervical intraepithelial neoplasia III, and p16. To identify cases of difficult HGCIN with features intermediate between dysplasia and metaplasia, we reviewed all surgical cases of HGCIN that required p16 and Ki-67 diagnosis confirmation. Cases of interest were also stained with ProExC. Human papillomavirus screening and HPV 16/18 typing were performed by the Invader assays as described previously. Ten cases of classic HGCIN were easily diagnosed by hypercellularity, significant atypia, mitotic figures, and diffuse staining by p16, Ki67 and ProExC. The Invader assay identified HPV 16 (A9 positive/HPV16 positive) in 7 of 10 cases; the 3 others were A7 positive/not HPV18 (1) and A9 positive/not HPV16 (2). Eight cases of difficult HGCIN were identified. These showed only mild-to-moderate cellularity, a lack of significant atypia, absent-to-rare mitotic figures, and diffuse staining by p16, Ki-67, and ProExC. Human papillomavirus DNA was detected in 5 of 8 cases: only 1 was A9 positive/HPV16 positive, 1 was A5/A6 positive, 1 was A7 positive/not HPV18, and 2 were A9 positive/not HPV16. Three remaining cases demonstrated sufficient DNA to be analyzed by the Invader assay, but results were negative. This is a poorly recognized unusual group of cervical HGCIN with features intermediate between dysplasia and metaplasia that is easily confused by histologic examination. Immunostains prove the high-grade nature of these lesions, and Invader assay demonstrates association with HPV types other than 16/18 (ie, other HPV types detected by Invader assay). In this study, we present an unusual group of cases of high-grade dysplasia, not recognized by hematoxylin and eosin but identified by Ki67 and P16. It is very important to emphasize that this unusual group of high-grade dysplasias is associated with high-risk HPV but with types other than 16/18. 相似文献
20.
Human papillomavirus detection by hybrid capture and its possible clinical use. 总被引:3,自引:4,他引:3
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AIMS--To determine the sensitivity of the hybrid capture method for human papillomavirus (HPV) detection and potential clinical uses as a screening method for the identification of cervical intraepithelial neoplasia. METHODS--The presence of oncogenic types of HPV was tested for in samples taken from the cervix at colposcopy, and compared with detection by polymerase chain reaction (PCR) in 60 patients. Both sets of results were corrected with the pathology determined by biopsy and smear cytology. RESULTS--Hybrid capture detection showed 86% agreement with PCR. Eighty three percent of CIN 3 lesions, 62% of CIN 2, 59% of CIN 1 and 21% of normal controls were positive for oncogenic HPV types. CONCLUSION--The hybrid capture detection method is reliable, sensitive, and easy to use. The addition of HPV testing to cytological screening would detect a greater proportion of cervical dysplasia with a higher false positive rate. 相似文献