用质量控制狭缝式杂交法（Coslot）检测妇女宫颈上皮中人乳头瘤病毒（HPV）DNA

本文报道一种能对ＨＰＶ进行较为准确分型的刚Ａ杂交程序和方法。它具有如下特点：１．由于简化了实验程序,标本消耗减少。因此该方法尤其适用于小组织活检标本（１－２ｍｇ）中ＤＮＡ同源序列检测;２．敏感性高。由于标本消耗减少,使ＨＰＶ检出率大为提高,与斑点杂交技术相比,敏感性提高１．６倍;３．型别特异性强。由于改变了实验条件及在实验过程中实行严格的质量控制,明显减少了不同ＨＰＶ型别的交叉反应,甚至在部分出现交叉反应的标本中,仍可通过强度对比来最后判定ＨＰＶ感染型别。应用该技术对１４３份宫颈炎标本、１７份宫颈不典型增生标本及２５份宫颈癌标本进行了ＨＰＶ１６型检测,阳性率分别为５０．０５％（７３／１４３）、５７％（２７／４７）及７６％（１９／２５）,并与Ｄｏｔｂｌｏｔ方法进行了比较。该方法在ＨＰＶ感染与宫颈癌关系的分子流行病学研究中具有重要意义和广阔的应用前景。     

Human papillomavirus infection and neoplasia of the cervix and anogenital region in women with Hodgkin's disease

Immunosuppression has been associated with an increased incidence of condyloma and neoplasia of the cervix and anogenital region, lesions associated with human papillomavirus (HPV) infection. The role of immunosuppression as a risk factor for the development of these lesions in patients with Hodgkin's disease was studied in a series of 666 consecutive women seen at the University of Texas M. D. Anderson Hospital between 1963 and 1982. Thirty-nine patients (5.9%) of the entire series and 45.9% of the 85 women with Hodgkin's disease who had a gynecologic examination showed evidence of condyloma, dysplasia or carcinoma of the cervix or anogenital region. This is a significantly greater percentage of affected women than would be expected based on published results from Papanicolaou screening services. The evidence is compatible with immunosuppression from Hodgkin's disease or its treatment rendering a woman susceptible to persistent HPV infection and its consequences.     

Double cervix with bilateral and synchronous HSIL associated with different high-risk HPV types. A case report

BACKGROUND: High grade squamous intraepithelial lesion (HSIL) of the cervix is well known to be associated with human papillomavirus (HPV) infection. HSIL and invasive carcinomas occurring synchronously in genital malformations, such as a double cervix, have been reported. It has been postulated that the field effect phenomenon of HPV infection is responsible for this synchronous infection. However, there is no information in the literature on the specific types of HPV causing the concomitant lesions in cases with a double cervix. CASE: A 33-year-old nulligravida with a double cervix and a single uterine corpus was diagnosed with bilateral HSIL on Papanicolaou-stained ThinPrep slides (Cytyc Corp., Boxborough, Massachusetts, U.S.A.). A bilateral loop electrosurgical excision procedure cone biopsy revealed HSIL involving both cervices. DNA extracted from the HSIL lesions was analyzed by a polymerase chain reaction-based assay for the presence of HPV. High-risk HPV type 33 was identified in the right cervix, while HPV type 35 was present in the left. CONCLUSION: Demonstration of high-risk HPV types bilaterally supported the etiologic role of HPV infection in the synchronous and bilateral occurrence of HSIL in this case of a double cervix. The HPV types were different in the right and left cervices.     

Human papilloma virus associated with genital infection

Genital human papillomavirus (HPV) infections are among the most common sexually transmitted diseases. HPV is associated with a spectrum of diseases ranging from benign vulgar verrucae and condylomata accuminata to malignant cancers of the cervix, vulva, anus and penis. Genital HPV is in most cases transmitted sexually, but non-sexual routes of transmission, such as perinatal and autoinoculation, are possible. Men can be a reservoir of the virus that lives in latent or subclinical form on genital mucosa. Such an asymptomatic infection may be an oncogenic factor in the development of cervical cancer Colposcopic examination of the genitalia after the application of 3-5% acetic acid is a reliable method for the identification of subclinical HPV infection. Successful therapy of anogenital warts is characterized by their complete clearance, as well as by the lack of recurrence. Current treatments do not reliably eradicate HPV infections. The diagnosis and therapy of HPV infection in men is potentially beneficial because the eradication of penile HPV infection may decrease the reservoir of the virus.     

Comparison of the prevalence and distribution of human papillomavirus infection and cervical lesions between urban and native habitants of an Amazonian region of Peru

We made a study of the prevalence of human papillomavirus (HPV) and cervix lesions in an Amazonian Bora native population (Bn) and compared it with the prevalence in an urban population in Iquitos (Iq). We also examined the distribution of HPV types among abnormal cervical smears in the Iq population. Swabs and cytologies were collected from 472 females. DNA consensus PCR, followed by direct sequencing, were used to determinate the HPV types in the swabs. Cytologies were classified based on the lesion grade. HPV prevalence was 43.9% in Iq and 35.4% in Bn. Cervix lesion prevalence was 20.0% in Iq and 0.3% in Bn. The frequency of high-risk HPV types among HPV+ females was 71.9% in Iq and 56.3% in Bn. The frequencies of low-risk and undetermined risk HPV types were 19.3/6.3% (Iq/ Bn) and 12.3/37.5% (Iq/Bn), respectively. In lesIq (women seeking cytological services with a previous cervix lesion diagnosis) the prevalence of HPV, was 72.9%. The incidence of carcinoma and high-grade squamous intraepithelial cervix lesions in lesIq were 31.2 and 18.8%, respectively. The Bn population had a lower incidence of high-risk HPV and cervix lesions. The high-risk strain HPV16 was significantly more frequent in Iq compared with Bn. lesIq high-risk HPVs were more frequent in high-grade squamous intraepithelial lesions and carcinoma. High-risk HPV16 prevalence was significantly higher than the prevalence of the other high-risk HPVs, especially in the high-grade squamous intraepithelial lesions and carcinoma.     

Identification of human papilloma viruses in male dysplastic genital lesions

The association of Human Papillomaviruses (HPV) DNA with female genital lesions has been widely documented whereas little has been reported about male genital pathologies. The aim of this work was to investigate the presence of HPV DNA and the genotype involved in male dysplastic genital lesions. All samples were analysed by polymerase chain reaction (PCR) to amplify HPV E1 and L1 genes. The PCR products were subjected to restriction fragment length polymorphism (RFLP) to determine the HPV genotype. We analysed 209 male genital biopsies from different lesions: mostly from acuminate condylomata and from Buschke-Lowenstein tumours, Bowen papulosis, leukoplakia of the glans, scrotal lymphangioma, penile horn and penile/perianal verrucous carcinoma. Our results revealed the constant presence of viral DNA in genital condylomata, mainly associated with low risk HPV; the presence of the same genotypes was also detected in some of the examined rare pathologies.     

AgNOR technique could be useful for differential diagnosis of squamous carcinomatous cells and atypical cells related to human papillomavirus in cervical smears

The human papillomavirus (HPV)-associated effect can mimic invasive squamous cell carcinoma. Measurements of the argyrophilic nucleolar organizer region (AgNOR) area of cells from smears with HPV infection of the cervix with marked atypia were carried out to differentiate this pathology from keratinizing carcinomas. After destaining, the smears were incubated in the dark for 25 min with a mixture of silver nitrate and gelatin in formic acid. After washing with deionized water and sodium thiosulfate, the slides were dehydrated and mounted with Canada balsam. The average AgNOR area was determined by image cytometry using the immersion oil objective and selecting 100 cells in each smear. Twenty-three patients with a mean AgNOR area of 1.32 µm² among their samples showed normal colposcopies and cervical smears after 18 months. In four patients, whose samples were diagnosed as atypical squamous cells of undetermined significance with viral atypia, the average AgNOR area was 7.70 µm²; biopsies showed keratinizing squamous carcinomas in three of these cases and moderately differentiated squamous carcinoma in one of them. We propose a cutoff of 2.2 µm² for the AgNOR area of cells from smears with HPV infection of the cervix with marked atypia to differentiate this group from keratinizing carcinomas.     

空泡细胞在诊断宫颈人乳头瘤病毒感染中的意义

本文对空泡细胞(Koilocyte)的形态进行了光镜和电镜观察,并应用分子杂交技术和PAP染色的方法对宫颈疾患中人乳头瘤病毒(HPV)DNA和抗原进行了检测,分析HPV DNA及抗原检测结果与空泡细胞出现的关系发现,空泡细胞阳性的病例中HPV DNA及抗原的检出率较低,并且部分空泡细胞阳性的病例中也可测出疱疹病毒Ⅱ型(HSV_2)抗原,结果提示:空泡细胞并不是HPV感染所特有的,空泡细胞作为诊断宫颈HPV感染的特征性指标值得怀疑。     

Human papillomavirus DNA detection in Papanicolaou-stained cervical smears with a nonradioactive, in situ hybridization assay.

Archived Papanicolaou-stained cervical smears from women with different cervical pathologies were processed for human papillomavirus (HPV) DNA detection and typing with an in situ hybridization (ISH) assay that employed commercial biotinylated HPV DNA probes. Two HPV DNA probes were utilized: one included HPV genotypes 6/11 and the other, 16/18. The method yielded positive results for HPV DNA 6/11 in 5 cases with condylomata acuminata (100%) and in 2 of 47 with flat warty lesions (4.2%), whereas HPV DNA 16/18 was detected in 29/47 of the latter group (61.7%). In cases with cervical intraepithelial III or invasive squamous cell carcinoma the yield was lower: positive results for HPV DNA 16/18 were obtained in only one of the five cases with one or the other cervical pathology (20%). An analysis of the results showed that the sensitivity of the assay correlated with evidence in the Papanicolaou specimens of pathognomonic cell injury from HPV infection. In the presence of such cytologic features, HPV DNA typing was possible in 37/52 cases (65.4%). In view of the modest difficulty and relatively quick execution of the nonradioactive ISH assay, the authors believe that Papanicolaou cervical smears with cytologic changes of HPV infection could be processed by this method in order to acquire information on the HPV type or types involved in the cervical infection.     

1235例妇科门诊患者高危型HPV感染状况及亚型分布调查研究

目的了解本地区妇科门诊患者宫颈高危型HPV感染状况及亚型分布,为今后的宫颈癌前病变、宫颈癌防治提供临床依据。方法采用基因芯片技术对1 235例妇科门诊患者进行HPV筛查,筛查出的阳性患者应用流式荧光杂交法进行高危型HPV亚型检测,分析比较宫颈炎、宫颈鳞癌及宫颈腺癌患者高危型HPV感染情况及亚型分布差异。结果六安市金安区妇幼保健院妇科门诊患者HPV感染率高达60%,其中高危型HPV感染率为43. 2%,主要以HPV16、HPV18为主;低危型HPV感染率为30.0%,主要以HPV11为主;单一感染阳性率为34. 1%,而混合型感染高达65. 9%,且两者均以HPV16型和HPV18型为主。宫颈炎患者HPV16型、HPV18型及HPV16 ＋ HPV18型的检出率明显低于宫颈鳞癌和宫颈腺癌患者,其中宫颈腺癌患者HPV16 ＋ HPV18型混合感染率最高。结论妇科门诊患者HPV感染率较高,宫颈癌患者HPV16及18型检出率显著高于宫颈炎患者,加强HPV高危基因型的监测有助于预警宫颈癌尤其是宫颈腺癌的发生。     

Human papillomaviruses and DNA ploidy in anal condylomata acuminata

Previous studies have emphasized the usefulness of DNA ploidy measurement and Human Papillomavirus (HPV) detection as prognostic markers in low grade cervical lesions. We addressed the eventual relationship between HPV type, DNA profile, and p53 tumor suppressor protein expression in anal condylomata acuminata to eventually determine parameters which may be considered as predictive risk factors for the development of cancer. DNA ploidy was assessed by image cytometry after Feulgen staining of contiguous serial sections of 45 anal condylomata acuminata without atypia containing HPV detected by in situ hybridization and Polymerase Chain Reaction (PCR). p53 expression was detected by immunohistochemistry. DNA aneuploidy was found in 53.3% of these lesions, 48.9% containing non oncogenic HPV types 6 and/or 11 and 4.4% harbouring HPV types 11 and 18. The DNA diploid lesions were all associated with non oncogenic HPV types 6 and/or 11 and one case also contained HPV type 33. There was no significant correlation between the detection of DNA aneuploidy and the presence of immuno-detected p53. DNA aneuploidy was not related to the presence of oncogenic HPV in anal condylomata acuminata. The DNA aneuploid profile frequently observed, especially in lesions associated with non oncogenic HPV types, is not yet well explained and cannot be considered as a prognostic factor. In contrast, a more intensive clinical follow-up should be proposed in patients with oncogenic HPV associated to DNA aneuploidy.     

Detection of human papillomavirus (HPV) in genital warts and carcinomas by DNA in situ hybridization in Chinese patients

A series of 51 genital biopsies from normal epithelium, condylomata acuminata, leucoplakia and squamous cell carcinoma from Chinese male and female patients were analysed for the presence of human papillomavirus (HPV) types 6, 11, 16 and 18 by DNA in situ hybridization. All of the nine genital condyloma acuminata were positive for HPV DNA, in which HPV 6 was found in six cases, HPV 11 in two cases and HPV 18 in one case. Twelve out of the 21 cases (57.1% of the total) of cervical squamous cell carcinoma were shown to contain HPV DNA; HPV 16 was found in nine cases, HPV 18 in two cases and HPV 16/18 in one case. Present results support the earlier concept that HPV 6/11 are closely associated with benign genital lesions, and HPV 16/18 are mostly confined to higher grade of intra-epithelial neoplasias and carcinoma.     

实时荧光定量PCR检测HPV—DNA在宫颈癌筛查中应用

目的：探讨HPV感染与宫颈病变的关系。方法：对2008年2月-2009年3月期间在通山县人民医院皮肤性病科和妇科门诊就诊的1256位女性的宫颈拭子标本进行HPV DNA实时荧光定量PCR检测,比较不同宫颈病变级别组HPV的阳性率。结果：各病变组与正常组比较差异有显著性,P〈0.01,且随病变级别增加总阳性率逐渐上升。结论：HPV在人群具有较高的感染率,且HPV感染与宫颈病变的发生有关。实时荧光定量PCR检测HPV DNA可成为一种广泛应用的临床检验技术,作为筛查宫颈癌及癌前病变的首选方法。     

Value of cytology in papillary condylomatous lesions of the cervix

OBJECTIVE: To determine the value of cytology in detecting mature and immature papillary condylomas of the uterine cervix. STUDY DESIGN: We evaluated 240 cases of plane cervical intraepithelial neoplasia, grade 1 (CIN 1), and 23 papillary condylomas by Pap smear and biopsy and classified histologic sections according to maturity and keratinization. We reevaluated corresponding cytologic smears and identified criteria of low grade squamous lesion (LSIL) and human papillomavirus (HPV) infection. RESULTS: Thirteen (56.5%) papillary lesions were histologically classified as mature, 6 (26%) as immature and 4 (17.3%) as mixed. Fifteen lesions (65.2%) were nonkeratinized, 2 (8.6%) keratinized and 6 (26%) partially keratinized. Corresponding smears were cytologically diagnosed as LSIL (6, 26%), atypical squamous cells of undetermined significance (ASCUS) (7, 30.4%) and negative (10, 43.4%). Careful cytologic review diagnosed only two of the 13 mature lesions; few cytological criteria of LSIL and HPV infection were observed. Koilocytes were seen in just 1 case. Sample limiting factors occurred in 4 cases: 2 cytologically diagnosed as LSIL, 1 asASCUS and 1 as negative for lesion. CONCLUSION: Cytology was not effective in the detection of cervical condyloma acuminatum, independent of limitations in sample adequacy and of the degree of maturity or keratinization of the lesions.     

Cervical squamous intraepithelial lesions and associated cervical infections in an HIV‐positive population in Rural Mpumalanga,South Africa

P. J. Swanepoel, P. Michelow, R. Du Plessis, I. G. Proudfoot, G. A. Tarr, S. L. Bockel, C. J. Swanepoel
Cervical squamous intraepithelial lesions and associated cervical infections in an HIV‐positive population in Rural Mpumalanga, South Africa Background: The incidences of genital human papillomavirus (HPV) infection, associated squamous intraepithelial lesions and cervical squamous cell carcinoma are significantly increased in HIV‐positive women. The role of other cervicovaginal infections in the acquisition of the HPV infection, cervical carcinogenesis and genital HIV infection remains largely speculative. Methods: A retrospective study was conducted including 1087 HIV‐positive women in rural Mpumalanga province, South Africa, for the period 1 May 2009 to 31 August 2010. For each patient, the age at first presentation, cervical cytological diagnosis, subsequent follow‐up cytology and histology, and microscopically visible infections (including endemic Bilharzia) were tabulated and statistically analysed. Results: The prevalence of low‐grade squamous intraepithelial lesion (LSIL), high‐grade squamous intraepithelial lesion (HSIL), squamous cell carcinoma, atypical squamous cells of undetermined significance (ASC‐US) and atypical squamous cells, cannot exclude HSIL (ASC‐H) in the study population were 22.1%, 30.9%, 0.6%, 13.5% and 4.0%, respectively. LSIL, HSIL and squamous cell carcinoma were diagnosed, respectively, at the average ages of 35.7, 37.9 and 37.2 years. Four patients with cervical intraepithelial neoplasia grade 1 (CIN1), 32 with CIN2/CIN3 and two with cervical squamous cell carcinoma were also diagnosed with Bilharzia. Of the other infections only bacterial vaginosis had a positive statistical correlation with HPV‐induced cervical abnormalities (LSIL, HSIL or squamous cell carcinoma). Conclusion: This study confirms the high prevalence of progressive HPV‐associated cervical disease in a rural Southern African HIV‐positive population, which is at least equal to or worse than in other African HIV‐positive studies. The high incidence of Bilharzia infection in those cases that underwent cervical cone excision suggests a possible relationship with progressive HPV disease and cervical carcinogenesis. Bacterial vaginosis (perhaps in combination with Bilharzia) may compromise the normal barriers against HPV and HIV infection.     

Human Papillomavirus Genotype Distribution in Czech Women and Men with Diseases Etiologically Linked to HPV

Background

The HPV prevalence and genotype distribution are important for the estimation of the impact of HPV-based cervical cancer screening and HPV vaccination on the incidence of diseases etiologically linked to HPVs. The HPV genotype distribution varies across different geographical regions. Therefore, we investigated the type-specific HPV prevalence in Czech women and men with anogenital diseases.

Methods

We analyzed 157 squamous cell carcinoma samples, 695 precancerous lesion samples and 64 cervical, vulvar and anal condylomata acuminate samples. HPV detection and typing were performed by PCR with GP5+/6+ primers, reverse line blot assay and sequencing.

Results

Thirty different HPV genotypes were detected in our study, HPV 16 being the most prevalent type both in precancerous lesions (45%) and squamous cell carcinomas (59%). In benign lesions, HPV 6 (72%) was the most common type. Altogether, 61% of carcinoma samples and 43% of precancerous lesion samples contained HPV 16 and/or 18. The presence of HPV types related to the vaccinal ones (HPV 31, 45, 33, 52, 58) were detected in 16% of carcinoma samples and 18% of precancerous lesion samples. HPV 16 and/or 18 were present in 76% of cervical cancer samples, 33% of CIN1, 43% CIN2 and 71% of CIN3 samples. HPV types 6 and/or 11 were detected in 84% samples of condylomata acuminate samples.

Conclusions

The prevalence of vaccinal and related HPV types in patients with HPV-associated diseases in the Czech Republic is very high. We may assume that the implementation of routine vaccination against HPV would greatly reduce the burden of HPV-associated diseases in the Czech Republic.     

Glandular and squamous atypia and intraepithelial lesions in atrophic cervicovaginal smears. One institution's experience

OBJECTIVE: To review the cytologic features and follow-up histologic findings in atrophic cervicovaginal smears with the diagnoses of glandular or squamous atypia or intraepithelial lesion. STUDY DESIGN: A total of 228 cases were included in the study. The selection criteria included: age > 48 years and a diagnosis of either atypical glandular cells (AGC) (51 cases), cellular changes suggestive of human papillomavirus (HPV) infection (S/O HPV, 97 cases), low grade squamous intraepithelial lesion (LSIL) (60 cases) or high grade squamous intraepithelial lesion (HSIL) (20 cases). Follow-up biopsy information was available for 103 cases (45%). RESULTS: From the AGC group, 35 (69%) cases had tissue studies; 14 (40%) cases showed glandular lesions; 5 (14%) showed squamous intraepithelial lesion (SIL) and atypical cells. Follow-up information was available for 32 (33%) cases classified as S/O HPV; significant lesions (glandular/squamous) were found in 11 (34%). In the LSIL category, 22 (37%) cases had follow-up; 16 (73%) showed SIL. In the HSIL category, 14 cases (70%) underwent biopsy, and all showed SIL (four LSIL and nine HSIL) or squamous cell carcinoma. CONCLUSION: Even though atrophy-related epithelial changes often pose diagnostic difficulties in the interpretation of postmenopausal smears, application of reproducible and established cytologic criteria in diagnosing SIL and/or glandular lesions can improve diagnostic accuracy and result in selection of patients for follow-up tissue studies.     

Human papillomavirus infection and its role in the genesis of dysplastic and neoplastic lesions of the squamous epithelia

Extensive laboratory and epidemiological evidence demonstrate that human papillomavirus (HPV) is the major cause of squamous cervical carcinoma (SCC), its precursor lesions (cervical intraepithelial neoplasia - CIN) and several other benign and malign clinical manifestations including genital warts, condylomata acuminata, Bowenoid papulosis, vaginal, vulvar and anal intraepithelial neoplasia (VIN and AIN) and carcinoma, penile carcinoma and other squamous neoplasias of the head and neck districts. In addition, mother-to-child transmission is probably responsible for recurrent laryngeal and pulmonary papillomatosis in infants. The relevance and high level of scientific interest surrounding HPVs are related to the oncogenic potential of some viral types belonging to this family and the possibility to influence the incidence of various tumour forms likecervical carcinoma, improving the efficacy of specific screening programs or defining preventive strategies like vaccination.     

人乳头瘤病毒与宫颈癌前病变的相关性研究

目的：探讨人乳头瘤病毒感染与宫颈癌前病变两者之间的相关性。方法：选择2009年11月～2011年12月来我院就诊的220例宫颈病变患者为研究对象,将患者分成4组,即炎症组、CINl组、CIN2组和CIN3组,各组进行人乳头瘤病毒检出率及病毒载量的比较。结果：炎症组、CINl、CIN2和CIN34组人乳头瘤病毒的检出率分别为67．2％、80％、87．3％和91．4％且4组之间比较差异有显著统计学意义（P〈O．001）。炎症组HPV感染的阳性率与CIN组比较显著降低（P〈0．005）,宫颈癌前病变各组HPV感染呈阳性的病毒载量差异有统计学意义（P〈0．05）。结论：宫颈癌前病变的严重程度和感染高危型HPV病毒载量呈正相关,随着感染高危型HPV病毒载量越高,宫颈癌前病变的程度越严重。