HPV6，11型在女性生殖器尖锐湿疣和乳头瘤样增生中的分布

从３４２名上海地区患者的外阴、阴道和宫颈的病变部，采集４８７份活检标本。５种病理诊断类型，经斑点杂交法检测，ＨＰＶ６、１１ＤＮＡ总阳性率：尖锐湿疣８５％、乳头瘤样增生１０．１０％、慢性宫颈炎７．１４％、鳞状上皮增生５％、生殖器正常粘膜２．６５％。５１例尖锐湿疣和１０例乳头瘤样增生，ＨＰＶ６、１１和６＋１１感染阳性率分别为：１５．６９％、２５．４９％、５８．８２％和２０％、３０％、５０％。Ｓｏｕｔｈｅｒｎ印迹转移杂交与斑点杂交的符合率，在尖锐湿疣中，ＨＰＶ６、１１和６＋１１分别为７５％、９２．３％、８６．７％；在乳头瘤样增生中，分别为１００％、３３．３％、６０％。ＡＢＣ法检测阳性率仅为６１．６７％和０％。本调查结果证实，上海地区女性尖锐湿疣和乳头瘤样增生与ＨＰＶ６、１１型感染密切相关，且以ＨＰＶ两种型别（ＨＰＶ６＋１１）混合感染为主。     

Detection of multiple types of human papillomavirus in a giant condyloma from a grafted patient. Analysis by immunohistochemistry, in situ hybridisation, Southern blot and polymerase chain reaction.

Immunosuppressed patients such as transplant recipients are known to develop multiple lesions suggestive of human papillomavirus (HPV) infection. A giant anal condyloma was obtained from a transplant patient; several fragments taken from different areas were examined for the presence of HPV DNA using in situ hybridisation, polymerase chain reaction (PCR) and Southern blot. Typical koilocytes were seen in routinely stained tissue sections, suggesting an HPV infection; furthermore, group specific HPV antigen was detected in one of four frozen fragments. Different results were obtained by in situ hybridisation according to the fragment tested. HPV types 6/11 were detected in each of the five fragments, frozen or fixed in Bouin's or formalin solutions. However, the number of HPV DNA positive cells and the intensity of the reaction greatly varied with the specimen. HPV 16 and 18 probes also reacted positively with the sample fixed in formalin; a stronger signal was observed with HPV 18 in one large focus than with HPV 16. HPV type 5 was detected in a few isolated cells of two frozen fragments. With the Southern blot technique, the profile of an HPV 6/11 was seen only in one of two frozen fragments; in this case, the bands were intense. A slight positive reaction was also obtained in one frozen fragment with HPV 16 probe. Four frozen fragments were analyzed with PCR: HPV 6/11 was detected in each fragment; HPV 18 was detected in the four samples but with different intensities; HPV types 5 and 16 did not show any positive signal. In conclusion, the lesion is an example of infection with several HPV types, demonstrated by three different techniques. This suggests the need for careful dermatological or colposcopic follow-up of transplant recipients, in order to prevent possible malignant transformation of anogenital lesions.     

食管鳞状上皮细胞癌及癌旁组织中HPV11 L1基因克隆及序列比较

目的 克隆食管癌组织和癌旁组织中HPV11型主要衣壳蛋白L1基因,并比较两个序列间的同源性。方法 以食管癌组织和癌旁组织纯化的总DNA为模板,根据HPV11型L1基因的保守区分段设计引物。分两段扩增HPV L1基因,克隆入质粒载体中,pGEM-3Zf(-)以双脱氧法双向测定目的片段的序列,并按接出该片段的全序列,比较两个序列间的同源性,以及与已知基因序列的差异性。结果 从4例食管癌患者食管癌组织和癌旁组织中,分别克隆到1株HPV11型L1的编码序列M1和M2,两序列间的同源性极高,仅在个别位点存在差异。与GenBank中登录的HPV序列NC001525．1（HPV－11）、M14119．1（HPV－11）和AF217526．1（HPV－11）相比较大部分相同。结论 成功地构建了HPV11型L1基因上游片段pGEM－3Gf(-)和下游片段pGEM-3Zf(-)的重组克隆,为深入研究HPV的感染与食管癌发病机制的关系奠定了基础。     

Detection of infection by human papillomavirus in genital condylomata. A comparison study using immunocytochemistry and in situ nucleic acid hybridization

One hundred eighty exophytic genital lesions clinically suspicious for infection by human papillomavirus (HPV) were analyzed by hematoxylin and eosin light microscopy, immunocytochemistry for HPV capsid antigen; and in situ nucleic acid hybridization for HPV messenger RNA (mRNA). Of 96 cases morphologically consistent with infection by HPV, 53% were antigen-positive, and 83% were mRNA positive (P less than 0.01). Of 55 cases suggestive but not diagnostic of HPV infection, 13% were antigen-positive and 26% were mRNA positive. Negative results were obtained in all lesions not believed to be indicative of HPV infection by morphologic criteria. In mRNA positive diagnostic cases, two thirds were of HPV type 6 and one third were HPV type 11. Two cases of coinfection with HPV types 6 and 16 were found. The study concludes that in situ hybridization for HPV mRNA is a more sensitive indicator of HPV infection, and in addition, provides HPV type, which may have prognostic significance.     

Human papillomavirus detection by hybrid capture in paired cervicovaginal lavage and cervical biopsy specimens

Infection of the uterine cervix with human papillomavirus (HPV) is associated with dysplastic lesions that may progress to malignancy. Certain HPV types are associated with higher risk of cervical cancer than other genital HPVs. The goal of this study was to determine if cells obtained by cervicovaginal lavage contain similar HPV types as paired cervical biopsy in women referred because of abnormal cervical cytology. Thirty-four paired lavage and biopsy samples were analyzed for HPV DNA by hybrid capture, using “low risk” (HPV types 6. 11, and related types and “high risk” group (HPV types 16, 18, and related types) HPV. HPV was detected in 24 lavage samples and 18 biopsies. High risk types were predominant. In 14 of 18 HPV-positive biopsies, the paired lavage was also positive for the same HPV group. Four biopsies were HPV-positive at low levels, and the paired lavage was HPV-negative. The mean viral copy numbers of the biopsies from patients with positive and negative lavage samples were 2.7 and 0.1, respectively (P = .02). Ten low level HPV infections were detected by lavage that were not detected by biopsy. HPV detection by hybrid capture in cells obtained by cervicovaginal lavage reflects the results of HPV testing in cervical biopsies. © 1996 Wiley-Liss, Inc.     

REBA HPV-ID® for efficient genotyping of human papillomavirus in clinical samples from Korean patients

This study was conducted to evaluate the overall performance of a reverse blot hybridization‐based assay, REBA HPV‐ID® (Molecules and Diagnostics, Wonju, Korea) for genotyping human papillomaviruses (HPV). HPV Genotyping on 356 specimens examined cytologically was performed using the REBA HPV‐ID®, and its results were compared with those obtained using the MyHPV DNA Chip® (Mygene, Seoul, Korea), DNA chip‐based HPV genotyping assay. The results from this study showed that the positivity rate of the REBA HPV‐ID® for abnormal cytological samples was higher (80.9%) than that of the MyHPV DNA chip (69.8%). In addition, the REBA HPV‐ID® positivity rate with normal cytological samples was higher (64.4%) than that obtained using DNA chips (34.4%). Subsequently, sequence analysis was performed with specimens that generated conflicting test results. Sequence analysis confirmed that the specimens which were positive by REBA HPV‐ID® did indeed contain HPV sequences. The results of this study suggest that the REBA HPV‐ID® is a sensitive test for genotyping HPV of clinical specimens. J. Med. Virol. 84: 1248–1253, 2012. © 2012 Wiley Periodicals, Inc.     

Demonstration of human papillomavirus types in paraffin processed tissue from human ano-genital lesions by in-situ DNA hybridisation

A sensitive in situ hybridization technique for the demonstration of human papillomavirus (HPV) employing a biotin-streptavidin polyalkaline phosphatase complex has been successfully applied to formalin-fixed, paraffin processed tissue obtained from a selected series of patients with ano-genital lesions. Benign condylomata from males and females showed the presence of HPV 6 and 11. Two cases of vulval intraepithelial neoplasia showed HPV 16. Four cases of squamous carcinoma of the anal canal also showed HPV 16 in the tumour or in the adjacent pre-invasive neoplastic epithelium. A case of malignant transformation in a cervical condyloma was associated with HPV 6 and 11. This technique permits the retrospective evaluation of routinely processed material thus widening the investigative spectrum for HPV.     

Real-time duplex PCR for simultaneous HPV 16 and HPV 18 DNA quantitation

HPV 16 and HPV 18 are responsible for more than 75% of cervical cancers and high HPV 16 loads are associated with both prevalent and incident lesions. The objective of the present study was to develop a method allowing the detection and quantitation of HPV 16 and 18 DNA to improve future strategies for cervical cancer screening. A duplex real-time PCR allowing the simultaneous quantitation of both HPV 16 and HPV 18 was carried out. Mixes of HPV 16 and HPV 18 whole genome plasmids were prepared to test a wide range of viral DNA concentrations. The values obtained for each mix of plasmids with the simplex and the duplex PCR were very close to the theoretical values except when a HPV type represented only 1:1000 genome equivalent or lower than the concurrent type. Cervical samples harboring HPV 16, HPV 18 or both types were tested by comparing the results with simplex and duplex real-time PCR assays. HPV 16 and HPV 18 genome titers were similar with the two assays. In conclusion, the real-time duplex PCR proved to be robust for HPV 16 and HPV 18 DNA quantitation.     

"HPV SCORE", A SCORING SYSTEM FOR HISTOLOGICAL DIAGNOSIS OF HUMAN PAPILLOMAVIRUS INFECTION IN DYSPLASIA OF THE UTERINE CERVIX

We devised a scoring system, "HPV score", for histological diagnosis of human papillomavirus (HPV) infection in dysplasia of the uterine cervix. Four hundred and sixty cases of cervical dysplasia were screened for the presence of HPV infection using this system, and 116 cases (25%) were judged to be HPV infection. The results showed good correlation with those obtained using the immunoperoxldase (IMPO) method, but 42 of the 116 cases (36%) were negative for HPV antigen by the IMPO method. Severe stromal inflammation was noted in about half of such cases. Among the four histological types of cervical condyloma, the flat type was most common among the cases with HPV infection judged according to HPV score. It was concluded that this scoring system is simple and accurate for the detection of HPV infection in cervical lesions.     

"HPV score", a scoring system for histological diagnosis of human papillomavirus infection in dysplasia of the uterine cervix

We devised a scoring system, "HPV score", for histological diagnosis of human papillomavirus (HPV) infection in dysplasia of the uterine cervix. Four hundred and sixty cases of cervical dysplasia were screened for the presence of HPV infection using this system, and 116 cases (25%) were judged to be HPV infection. The results showed good correlation with those obtained using the immunoperoxidase (IMPO) method, but 42 of the 116 cases (36%) were negative for HPV antigen by the IMPO method. Severe stromal inflammation was noted in about half of such cases. Among the four histological types of cervical condyloma, the flat type was most common among the cases with HPV infection judged according to HPV score. It was concluded that this scoring system is simple and accurate for the detection of HPV infection in cervical lesions.     

A growth model of human papillomavirus type 16 designed from cellular automata and agent-based models

ObjectiveThis paper presents a conceptual model that is developed upon a characterization of human papillomavirus type 16 (HPV16) which is used to build a simulation prototype of the HPV16 growth process.MethodologyThe human papillomavirus type 16 is the principal virus detected in invasive lesions of cervical cancer, and associated with the greater persistence and prevalence in pre-malignant and malignant lesions. The probability of acquiring an infection with HPV16 is extremely high in sexually active individuals. However, an HPV16 infection can disappear after becoming a histological confirmed case. According to the characterization of HPV16 proposed in this paper, cells as compared to a society behaves as a complex system, i.e., cells behave in a cooperative manner, following a set of rules defined by local interactions among them. Such complex system is defined by combining a cellular automaton and agent-based models. In this way, the behavior of the HPV16 is simulated by allowing the cellular automaton to follow such parameterized behavior rules.ResultsBoth cross-sectional and prospective studies indicate that HPV16 infection persistence increase the risk of high-grade CIN, as observed in the results provided by the growth simulation model of HPV16. The average growth rate extrapolated over 52 weeks (12 months) and calculated by the model showed a 37.87% growth for CIN1, 35.53% for CIN2 and 16.92% for CIN3. Remarkably, these results are similar to the results obtained and reported by clinical studies. For example, the results obtained using the proposed model for CIN2 and the results obtained by Östör [36], have a differential of 0.53 percentage points while have a differential of 2.23 percentage points with the results obtained by Insinga et al. [51]. Also, for the CIN3, the results obtained using the proposed model, have a differential of 2.92 percentage points with the Insinga et al. [52], results.ConclusionThrough the specification of parameterized behavior rules for HPV16 that are simulated under the combined technique of cellular automata and agent-based models, the HPV life cycle can be simulated allowing for observations at different stages. The proposed model then can be used as a support tool in the investigation of HPV16, in particular (as part of our future work) to develop drugs as agents in the control of the HPV16 disease.     

Comparison of the Novel Human Papillomavirus 4 Auto-capillary Electrophoresis Test with the Hybrid Capture 2 Assay and with the PCR HPV Typing Set Test in the Detection of High-Risk HPV Including HPV 16 and 18 Genotypes in Cervical Specimens

The aim of this study was to compare the novel human papillomavirus (HPV) detection method, the HPV 4 Auto-capillary Electrophoresis (ACE) test with the hybrid capture (HC) 2 assay for the detection of high-risk HPVs. In addition, we compared the HPV 4 ACE test with the polymerase chain reaction HPV Typing Set test for the detection of HPV 16 and HPV 18 genotypes. One hundred ninety-nine cervical swab samples obtained from women with previous abnormal Pap smears were subjected to testing with the three HPV tests. The HPV 4 ACE test and the HC 2 assay showed substantial agreement for detection of high-risk HPVs (85.4%, kappa=0.71). The HPV 4 ACE test also showed substantial agreement with the PCR HPV Typing Set test in the detection of HPV 16 and HP V 18 genotypes (89.9%, kappa=0.65). In correlation with cytologic results, the sensitivities and specificities of the HPV 4 ACE test and HC 2 assay were 92.9% vs. 92.9% and 48.1% vs. 50.8%, respectively, when high-grade squamous intraepithelial lesions were regarded as abnormal cytologies. The novel HPV 4 ACE test is a valuable tool for the detection of high-risk HPVs and for genotyping of HPV 16 and HPV 18.     

Use of PGMY primers in L1 consensus PCR improves detection of human papillomavirus DNA in genital samples

The novel PGMY L1 consensus primer pair is more sensitive than the MY09 and MY11 primer mix for detection and typing with PCR of human papillomavirus (HPV) DNA in genital specimens. We assessed the diagnostic yield of PGMY primers for the detection and typing of HPV by comparing the results obtained with PGMY09/PGMY11 and MY09/MY11/HMB01 on 299 genital samples. Amplicons generated with PGMY primers were typed with the line blot assay (PGMY-line blot), while HPV amplicons obtained with the degenerate primer pool MY09/MY11/HMB01 were detected with type-specific radiolabeled probes in a dot blot assay (standard consensus PCR test). Cervicovaginal lavage samples (N = 272) and cervical scrape samples (N = 27) were tested in parallel with both PCR tests. The PGMY-line blot test detected the presence of HPV DNA more frequently than the standard consensus PCR assay. The concordance for HPV typing between the two assays was 84.3% (214 of 255 samples), for a good kappa value of 0.69. Of the 177 samples containing HPV DNA by at least one method, 40 samples contained at least one HPV type detected only with PGMY-line blot, whereas positivity exclusively with the standard consensus PCR test was found for only 7 samples (P < 0.001). HPV types 45 and 52 were especially more frequently detected with PGMY than MY primers. However, most HPV types were better amplified with PGMY primers, including HPV-16. Samples with discordant results between the two PCR assays more frequently contained multiple HPV types. Studies using PGMY instead of MY primers have the potential to report higher detection rates of HPV infection not only for newer HPV types but also for well-known genital types.     

Detection of human papillomavirus type 16 DNA in carcinomas of the palatine tonsil.

Twenty eight tonsillar carcinomas of various histological types were investigated for the presence of Epstein-Barr virus (EBV), cytomegalovirus (CMV), and human papillomavirus (HPV) types 6, 11, and 16 by in situ hybridisation using highly stringent procedures. In six cases an autoradiographic signal was obtained in the tumour cell nuclei with the HPV type 16 specific probe. No signal was obtained with any of the other probes. Immunohistochemical investigations with mouse monoclonal antibodies directed against the L1 protein of HPV type and a rabbit antiserum that detects common protein determinants of HPV gave negative results, thus indicating latent infection. Furthermore, a series of tonsils from controls with comparable age distribution was negative by both in situ hybridisation and immunohistology. These results indicate a possible role for HPV 16 in the aetiology of a proportion of tonsillar carcinomas.     

金华地区女性感染人乳头瘤病毒基因类型分析

目的：了解金华地区各年龄段与不同生殖道疾病女性感染人乳头瘤病毒（humanpa pilloma virus,HPV）基因类型分布状况,为金华地区HPV分子流行病学研究提供依据。方法：利用反向杂交技术分别对2006年1月～2007年12月来医院就诊的236例可疑患者进行23种HPV基因亚型检测,其中包括18种高危亚型（HPV16、18、31、33、35、39、45、51、52、53、56、58、59、66、68、73、83、MM4）及5种低危亚型（HPV6、11、42、43、44）。结果：HPV阳性检出率为13.6%（32/236）,其中高、低危型检出率分别为13.1%（31/236）与3.4%（8/236）,高危型中主要为HPV16、18、58感染,低危型主要为HPV6、11感染。不同生殖道疾病[尖锐湿疣、宫颈上皮内瘤变（CIN）]与正常对照组比较差异有显著性意义（P〈0.05）。（20～30）岁,（31～40）岁,（41～50）岁和〉50岁各年龄组中HPV阳性的比率为33.3%,38.5%,2.31%和5.1%。结论：HPV6、11、16、18、58是金华地区HPV感染的主要型别,（20～50）岁为感染的高峰人群,对HPV阳性者进行定期的跟踪,是防治尖锐湿疣与宫颈癌的重点。     

机会性筛查人群中高危型人乳头瘤病毒与宫颈病变相关性的分析

目的 研究第二代杂交捕获法(HC2)高危型人乳头瘤病毒(HR-HPV)检测阳性样品中HPV与宫颈病变的相关性.方法 随机抽取HC2阳性患者602名(344名行细胞学检查),采用PCR-反向点杂交(RDB)法对HPV分型并分析HPV型别与细胞学的相关性.结果 602例患者PCR阳性率94.5%;HPV分型检出率依次为HPV16、52、58、56、39等;HC2检测谱外单独感染24例;HPV16、HPV16/18与ASCUS、LSIL、HSIL有明显相关性.结论 机会性筛查人群中最常见HPV型别依次为HPV16、52、58;HC2检测的假阳性率为4.0%;HPV16与宫颈病变程度相关.     

Antibody response for L1, E6 and E7 HPV 16 and HPV 18 antigens in Tunisian women with cervical cancer and controls

Results obtained in the present work indicated that the Luminex assay is more sensitive than ELISA. The reactivity to the early antigens E6 and E7 was 37% versus 42% for HPV 16 and 21% versus 20% for HPV 18 among cervical cancer cases using ELISA. However, these ratios were 44% and 61%, respectively, for E6 and E7 HPV 16 versus 28% and 21% for E6 and E7 HPV 18 when using the Luminex technique. Data also indicated that HPV 16 and HPV 18 showed distinct profiles for the different antigens tested. Finally, the differences in antibody responses between cervical cancer cases and benign cases toward the different antigens were significant.     

机会性筛查人群中高危型人乳头瘤病毒与宫颈病变相关性的分析

目的研究第二代杂交捕获法（HC2）高危型人乳头瘤病毒（HR—HPV）检测阳性样品中HPV与宫颈病变的相关性。方法随机抽取HC2阳性患者602名（344名行细胞学检查）,采用PCR一反向点杂交（RDB）法对HPV分型并分析HPV型别与细胞学的相关性。结果602例患者PCR阳性率94．5％;HPV分型检出率依次为HPVl6、52、58、56、39等;HC2检测谱外单独感染24例;HPVl6、HPVl6／18与ASCUS、LSIL、HSIL有明显相关性。结论机会性筛查人群中最常见HPV型别依次为HPVl6、52、58;HC2检测的假阳性率为4．0％;HPVl6与宫颈病变程度相关。     

机会性筛查人群中高危型人乳头瘤病毒与宫颈病变相关性的分析

目的 研究第二代杂交捕获法(HC2)高危型人乳头瘤病毒(HR-HPV)检测阳性样品中HPV与宫颈病变的相关性.方法 随机抽取HC2阳性患者602名(344名行细胞学检查),采用PCR-反向点杂交(RDB)法对HPV分型并分析HPV型别与细胞学的相关性.结果 602例患者PCR阳性率94.5%;HPV分型检出率依次为HPV16、52、58、56、39等;HC2检测谱外单独感染24例;HPV16、HPV16/18与ASCUS、LSIL、HSIL有明显相关性.结论 机会性筛查人群中最常见HPV型别依次为HPV16、52、58;HC2检测的假阳性率为4.0%;HPV16与宫颈病变程度相关.     

Comparison of the Cobas 4800 Human Papillomavirus test against a combination of the Amplicor Human Papillomavirus and the Linear Array tests for detection of HPV types 16 and 18 in cervical samples

The greater prevalence of human papillomavirus (HPV) types 16 and 18 compared to the other high-risk HPV types of cervical cancer led to the development of clinical tests that detect both types separately from other genotypes. One method is the Roche Cobas 4800 HPV test, which is based on a real-time PCR. The aim of this study was to evaluate the performance of the Cobas 4800 HPV test for detecting genotypes 16 and 18 by comparing the results with those obtained in a combination of the Roche Amplicor HPV assay and the Roche Linear Array (LA) HPV genotyping assay. Excellent concordance was found between both methods (92.7%, kappa value=0.872). The Cobas 4800 HPV test could be used as a single test for identifying HPV types 16 and 18 directly from clinical specimens.