应用计算机辅助细胞检测系统检测子宫颈涂片中人乳头状瘤病毒感染

目的评价计算机辅助细胞检测（ｃｏｍｐｕｔｅｒａｓｉｓｔｅｄｃｙｔｏｌｏｇｉｃｔｅｓｔ,ＣＣＴ）系统检测宫颈涂片中人乳头状瘤病毒（ｈｕｍａｎｐａｐｉｌｏｍａｖｉｒｕｓ,ＨＰＶ）感染的敏感性与准确性。方法对１５８例外阴尖锐湿疣患者的子宫颈涂片,采用ＣＣＴ检测技术进行细胞病理学诊断,其诊断标准依据ＴｈｅＢｅｔｈｅｓｄａＳｙｓｔｅｍ（ＴＢＳ）分类法。同期行阴道镜指引下的子宫颈活组织检查和聚合酶链反应（ＰＣＲ）技术检测宫颈拭子中的ＨＰＶＤＮＡ。结果ＣＣＴ诊断宫颈ＨＰＶ感染的敏感性：与组织学检查对照,为７４６２％;与ＰＣＲ检测对照,为６９１１％。准确性：与组织学检查对照,为６７７２％;与ＰＣＲ检测对照,为７０８８％。结论ＣＣＴ作为一种细胞病理学检测技术,用于诊断宫颈ＨＰＶ感染或亚临床感染,有临床应用价值     

Detection of human papillomavirus DNA in cervical smears by the polymerase chain reaction

Abstract. Chetsanga C, Pettersson B, Pettersson U, Gyllensten U. Detection of human papillomavirus DNA in cervical smears by the polymerase chain reaction. Int J Gynecol Cancer 1991; 1 : 209–213.
A PCR-based method has been developed which allows human papillomavirus DNA to be detected in routinely obtained cervical smear specimens. The results showed that 76% of the smears that were collected from women who were carrying HPV-positive lesions by cervical swab analysis were positive by the PCR-based method. The results suggest that it is possible to carry out population-based retrospective studies of archival cervical smears.     

Detection of human papillomavirus DNA in cervical lesions by in situ hybridization using biotinylated DNA probes

In situ hybridization (ISH) for human papillomavirus (HPV)-6, -11, -16, -18, and -31 DNA was performed on 615 formalin-fixed paraffin-embedded cervical biopsies using biotinylated DNA probes. Results were obtained from 584 samples with 266 (45.5%) containing HPV-DNA sequences. Ninety percent of condyloma acuminatum specimens were positive for HPV-DNA with 18 of 19 positive cases containing HPV-6 or -11 DNA. The detection rate of HPV in cervical intraepithelial neoplasia (CIN) lesions was 50.6% (239 of 472), while only 8 of 91 (8.9%) cervical biopsies considered to be histologically normal or with minimal dysplasia contained HPV-DNA as demonstrated by ISH. The prevalence of HPV-16, -18, and/or -31 DNA increased with the severity of the lesions, with 20 of 20 (100%) positive CIN-III lesions containing these viral types compared with 102 of 157 (65.0%) positive CIN-I lesions. ISH with biotinylated DNA probes appears helpful in identifying lesions containing higher risk viral strains.     

Detection of human papillomavirus DNA in human cervical lesions by tissue in situ nucleic acid hybridization

Cervical cancer is one of the most common female cancers in Taiwan. Certain types of human papillomavirus (HPV) are frequently detected in the epithelial precancerous and cancerous lesions of the cervix. By the use of tissue in situ hybridization, we investigated the relationship of various types of HPV (group I, HPV-6 & 11, group II, HPV-16 & 18, group III, HPV-31, 33 & 35) with cervical condyloma, carcinoma as well as precancerous lesions. Group I HPV DNAs were mainly found in cervical condylomatous lesions (2/2) of the cervix and cervical intraepithelial neoplasia I (CIN I) (2/4), but were only occasionally found in CIN II (1/4), CIN III (1/9) or non-keratinized squamous cell carcinoma (1/15). HPV DNAs of groups II and III were mainly detected in lesions of CIN III (5/9) and invasive squamous cell carcinoma (large cell, keratinized type: 4/7; large cell, non-keratinized type: 11/15). HPV DNA sequences were invariably detectable only in the cell nuclei of condyloma or dysplastic epithelium or invasive carcinoma. However, they could not only be detected in the upper layer dysplastic cells and koilocytes but also in the well and poorly differentiated cervical cancer cells. The distribution of HPV DNA positive cells in the carcinomas fell into four different patterns: (1) upper zone and non-invasive regions of the carcinoma (11/22, 50%), (2) basal zone and invasive regions (2/22, 9%), (3) randomly scattered (7/22, 32%), and (4) extensively distributed over the whole tumor lesions (2/22, 9%). Thus, our results are consistent with a strong correlation between the presence of HPV-16, 18, 31, 33 and 35 and malignant conversion of cervical epithelial cells.     

高危型人乳头状瘤病毒DNA检测与细胞学联合检查对子宫颈癌前病变筛查的研究

目的评价高危型人乳头状瘤病毒(HPV)DNA检测联合宫颈细胞学检查对宫颈癌前病变筛查的效果。方法2003年1月至2004年6月,对广东省人民医院妇科门诊5210名妇女进行宫颈癌前病变筛查,采用第二代杂交捕获试验(HC-Ⅱ)检测高危型HPV DNA联合细胞学检查,同时进行阴道镜检查,并以宫颈活检的组织病理学结果为确诊标准。结果受检者平均年龄(34±9)岁,筛查并最后经病理诊断为HPV感染890例,宫颈上皮内瘤变(CIN)Ⅰ级83例,CINⅡ73例,CINⅢ80例,宫颈浸润癌54例,子宫内膜癌5例,阴道上皮内瘤样病变1例,宫颈结核1例。以组织病理学为确诊标准,高危型HPV DNA检测CINⅡ、Ⅲ的敏感度是92·22%,特异度是74·71%,阳性预测值5·19%,阴性预测值99·84%。宫颈细胞学筛查CINⅡ、Ⅲ,以未明确诊断意义的不典型鳞状上皮细胞(ASCUS)为分界点的敏感度、特异度、阳性预测值和阴性预测值分别是90·00%、80·34%、11·94%和99·63%;以低度鳞状上皮内病变(LSIL)为分界点的敏感度、特异度、阳性预测值和阴性预测值分别是70·13%、91·58%、11·11%和99·51%;以高度鳞状上皮内病变(HSIL)为分界点的敏感度、特异度、阳性预测值和阴性预测值分别是48·05%、98·46%、31·90%和99·21%。高危型HPV DNA检测联合细胞学检查筛查CINⅡ、Ⅲ的敏感度、特异度、阳性预测值和阴性预测值分别是98·70%、73·08%、5·21%和100·00%。高危型HPV DNA在不同宫颈病变中的阳性率分别是:宫颈癌85·2%(46/54),CINⅢ92·5%(74/80),CINⅡ86·3%(63/73)和CINⅠ45·8%(38/83)。结论高危型HPV DNA检测在宫颈癌前病变的筛查中有很高的敏感度和阴性预测值,高危型HPV DNA检测联合细胞学检查可使敏感度和阴性预测值有提高,但特异度未能提高。     

Influence of ovarian stimulation on the detection of human papillomavirus DNA in cervical scrapes obtained from patients undergoing assisted reproductive techniques

OBJECTIVE: To determine whether gonadotropin stimulation influences the detection of human papillomavirus (HPV) DNA in cervical scrapes. DESIGN: Prospective, controlled study. SETTING: Tertiary care infertility clinic. PATIENT(S): Two hundred ninety-four patients enrolled in an IVF or IUI program. Two thousand two hundred sixty-two women from an ongoing screening study who were of similar age served as a control group. INTERVENTION(S): Cervical scrapes were obtained with a cytobrush before and after ovarian stimulation with gonadotropins. MAIN OUTCOME MEASURE(S): Human papillomavirus status was assessed with a general primer (GP) polymerase chain reaction (PCR) using the GP5+/GP6+ system. In GP-PCR-positive samples, high-risk HPV types were identified with a cocktail of digoxigenin-labeled oligonucleotides. Viral load was evaluated by semiquantitative analysis of the PCR products. RESULT(S): The prevalence of high-risk HPVs was 7.8% before stimulation and 6.8% after stimulation and, thus, was similar to the prevalence in controls (8.4%). Twenty-nine patients were positive for high-risk HPVs: 14 were positive before and after stimulation, 6 were negative before and positive after stimulation, and 9 were positive before and negative after stimulation. Positivity for high-risk HPVs and viral load did not correlate directly with serum estrogen levels. CONCLUSION(S): Ovarian stimulation has no significant effect on the prevalence of HPV DNA in cervical scrapes obtained from patients undergoing assisted reproductive techniques.     

Detection of human papillomavirus DNA in biopsy-proven cervical squamous intraepithelial lesions in pregnant women.

This study analyzed cervical squamous intraepithelial lesions (SILs) in pregnant women for human papillomavirus (HPV) using in situ hybridization analysis. HPV DNA was detected in 77% (23/30) of low-grade SILs as compared to 89% of such lesions in nonpregnant women. The detection rate in high-grade SILs was 41% (9/22) compared to 70% in nonpregnant women. Analysis by the polymerase chain reaction showed that the detection rates were similar (96-100%) for low- and high-grade lesions in pregnant and nonpregnant women, which demonstrates that in situ negative tissues indeed contained HPV DNA. Low-grade SILs contained a heterogeneous group of at least 14 different HPV types, whereas most high-grade SILs contained HPV 16. We concluded that cervical SILs in pregnant women are invariably associated with HPV. Further study is needed to determine which of several possible variables, such as the age of the lesion and the viral copy number, may explain the apparent decreased detection rate of HPV by in situ hybridization in SILs during pregnancy.     

Cytologic correlates of cervical papillomavirus infection

A fundamental question in Papanicolaou smear screening is the specificity of cytologic criteria for the recognition of genital human papillomavirus (HPV) infection. To address this problem, we conducted a two-phase study of routinely screened women to determine the efficiency with which cytologic findings identified the presence of HPV DNA, focusing on the criteria for identifying smears as "atypical." In phase 1, 25 of 290 (8.6%) smears were designated atypical, but only 3 (12%) of the samples contained HPV nucleic acids. Four of five (80%) smears designated as diagnostic of HPV/cervical HPV infection were associated with HPV nucleic acids. By applying more stringent criteria for the diagnosis of atypical in phase 2, only 3 of 166 (1.8%) were identified as atypical. Of these, two (67%) contained HPV nucleic acids. The criteria that most efficiently correlated with HPV nucleic acids included prominent nuclear enlargement with either multiple nuclei or nuclear hyperchromatism. On review of the 19 HPV-positive and 20 control HPV-negative smears originally diagnosed as cytologically negative, the above criteria identified an additional 3 cytologically atypical/positive smears versus none (0 of 20) in the control group. This study supports the concept that cytologic abnormalities suggesting "subtle" HPV infection may be extremely difficult to distinguish from non-HPV-related changes, and that criteria used to imply "suggestive but not diagnostic for HPV infection" should be continually reevaluated. The potential role of HPV DNA analysis in Papanicolaou smear interpretation is discussed.     

宫颈疾病HPV混合型感染的检测与分析

目的:探讨人乳头状瘤病毒(HPV)混合型感染导流杂交法检测宫颈疾病的价值。方法:用导流杂交法,检测本地区304例妇女(其中宫颈癌组35例、宫颈上皮内瘤变组20例、炎症组140例和正常组109例)宫颈脱落细胞21种HPV基因亚型,分析各组患者HPV多重感染的特点。结果:304例标本中检出HPV阳性94例,包括混合型30例,HPV多重感染检出率在宫颈癌组、宫颈上皮内瘤变组、炎症组和正常组分别为34.3%、20.0%、7.1%和2.8%,各组HPV混合型感染率差异有统计学意义(χ2=463.579,P<0.001);混合型感染中检出的HPV亚型由高到低依次为:HPV-16(19.2%)、HPV-52(13.7%)、HPV-58(12.3%)、HPV-18(8.2%)、HPV-11(6.8%)、HPV-6(6.8%)、HPV-31(5.5%)、HPV-33(5.5%)、HPV-66(5.5%)、HPV-39(4.1%)、HPV-53(4.1%)、HPV-68(4.1%)、HPV-56(1.4%)、HPV-59(1.4%)、CP8304(1.4%)。各疾病组检出的混合型HPV感染前3位HPV基因型均为高危型。结论:宫颈癌HPV多重感染率高,主要系高危HPV型感染所致;导流杂交法HPV混合型感染检测值得在宫颈疾病诊断中推广应用。     

Clinical significance of serum human papillomavirus DNA in cervical carcinoma

OBJECTIVE: To study the association between serum human papillomavirus (HPV) deoxyribonucleic acid (DNA) and clinicopathologic prognostic factors and the clinical usefulness of serum HPV DNA in early-stage cervical cancer. METHODS: Deoxyribonucleic acids extracted from cervical tissues and sera of patients with stage IB or IIA cervical cancer and 40 controls including patients with cervical carcinoma in situ or benign disease were examined for HPV DNA with L1 consensus and types 16- and 18-specific E7 primers. Multivariable logistic regression was used to determine significant correlates of positive serum HPV DNA, and the receiver operating characteristic curve was applied in risk-factor assessment. RESULTS: Human papillomavirus DNA was not detected in sera from patients with carcinoma in situ or benign disease. Among the 112 patients with cervical cancer, we detected 27 positive samples (24.1%) in serum. Positive HPV DNA in serum was significantly associated with lymphovascular invasion and deep stromal invasion with or without parametrial extension (P <.001 for both conditions), pelvic lymph nodal metastasis (P =.001), large tumor size, and elevated levels of serum squamous cell carcinoma antigen (P <.001 for both conditions). When serum HPV DNA was used to predict high-risk patients who require adjuvant therapy, a sensitivity of 45.2%, a specificity of 88.6%, a positive predictive value of 70.4%, and a negative predictive value of 72.9% were obtained. CONCLUSION: The presence of serum HPV DNA in patients with early-stage cervical cancer was correlated with poor prognosis factors that warrant adjuvant therapy.     

Amplified DNA detection of HPV types 16 and 18 DNA in cervical scrapes by polymerase chain reaction

Cervical scrapes from 43 patients (12 with chronic cervicitis, 13 with dysplasia, 11 with carcinoma in situ and 7 with invasive carcinoma) were examined by the polymerase chain reaction (PCR), to detect the amplified E7 gene of the HPV types 16 and 18 DNA sequence. HPV types 16 and/or 18 DNA were detected in 25 of 43 cases by the PCR (type 16 in 16, type 18 in 8, and both types 16 and 18 in 1). In comparison with the results for histological grades, the positive rate significantly increased as the grade of cervical dysplasia became higher. All cases were also examined with a HPV detection kit, the "Vira Pap" which contains 32P-labeled mixed RNA probes complimentary to HPV types 6, 11, 16, 18, 31, 33, 35 DNA. Six of 18 cases in which HPV DNA were not detected by the Vira Pap were positive for HPV types 16 and/or 18, so that the PCR was a highly sensitive method compared to the Vira Pap. The PCR has some advantages: 1. it requires only a small amount of specimens and 2. paraffin-embedded sections can also be used. We suggest that the PCR is a useful method for the screening and for retrospective investigation of HPV infection.     

宫颈癌及上皮内瘤变人乳头瘤病毒基因型的检测

目的:了解宫颈癌及上皮内瘤变人乳头瘤病毒(HPV)的感染率及其基因型的分布。方法:用PCR-RFLP法检测239例宫颈癌及上皮内瘤变患者HPV感染并进行分型。先用PGMY09/11共同引物扩增生殖道粘膜型HPV L1区的高度保守区,然后联合使用RsaⅠ、MseⅠ、PstⅠ和HaeⅢ4个限制性内切酶对阳性PCR产物进行酶切,利用不同的酶切片段鉴定HPV的基因型。结果:在239例宫颈癌及上皮内瘤变患者中共检出205例(85·8%)HPV感染,其中宫颈上皮内瘤变Ⅰ级(CINⅠ)、宫颈上皮内瘤变Ⅱ~Ⅲ级(CINⅡ~Ⅲ)和宫颈癌中HPV感染率分别是66·7%,89·9%和98·3%,差异有统计学意义(P<0·001)。在宫颈癌及上皮内瘤变中共检出22型HPV,其中主要基因型及其感染率分别是HPV16(45·6%)、58(12·1%)和52(6·3%)。结论:宫颈癌及上皮内瘤变中HPV感染的基因型至少可达22型,其中以HPV16、58和52为最常见。     

Detection of human papillomavirus DNA in advanced epithelial ovarian carcinoma

Tissue specimens from 10 out of 12 patients with advanced epithelial ovarian adenocarcinoma contained DNA of human papillomavirus type 6 (HPV-6). HPV DNA was identified by in situ hybridization at high stringency using biotin-labeled DNA probes. Nonneoplastic tissue specimens from other pelvic sites of the same patients were also examined. None showed evidence of HPV DNA. The meaning of these findings in relation to epithelial ovarian carcinoma is discussed.     

Cost-effectiveness of human papillomavirus DNA testing for cervical cancer screening in women aged 30 years or more

OBJECTIVE: To evaluate the cost-effectiveness of human papillomavirus (HPV) DNA testing as a primary screening test in combination with cervical cytology in women aged 30 years or more. METHODS: A state-transition mathematical model was used to simulate the natural history of HPV and cervical cancer in a cohort of U.S. women. Strategies included no screening and screening at different frequencies with conventional cytology, liquid-based cytology with HPV testing used for triage of equivocal results, and HPV DNA testing and cytology in combination after women had reached the age of 30. Outcomes measured included cancer incidence, life expectancy, lifetime costs, and incremental cost-effectiveness ratios. RESULTS: The estimated reduction in lifetime risk of cervical cancer varies from 81% to 93% depending on the screening frequency, type of cytology, and test strategy. Every 3-year screening with liquid-based cytology administered to women at all ages and every 3-year screening using HPV DNA testing and cytology in combination administered to women aged 30 years or more provide equivalent or greater benefits than those provided by annual conventional cytology and have incremental cost-effectiveness ratios of US dollars 95300 and US dollars 228700 per year of life gained, respectively. In comparison, annual screening with HPV DNA testing and cytology in combination provides only a few hours of additional life expectancy and has a cost-effectiveness ratio of more than Us dollars 2000000 per year of life gained. CONCLUSIONS: For women aged 30 years and more, every 2- or 3-year screening strategy that uses either HPV DNA testing in combination with cytology for primary screening or cytology with reflex HPV DNA testing for equivocal results will provide a greater reduction in cancer and be less costly than annual conventional cytology.     

Prevalence and risk factors for human papillomavirus DNA in cervical cytology

Objectives

This study aimed to detect the presence and prevalence of HPV-DNA in the cervical swab samples obtained from patients with cervical cancer, premalignant cervical lesions and benign cervical smear results, and to identify the potential risk factors influencing this prevalence.

Study design

Smear preparations were examined and classified according to the Bethesda system. HPV-DNA detection and genotyping was carried out using polymerase chain reaction combined with reverse hybridization line-probe assays. Age, smoking habit, age at first sexual intercourse, number of sexual partners, number of term births, contraceptive method, progesterone therapy, history of sexually transmitted diseases, history or existence of warts, existence of cervical infection and the history of circumcision of male sexual partners were recorded.

Results

Six hundred and forty-two women (96 women with abnormal cervical cytology and 546 women with normal cytology) provided cervical samples. Multiplex PCR testing revealed that prevalence of HPV-DNA was 38.9% in our study population. HPV-DNA was detected in 78.3% of the women with cervical cancer and 76.9% of the women with HGSIL. Abnormal cervical cytology was observed in 30% of HPV-DNA positive cases and in 5.4% of HPV-DNA negative cases. Our findings also indicate that smoking habit, number of sexual partners, history of sexually transmitted diseases, and abnormal cervical cytology were associated with HPV infection. With respect to parity, there was a decreased risk of HPV infection with the increase in the number of births.

Conclusions

Estimates of the prevalence of HPV infection vary greatly around the world, so the factors that contribute to the rare occurrence of cervical cancer after HPV infection might also differ from country to country. Information gathered from this study could be used to prioritize limited screening and treatment services given to woman who have specific characteristics that may put them at an increased risk of HPV disease.     

High-risk human papillomavirus DNA testing and high-grade cervical intraepithelial lesions

OBJECTIVE: To explore the role of high-risk human papillomavirus (HPV) DNA testing in the improvement of the recognition of cervical cancer and precancerous lesions in women with abnormal cervical cytology. METHODS: A total of 2152 women with abnormal cervical cytology were submitted to both HPV DNA testing and biopsy guided by colposcopy and the results were correlated. RESULTS: Positive rate of high-risk HPV DNA in groups of atypical squamous cells of undetermined significance (ASC-US), atypical squamous cells, cannot exclude high-grade (ASC-H), low-grade squamous intraepithelial lesions and high-grade squamous intraepithelial lesions was 53.7, 53.2, 84.6 and 93.0%, respectively. In each group, the detection rate of grade 2,3 cervical intraepithelial neoplasia (CIN 2,3) or cervical cancer in patients with positive HPV DNA was significantly higher than that with negative HPV DNA (P<0.05). In ASC-US group, the negative predictive value of high-risk HPV DNA testing for detection of CIN 2,3 and cervical cancer was 99.8% and the sensitivity 98%. CONCLUSION: HPV DNA testing is a useful indicator in the management of patients with ASC-US and plays an important role in the evaluation of risk for CIN 2,3 and cervical cancer.     

Prevalence of human papillomavirus DNA in cervical tissue. Retrospective analysis of 855 cervical biopsies

The histopathologic features of 855 cervical biopsies were correlated with the presence of human papillomavirus DNA using in situ hybridization (ISH) with biotin labeled type specific probes for Human Papilloma Virus (HPV) types 6, 11, 16, 18, 31, 33 and 51. HPV-DNA was found in 18% (13/72) of cervical intraeptihelial neoplasia I (CIN I), 30% (35/115) of CIN II, 28% (57(206) of CIN III, in 84% (21/25) of flat condyloma and in 13% (15/112) of normal cervical tissue. HPV DNA was detectable in 11% (5/46) of cervical adenocarcinoma and in 21% (59/279) of squamous cell carcinoma (SCC) of the cervix. High risk HPV types were identified more often than low risk HPV types in CIN I, CIN II, CIN III and SCC. HPV type 16/18 predominates over HPV type 31/33/51 in CIN I, flat condyloma and in SCC. The prevalence of HPV was strongly associated with the grade of differentiation of SCC. It was identified in 59% (23/39) of well differentiated SCC, in 18% (25/142) of moderately differentiated and in 11% (11/98) of poorly differentiated SCC. Received: 29 March 1996 / Accepted: 15. August 1996     

应用聚合酶链反应技术测定正常子宫颈组织人乳头瘤病毒DNA的研究

应用兼并复合人乳头瘤病毒一聚合酶链反应（ＨＰＶ－ＰＣＲ）技术对１４８例外观正常的子宫颈分泌物拭子标本进行人乳头瘤病毒（ＨＰＶ）ＤＮＡ的测定。结果：正常子宫颈ＨＰＶＤＮＡ检出率为３１７６％（４７／１４８）。其中ＨＰＶ－６，１１型为６．７６％（１０／１４８）；ＨＰＶ－１６，１８型为３．３８％（５／１４８）；ＨＰＶ其它型为２１．６２％（３２／１４８）。外观正常的子宫颈，ＨＰＶ感染的主要易感年龄为２２～３４岁的青年妇女。本研究对女性生殖道ＨＰＶ感染的传播途径进行了探讨。