液基细胞学技术联合高危HPV-DNA检测对宫颈癌前病变的诊断效度

目的: 探讨液基细胞学检查技术(LCT)联合高危人乳头状瘤病毒DNA(HPV-DNA)检测诊断宫颈癌前病变的效度。方法: 对2007年12月~2010年12月来我院行体检的19~65岁的6 521名女性采用LCT进行宫颈癌的筛查,以及HPV分型基因芯片检测系统进行18种高危HPV基因亚型检测。对上述检测阳性者行阴道镜下宫颈活组织检查,对检测均阴性者依其意愿进行阴道镜下宫颈活组织检查。结果: LCT阳性(≥ASCUS)152例,HPV阳性86例,其中二者均为阳性的有42例,LCT和HPV均为阴性的有6 325例;LCT阳性的152例和HPV阳性的86例中病理活组织检查结果为阳性(≥CIN I)的分别有112例和68例,其中LCT和HPV均阳性的42例中病理活组织检查阳性的有34例。LCT和HPV均为阴性的6 325例中有2 000人自愿行病理检查,其中1人病理检查结果为阳性。LCT诊断宫颈癌前病变的灵敏度为76.19%,特异度为98.05%;HPV检测诊断宫颈癌前病变的灵敏度为46.26%,特异度为99.12%;两方法联合诊断(其中1项阳性即判定为患者)宫颈癌前病变的灵敏度为99.32%,特异度为99.61%。结论: 液基细胞检测技术和高危HPV-DNA检测的联合应用优于单项技术检测,对于宫颈癌前病变的筛检具有重要意义。     

宫颈癌及癌前病变应用HPV-DNA亚型检测联合液基细胞学检测的准确性

目的测定宫颈癌（Cc）癌组织和非癌组织的HPVcccDNA含量,并联合联合液基细胞学进行检测,探讨癌组织HPVcccDNA与cc的关系。方法采用荧光定量PCR检测45例宫颈癌患者癌组织和非癌组织中总HPVDNA和HPVcccDNA含量,分析2种组织中HPVcccDNA和总HPVDNA与癌组织病理分级的关系。结果血清HBsAg阳性的癌组织较对应非癌组织总HPVDNA无显著性差异（4.33士1.1μS,4.13±1.09,P=0.439）,而癌组织HPVcccDNA水平显著升高（3.09士1.22μS 2.62士0.97,P=0.039）,HPVcccDNA所占比例也显著升高（70.09％μS 62.33％,P=0.015）;42例中有14例（33％）的癌组织HPVcccDNA几乎成为HPVDNA唯一存在形式;3例血清HBsAg阴性的cc中均检测到了HPVcccDNA的存在,且HPVcccDNA比例均较高;癌组织和非癌组织HPVcccDNA与组织总HPVDNA均呈正相关（r=0.8244,P〈0.001;r=0.8460,P〈0.001）：癌组织中HPVcccDNA定量和肿瘤的病理分级无明显相关性（P〉O．05）。结论宫颈癌组织中HPVcccDNA水平和所占比例较非癌组织均升高,HPVcccDNA在宫颈癌变中有重要的作用,但水平与宫颈癌的病理分级无明显相关。     

9012例子宫颈液基细胞学诊断结果分析

目的探讨液基薄层细胞(TCT)检查在宫颈癌及癌前病变筛查中的应用价值。方法回顾分析开县人民医院9 012例宫颈液基细胞学检测结果和伯塞斯达系统(TBS)分类系统结果,其中12例ASC-US、10例ASC-H、35例低度鳞状上皮内病变(LSIL)、72例高度鳞状上皮内病变(HSIL)、2例鳞状上皮细胞癌(SCC)和6例腺细胞异常做阴道镜下病理活检,对细胞学异常结果与阴道镜下活组织检查病理诊断结果进行比较。结果 9 012例液基细胞学检测中,筛查出非典型鳞状细胞(ASC-US)以上病例285例,阳性率3.2%。液基细胞学检测与阴道镜下病理活检结果的符合率为91.24%。结论 TCT制片技术和TBS报告能较全面反映宫颈病变的情况,通过定期正规的筛查,能早期发现宫颈癌及癌前病变,从而早期治疗,阻止病变升级是预防宫颈癌的关键。     

15 393例宫颈液基细胞学与组织病理学的对照分析

目的探讨宫颈液基细胞学诊断与组织病理学诊断的符合情况。方法对15393例做SurePath宫颈液基细胞学检查,结果异常者依次做Hybrid Capture-Ⅱ肿瘤相关人乳头状瘤病毒（HPV）-DNA检测、5％醋酸宫颈染色肉眼观察并拍照、阴道镜检查及宫颈多点活检行组织病理学检查。细胞学诊断采用TBS（2001）分级报告系统,阳性诊断包括意义不明的不典型鳞状细胞（ASC-US）以上病变;本组细胞学阳性病例有组织病理诊断结果,并对两者进行了对照分析。结果15393例宫颈细胞学检查与组织病理学对照结果显示：7例鳞状细胞癌（SCC）均符合,高级别鳞状上皮内病变（HSIL）为93．6％（103／110）、低级别鳞状上皮内病变（LSIL）为82．0％（443／540）。HPV—DAN阳性检出率与细胞学TBS分级及组织病理学分级正相关。结论应用液基细胞学制片方法、准确掌握TBS的诊断标准可确保宫颈细胞学检查的准确性。     

传统宫颈细胞学检查与薄层细胞学检测的分析比较与评价

目的探讨宫颈液基细胞学检测系统用于宫颈癌防癌涂片的筛查与传统宫颈涂片检查相比的优越性。方法回顾性分析了1051例TCT普查人群及同时期1050例CS普查人群的宫颈涂片结果。对两种方法的标本满意率及宫颈病变的阳性检出率进行了比较，并对两种方法与组织学的符合率进行了比较。结果TCT组标本满意率明显高于CS组（P〈0．01）；对非典型鳞状上皮细胞及其以上病变的检出率TCT组亦明显高于CS组（P〈0．05）。以阴道镜下组织学活检结果为标准，TCT组与CS组对LSLL及HSIL的诊断符合率分别为86．44％，88．88％及66．7％，83．33％。结论与CS相比，TCT技术明显提高了标本满意率及宫颈病变的阳性检出率，TCT技术是宫颈病变筛查的有效手段。     

液基薄层细胞学检测及TBS分类在宫颈病变诊断中的应用

目的探讨液基薄层细胞学（TCT）检测在宫颈病变诊断中的价值。方法对648例宫颈异常的患者进行液基薄层细胞学检测，将诊断意义不明的不典型细胞（ASCUS）以上病变者行阴道镜下活检，将细胞学检测结果与活检结果作对比分析。结果648例TCT检测的患者中，宫颈病变发生率达81．9％，其中良性病变348例，占53．7％，宫颈上皮内病变183例，占28．2％，对183例异常者进行阴道镜下活检与组织病理学诊断比较，符合率为83．6％。宫颈上皮内病变高发年龄为30～40岁，占37．8％。结论TCT技术在宫颈病变的诊断中，具有简便、实用，准确率高的特点，配合阴道镜检查能及时发现宫颈早期病变，是防止宫颈癌的关键。     

液基薄层细胞学检查与宫颈刮片检查筛查宫颈癌的对比研究

目的 比较宫颈脱落细胞薄层细胞学检查(TCT)和宫颈刮片(CS)检查诊断宫颈病变的准确性。方法 回顾性分析2012年1—12月蚌埠医学院第一附属医院健康体检管理中心对已婚健康妇女进行宫颈脱落细胞检查的资料,其中TCT(TCT组) 787例,CS检查(CS组)11 543例,采用宫颈细胞学诊断报告方式(TBS)作出细胞学诊断,对检查结果异常者行宫颈组织病理学检查,对检查结果进行对比分析。结果 CS组11 543例,宫颈脱落细胞检查阳性105例,阳性检出率0.91%(105/11 543)。TCT组787例,宫颈脱落细胞检查阳性23例,阳性检出率2.92%(23/787)。TCT组的阳性检出率明显高于CS组,差异有统计学意义(χ²=29.056, P＜0.01)。两组宫颈脱落细胞检查结果阳性者共128例,其中41例行宫颈组织病理学检查。41例中,CS组25例,与病理诊断符合11例(44.0%);TCT组16例,与病理诊断符合9例(56.3%);两组病理诊断的符合率差异无统计学意义(P＞0.05)。结论 在宫颈癌的筛查中,TCT的检出率明显高于CS检查。在技术设备和经济条件许可的地区,宫颈癌筛查首选TCT;而在经济发展落后的地区,CS检查仍为宫颈癌筛查的首选。     

薄层液基细胞学联合DNA定量分析检测在肺癌中的诊断价值

目的探讨痰薄层液基细胞学(thin-cytologic test,TCT)和DNA定量分析技术(DNA-image cytometry,DNAICM)联合检测在肺癌中的临床应用价值。方法收集247例肺癌患者痰标本为实验组,另收集247例正常人痰标本作为对照组;采用TCT和DNA-ICM两种方法检测晨痰标本。结果痰DNA-ICM分析的敏感度高于TCT,但特异度低于TCT。TCT在痰标本中的敏感度为49.4%、特异度为95.1%,DNA-ICM分别为72.5%、88.7%,敏感度差异有统计学意义(P 0.01)。TCT与DNA-ICM联合检测诊断阳性为197例,敏感度为79.8%,特异度为84.6%;与TCT相比,敏感度差异有统计学意义(P 0.01)。结论痰TCT结合DNA-ICM联合检测可显著提高肺癌的检出率。     

p16/Ki-67双染联合HC2检测在液基细胞学诊断中的应用

目的 探讨p16/Ki-67细胞学双染联合高危型HPV(HR-HPV)检测在宫颈液基细胞学诊断中的应用价值.方法 选择2017年1月至2019年1月在核工业四一六医院行宫颈液基细胞学(TCT)检查,报告结果为无上皮内病变或恶性病变(NILM)的患者226例,将其剩余保存液行p16/Ki-67免疫细胞化学染色,同时行二代...     

薄层液基细胞学联合P16蛋白检测对宫颈癌前病变的诊断价值

目的:探讨薄层液基细胞学检测（TCT）联合P16免疫细胞染色检查在宫颈癌前病变筛查中的价值。方法:以2019年5月至10月在本院就诊的108例患者为研究对象,收集宫颈脱落细胞,应用免疫组化检测P16表达;应用薄层液基细胞学筛查（TCT）对脱落细胞进行宫颈癌筛查;在阴道镜下对可疑病变区进行多点病理活组织取样,进行病理学诊...     

Characteristics of apparently false-negative digene hybrid capture 2 high-risk HPV DNA testing

This study characterized cases with a negative high-risk Hybrid Capture 2 (HRHC2; Digene, Gaithersburg, MD) test result with concurrent or follow-up biopsy-confirmed high-grade cervical intraepithelial neoplasia (CIN 2/3). From 2,306 HRHC2 tests, 10 negative results were identified with CIN 2/3 (false-negative rate, 4.5%). The majority of the patients had abnormal colposcopic findings and high-grade squamous intraepithelial lesion (HSIL) shown by concurrent cytologic examination, although with few abnormal cells. No trend was evident in the location of the dysplastic epithelium or overall lesion size. In 4 tests, the relative light units over cutoff was more than 0.4 but less than 1.0, suggesting that low quantities of human papillomavirus (HPV) DNA were present in the sample. The negative predictive value for HRHC2 testing may be compromised when the copy number of the HPV DNA is low, and a negative HRHC2 test result may be falsely negative in patients with abnormal colposcopic findings or concurrent cytologic findings showing HSIL.     

Resolution of equivocal results with the Hybrid Capture II high-risk HPV DNA test: a cytologic/histologic review of 191 cases.

INTRODUCTION: The Hybrid Capture II (HC II, Digene) high-risk human papilloma virus (HPV) (hrHPV) DNA test is an in vitro nucleic acid hybridization assay that uses enhanced chemiluminescence for the qualitative and semiquantitative detection of hrHPV in cervical samples. Patient samples are concomitantly tested with positive and negative DNA controls and results reported as positive or negative on the basis of a ratio of relative light units to a cutoff value derived from the positive control (RLU/CO). Samples with a ratio <1.0 RLU/CO are expressed as negative for hrHPV, samples with a ratio >2.5 RLU/CO are expressed as positive for hrHPV, and samples with a ratio between these numbers are submitted for retesting. These "equivocal" values are resulted as positive for hrHPV if either of 2 subsequent test values equals or exceeds 1.0 RLU/CO. Samples that show <1.0 RLU/CO after 2 repeat tests are resulted as negative for hrHPV. METHODS: In this study, we evaluated all hrHPV test results over a 17-month period in our institution. Initial tests showing an equivocal result were analyzed for final retesting result, and for all corresponding and subsequent cytology and histology results. All hrHPV tests were conducted on SurePath (TriPath) or ThinPrep (Cytyc) cervical cytology specimens using the HC II hrHPV DNA test. Subsequent hrHPV tests also were correlated with incident and follow-up findings. RESULTS: A total of 4792 hrHPV test results were evaluated. Of these, 191 (4%) showed equivocal initial results. When retested, 178 of the 191 samples (93%) resulted positive for hrHPV on first retest and an additional 8 resulted positive for hrHPV on the second retest, bringing the total positive tests to 186 out of 191 (97.4%). Five samples (2.6%) out of 191 were finally expressed as negative for hrHPV. Corresponding cytologic interpretations for the 191 specimens were as follows: NILM-30, atypical squamous cell of undetermined significance (ASC-US)-138, atypical squamous cells--cannot exclude HSIL-13 (ASC-H-13), LSIL-9, and high-grade squamous intraepithelial lesion (HSIL)-1. Follow-up histology was available for 60 of the 191 equivocal cases and showed cervical intraepithelial neoplasia (CIN) II or CIN III in 7 cases, CIN I in 13 cases, and negative or reactive changes in 40 cases. CONCLUSIONS: On the basis of the results, repeat testing of equivocal specimens might not be necessary as these specimens are overwhelmingly found to be positive for hrHPV. Additionally, hrHPV tests falling in the equivocal range should be considered as definite positive tests, as follow-up results in this cohort demonstrate that significant histologic abnormalities are associated with 10.5% of these cases (20/191), and with 33% of those biopsied (20/60) cases.     

Detection of HPV DNA in cervical carcinomas by PCR and hybrid capture assay

HPV DNA was detected in exfoliated cervical cells of 73% (85/116) cervical cancer patients by PCR using HPV consensus primers and by hybrid capture assay (HC II) (Digene Corp., USA) in 77 of the 85 cases found HPV positive by PCR. Presence of HPV 16/18 DNA were investigated in the 79 cases by PCR using type specific primers. HPV 16 was detected in 31 (39%) patients, HPV 18 in 7 (8.8%), both HPV 16 and 18 in 19 (24%) and HPVs other than 16/18 in 22 (27.8%) cases. Age and clinical stages had no significant effect on HPV prevalence. Double infection of HPV 16 and 18 was significantly (p<0.05) high in the older patients (56 years or more) compared to younger group. Results indicated that cervical cancers in India are strongly associated with high-risk type HPV infection. HC II assays and PCR results for detection of HPV in cervical smears were comparable.     

Prevalence and typing of HPV DNA by hybrid capture II in women with ASCUS, ASC-H, LSIL, and AGC on ThinPrep Pap tests

Testing for human papillomavirus (HPV) DNA is now a viable option for the management of women with atypical squamous cells of undetermined significance (ASCUS). The utility of reflexive HPV DNA testing for women with a cytologic diagnosis of atypical glandular cells-not otherwise specified (AGC-NOS), ASCUS subtypes, and low-grade squamous intraepithelial lesion (LSIL) has not been well established. In the present investigation, reflex Hybrid Capture II HPV DNA testing results were evaluated for HPV prevalence and type in 371 women with abnormal cytologic diagnoses of ASCUS-not otherwise specified (ASCUS-NOS), ASCUS-suspicious for low-grade squamous intraepithelial lesion (ASCUS-L), atypical squamous cells-cannot exclude high-grade squamous intraepithelial lesion (ASC-H), AGC-NOS, and LSIL on ThinPrep Pap tests. Positive high-risk HPV DNA was identified in 53.6% of the study samples, including ASCUS-NOS 40.2% ASCUS-L 71.4%, ASC-H 37.5%, LSIL 88.6%, and AGC-NOS 0%. We conclude that reflex HPV DNA testing appears to not be useful for colposcopy triage for cytologic diagnoses of LSIL or AGC-NOS.     

Human papillomavirus DNA testing by PCR-ELISA and hybrid capture II from a single cytological specimen: concordance and correlation with cytological results.

BACKGROUND AND OBJECTIVES: A persistent infection by high-risk HPV is now considered as the major cause of cervical carcinoma. The use of a single cytological specimen for HPV DNA testing by two different molecular methods was analyzed and validated. STUDY DESIGN: HPV DNA testing by PCR-ELISA and hybrid capture II HPV test (HC-II), was investigated on 317 cytological samples obtained from Italian women. Two hundred twenty-seven women were referred to virological lab for HPV DNA testing during cytological routine screening and 90 during a cytological and virological follow-up after a conization or hysterectomy. RESULTS: Overall, the concordance between the two assays was high (K=0.87). Compared with PCR-ELISA, the HC-II showed a sensitivity of 91.7% and a specificity of 95.4%. Although the analytical sensitivity of the PCR-ELISA was higher, the performance of the two tests did not differ in recognizing HPV DNA positive patients with either low or high-grade squamous intraepithelial lesions (LSIL or HSIL). HPV DNA positivity was directly correlated with the severity of cytological diagnosis (P<0.005). CONCLUSIONS: In view of the comparable results obtained with the two assays and of the ease of use, and higher throughput of HC-II, it seems advisable, with a single cytological specimen, to employ the HC-II test as a first-line assay, either for screening or diagnosis, and to perform reflex PCR on positive samples, if typing of prevalent high risk HPVs is needed.     

A population-based clinical trial comparing endocervical high-risk HPV testing using hybrid capture 2 and Cervista from the SHENCCAST II Study

Our objective was to directly compare the accuracy of the high-risk human papillomavirus (HPV) assays, Hybrid Capture 2 (hc2; Qiagen, Gaithersburg, MD) and Cervista (Hologic, Bedford, MA), in diagnosing cervical intraepithelial neoplasia (CIN) 3 or worse (cancer). A population-based, cross-sectional study (The Shenzhen Cervical Cancer Screening Trial II) was conducted in Guangdong Province in China. Three high-risk HPV assays, self and direct cervical sampling and cytology, were studied. Abnormal results on any of 6 study tests (33%) resulted in referral to colposcopy. At colposcopy, every patient had at least 5 cervical biopsy specimens obtained. For 8,556 women between the ages of 25 and 59 years (mean, 38.9 years), the rate for CIN 3 or worse was 1.6% (141/8,556). The sensitivity (confidence interval) values for CIN 3 or worse were 97.9% (94.0%-99.6%) and 95.1% (90.0%-98.0%) for hc2 and Cervista, respectively (P > .05). The specificity (confidence interval) values were 87.8% (87.1%-88.5%) and 90.3% (89.6%-90.9%), respectively (P < .05). Differences in accuracy in diagnosing CIN 3 or worse with the hc2 and Cervista tests are minor and result from the decisions made in selecting the cut points.     

Accuracy and interlaboratory reliability of human papillomavirus DNA testing by hybrid capture.

Epidemiologists and clinicians wishing to introduce human papillomavirus (HPV) testing into cervical cancer prevention programs need standardized, reliable, and accurate HPV DNA tests that can detect the full spectrum of pathogenic HPV types. The Hybrid Capture System assay from Digene (hybrid capture assay) is a nonradioactive kit designed to detect 14 HPV types in two groups: a mix of 9 high-risk types associated with anogenital cancer (HPV types 16, 18, 31, 33, 35, 45, 51, 52, and 56) and another group of 5 low-risk types associated with condyloma acuminatum (HPV types 6, 11, 42, 43, and 44). The assay yields quantitative data meant to reflect viral concentration. In a study of 199 cervical specimens from women with concurrent Pap smears, we assessed the reliability of the new assay by comparing the hybrid capture assay results from three laboratories. We assessed the accuracy of the hybrid capture assay in comparison with a reference standard of HPV DNA content (multiple testing by several methods in two reference laboratories). We also compared the hybrid capture assay results with the concurrent cytologic diagnoses on the basis of an independent review of each smear by five pathologists. Pairwise interlaboratory agreement rates on HPV positivity for either high-risk or low-risk types ranged from 87 to 94%, and kappa values ranged from 0.61 to 0.83. Among specimens positive for high-risk types (the most important clinical outcome), the interlaboratory correlations of the quantitative data ranged from 0.60 to 0.90.(ABSTRACT TRUNCATED AT 250 WORDS)     

A novel filtration-based processing method of liquid cytology specimens for human papillomavirus DNA testing by hybrid capture II

We evaluated a more efficient method of processing liquid-based cervical cytology specimens for human papillomavirus (HPV) DNA testing by Hybrid Capture II (HCII). Aliquots were made from 701 specimens in the following sequence: 4.0, 2.0, 1.0, 0.5, and 1.5 mL. The 4.0-mL aliquot was processed by the standard method (STP), and half of the processed material was tested by HCII. Other aliquots were processed with a new, filtration-based processing method (NPM). The 2.0-mL NPM aliquot had HCII test performance most similar to the STP, ie, similar HCII positivity (P = .4) and good test agreement (kappa = 0.85, 95% confidence interval [CI], 0.80-0.89). The 194 cytologic negatives had greater positivity by STP (P = .04) compared with the 2.0-mL aliquot processed by NPM; between-method agreement was modest (kappa = 0.54, 95% CI, 0.36-0.72). A lower positive cut point for the 2.0-mL NPM aliquot partially abrogated this minor difference. In 241 specimens diagnosed as low-grade and 31 as high-grade squamous intraepithelial lesions, there were no significant differences in HPVpositivity (>85% and 90%, respectively) between STP and NPM. NPM reduces specimen handling and decreases total testing time by approximately 33% without significant losses in HCII test performance.     

Fine-needle aspiration biopsy diagnosis of rhabdomyosarcoma: cytologic, histologic, and ultrastructural correlations.

A series of 15 cases of rhabdomyosarcoma diagnosed by fine-needle aspiration biopsy (FNAB) and confirmed by histopathology is reviewed. Cytologically, the tumors were composed of a variable mixture of cells, which according to the degree of differentiation were categorized as early, intermediate, or late rhabdomyoblasts. Histologically, the tumors were divided into embryonal 9, monomorphic round cell 4, and alveolar rhabdomyosarcoma 2. Comparison of histological and cytological features revealed that embryonal types were composed mainly of early rhabdomyoblasts. Recognition of these patterns may be helpful in FNAB diagnosis of rhabdomyosarcoma.     

Comparison of the cobas Human Papillomavirus (HPV) test with the hybrid capture 2 and linear array HPV DNA tests

The cobas human papillomavirus (HPV) test (cobas) was recently approved by the U.S. Food and Drug Administration (FDA) and identifies HPV16 and HPV18 separately as well as detecting a pool of 11 HR-HPV genotypes (HPV31, -33, -35, -39, -45, -51, -52, -56, -58, -59, -68) and also HPV66. We compared cobas, Linear Array (LA), and Hybrid Capture 2 (HC2) assays for detection of carcinogenic HPV DNA, and cobas and LA for detection of HPV16 and HPV18 DNA, among the first 1,852 women enrolled in the HPV Persistence and Progression Cohort (PaP Cohort) study. Specimens were tested by all 3 assays 1 year after an HC2-positive result. In 1,824 specimens with cobas results, cobas had an 85.9% agreement with HC2 and 91.0% agreement with LA for carcinogenic HPV detection. When results between cobas and HC2 disagreed, cobas tended to call more women HPV positive (P < 0.01). Categorizing cobas and LA results hierarchically according to cancer risk (HPV16, HPV18, other carcinogenic HPV genotypes, or carcinogen negative), there was a 90% agreement for all categories of HPV (n = 1,824). We found good agreement between the two U.S. FDA-approved HPV tests, with discrepancies between the two assays due to specific characteristics of the individual assays. Additional studies are needed to compare HC2 and cobas for detecting and predicting CIN3 to understand the clinical implications of the discrepant test results between the two tests.