HPV分型检测在宫颈病变中的诊断价值

目的 探讨人乳头状瘤病毒（Human papillomavirus,HPV）感染状况和基因型分布情况,以及与宫颈病变的关系。方法 选取2016年6月‒2017年6月于青岛市市立医院妇科门诊就诊,有性生活史并要求行HPV亚型检测的患者,共计11 885例,其中5 434例患者同时行液基薄层细胞学检测（thinprep cytologic test,TCT）,分别分析HPV感染率、亚型与宫颈病变的关系。结果 11 885例受检者中HPV总感染率为33.51%（3 983/11 885）,其中高危型HPV占82.59%（5 076/6 146）,主要亚型为HPV16、HPV52、HPV58;低危型HPV占17.41%（1 070/6 146）,主要亚型为HPV81、HPV6、HPV11。HPV 16/18型阳性率随宫颈细胞学病变程度的升高而显著增加（χ2=1 090.12,P<0.001）,HSIL组感染率最高（62.43%）。HSIL组三种最常见的HPV是HPV16（41.76%）、HPV58（11.49%）、HPV33（8.05%）。HPV33阳性率随宫颈细胞学病变程度的升高而显著增加（χ2=13.78,P=0.003）。结论 HPV型别、感染率与宫颈高度鳞状上皮内病变密切相关,为宫颈癌的筛查、防治提供初步理论依据。     

HPV testing for cervical cancer screening in Croatia

Opportunistic screening based on the Pap smear has been undertaken in Croatia since 1953. However, cervical cancer remains an important health problem in Croatia when compared to European countries with organised screening programmes. In Croatia, in addition to screening based on well established cytology, Human papillomavirus (HPV) testing is widely used as secondary test as a triage to borderline cytology and as a follow-up after treatment of severe cervical lesions. Many different approaches for HPV testing arose in Croatia over the last decade depending on the needs of each medical institution involved. Presently, there is an urgent need for better networking between the laboratories, the implementation of quality assessment and the adaptation of a uniform system of referring to and reporting of HPV testing. In conclusion, the best possible organisation for HPV testing would be essential for implementation of HPV testing as primary screening test in Croatia, an thus ultimately and hopefully, the more successful cervical cancer control.     

青岛地区HPV亚型检测及其在宫颈病变中的意义

目的 调查青岛地区妇科门诊就诊女性人群人乳头状瘤病毒（human papillomavirus,HPV）感染状况和基因型分布情况,以及与宫颈病变的关系。方法 选取2016年6月-2017年6月于我院妇科门诊就诊,有性生活史并要求行HPV亚型检测的患者,共计11 885例,其中5 434例患者同时行TCT检测,分别分析年龄、多重感染及HPV亚型与宫颈病变的关系。结果 11 885例受检者中HPV总感染率为33.51%（3 983/11 885）,其中：高危型HPV占82.59%（5 076/6 146）,主要包括HPV16、HPV52、HPV58;低危型HPV占17.41%（1 070/6 146）,主要包括HPV81、HPV6、HPV11。HPV单一亚型感染率为63.65%（2 535/3 983）,多重感染率为36.35%（1 448/3 983）。不同年龄段HPV感染率差异有统计学意义（P＜0.01）,呈“U”型,≤29岁感染率最高（43.20%）,HPV感染率随TCT检查诊断级别的升高而显著升高（P＜0.01）,HSIL组HPV感染率最高（93.12%）。结论 青岛地区高危型HPV主要亚型为HPV16、HPV52、HPV58;不同年龄段及不同TCT诊断级别HPV感染率不同,HPV感染与宫颈鳞状上皮内病变密切相关,为宫颈癌的筛查、防治提供初步理论依据。     

HPV genotypes in high grade cervical lesions and invasive cervical carcinoma as detected by two commercial DNA assays, North Carolina, 2001-2006

Background

HPV typing using formalin fixed paraffin embedded (FFPE) cervical tissue is used to evaluate HPV vaccine impact, but DNA yield and quality in FFPE specimens can negatively affect test results. This study aimed to evaluate 2 commercial assays for HPV detection and typing using FFPE cervical specimens.

Methods

Four large North Carolina pathology laboratories provided FFPE specimens from 299 women ages18 and older diagnosed with cervical disease from 2001 to 2006. For each woman, one diagnostic block was selected and unstained serial sections were prepared for DNA typing. Extracts from samples with residual lesion were used to detect and type HPV using parallel and serial testing algorithms with the Linear Array and LiPA HPV genotyping assays.

Findings

LA and LiPA concordance was 0.61 for detecting any high-risk (HR) and 0.20 for detecting any low-risk (LR) types, with significant differences in marginal proportions for HPV16, 51, 52, and any HR types. Discordant results were most often LiPA-positive, LA-negative. The parallel algorithm yielded the highest prevalence of any HPV type (95.7%). HR type prevalence was similar using parallel (93.1%) and serial (92.1%) approaches. HPV16, 33, and 52 prevalence was slightly lower using the serial algorithm, but the median number of HR types per woman (1) did not differ by algorithm. Using the serial algorithm, HPV DNA was detected in >85% of invasive and >95% of pre-invasive lesions. The most common type was HPV16, followed by 52, 18, 31, 33, and 35; HPV16/18 was detected in 56.5% of specimens. Multiple HPV types were more common in lower grade lesions.

Conclusions

We developed an efficient algorithm for testing and reporting results of two commercial assays for HPV detection and typing in FFPE specimens, and describe HPV type distribution in pre-invasive and invasive cervical lesions in a state-based sample prior to HPV vaccine introduction.     

Evaluation of a cervical cancer screening program based on HPV testing and LLETZ excision in a low resource setting

We conducted studies in Vanuatu to evaluate potential screening and treatment strategies to assist with control of cervical cancer. In a pilot study of 496 women, visual inspection and cytology were evaluated as screening tests for detection of CIN 2 or worse (CIN2+), observed in 21 of 206 subjects biopsied on the basis of abnormal visual inspection or cytology. Sensitivity of visual inspection with Lugol's Iodine for detection of CIN2+ on biopsy was 0.63, specificity was 0.32, and the positive predictive value was 0.09. For HSIL cytology, sensitivity was 0.99, specificity was 0.77, and the positive predictive value was 0.88. HSIL cytology was significantly more sensitive and had a significantly higher PPV for CIN 2+ than visual inspection (p<0.01). In a further study of 514 women, we compared testing for HR HPV and cytology as predictors of biopsy proven CIN 2+. Sensitivity of HSIL cytology for CIN2+ as established by loop excision of the cervix was 0.81, specificity was 0.94, and positive predictive value was 0.48. Sensitivity of a positive test for HR HPV for detection of CIN2+ was non-significantly different from cytology at 0.81, specificity was 0.94, and positive predictive value was 0.42. Combining the two tests gave a significantly lower sensitivity of 0.63, a specificity of 0.98, and a positive predictive value of 0.68. For women over 30 in a low resource setting without access to cytology, a single locally conducted test for high risk HPV with effective intervention could reduce cervical cancer risk as effectively as intervention based on cytology conducted in an accredited laboratory.     

HPV E6/E7 mRNA testing is more specific than cytology in post-colposcopy follow-up of women with negative cervical biopsy

Background

In Norway, women with negative or low-grade cervical biopsies (normal/CIN1) are followed up after six months in order to decide on further follow-up or recall for screening at three-year intervals. A high specificity and positive predictive value (PPV) of the triage test is important to avoid unnecessary diagnostic and therapeutic procedures whereas a low risk of high-grade disease among triage negative women assures safety.

Materials and Methods

At the University Hospital of North Norway, cytology and the HPV mRNA test PreTect HPV-Proofer, detecting E6/E7 mRNA from HPV types 16, 18, 31, 33 and 45, are used in post-colposcopy follow-up of women with negative or low-grade biopsy. In this study, women with negative biopsy after high grade cytology (ASC-H/HSIL) and/or positive HPV mRNA test in the period 2005–2009 were included (n = 520). Histologically confirmed cervical intraepithelial neoplasia of grade 2 or worse (CIN2+) was used as study endpoint.

Results

Of 520 women with negative or low-grade biopsy, 124 women (23.8%) had CIN2+ in follow-up biopsy. The sensitivity and specificity of the HPV mRNA test were 89.1% (95% CI, 80.1–98.1) and 92.5% (95% CI, 88.2–96.7), respectively. The ratios of sensitivity, specificity and PPV of HPV mRNA testing compared to repeat cytology for finding CIN2+ was 1.05 (95% CI: 0.92–1.21), 1.21 (95% CI: 1.12–1.32), and 1.49 (95% CI: 1.20–1.86), respectively. The PPV of mRNA was 77.3% (95% CI, 59.8–94.8) in women aged 40 or older.

Conclusion

Women with negative cervical biopsy require follow-up before resumption of routine screening. Post-colposcopy HPV mRNA testing was as sensitive but more specific than post-colposcopy cytology. In addition, the HPV mRNA test showed higher PPV. A positive mRNA test post-colposcopy could justify treatment in women above 40 years.     

HPV16感染检测和TCT筛查联用的临床应用

目的评估人乳头瘤病毒(Human papillomavirus,HPV)16感染和液基薄层细胞学(thinprep cytologic test,TCT)筛查在预测宫颈病变中的临床价值。方法以门诊537例高危型HPV感染疑似宫颈病变女性为对象,进行HPV16感染分析、液基薄层细胞学和阴道镜病理检查,以病理活检为金标准,比较HPV16感染筛查方法、TCT筛查方法以及二者联用在筛查中的敏感度、特异度等指标,判断其临床应用价值。结果 HPV16感染筛查方法的敏感度为62.9%,特异度为83.5%,阳性预测值为53.4%,阴性预测值为88.2%;TCT筛查方法的敏感度为41.2%,特异度为92.7%,阳性预测值为62.9%,阴性预测值为84.0%;二者联合筛查,以HPV16感染或TCT异常为阳性,敏感度为84.7%,特异度为75.8%,阳性预测值为51.2%,阴性预测值为94.3%。结论 HPV16感染联合TCT异常筛查可以提高筛查的灵敏度,特异度也在可接受范围内,可以作为宫颈防癌初筛方法。     

HPV分型及其高危亚型与宫颈病变的关系

目的研究HPV分型及其高危亚型与宫颈病变程度的关系。方法采用HPV基因分型技术对2014年7月至2015年8月在广州中医药大学第一附属医院妇科门诊就诊和体检的3 142例患者进行HPV分型检测,其中327例患者进行宫颈病理检查,分析高危型HPV感染与宫颈病变程度的关系。结果 3 142例受检对象中,21种HPV型别均被检测出,阳性检出者共969例,总检出率为30.80%,其中高危型741例,检出率为23.58%。随着宫颈病变级别的升高,高危型HPV阳性率也相应升高(P0.05),HPV16型感染阳性者宫颈癌发生率最高(P0.05)。高危型HPV阳性者中单一感染和多重感染与宫颈病变程度无关(P0.05)。结论高危型HPV亚型检测在宫颈病变预防和诊治中具有重要意义。     

Logistic regression analysis of low grade spindle cell lesions. A cytologic study

OBJECTIVE: To identify key diagnostic cytologic criteria for various low grade spindle cell lesions. STUDY DESIGN: We reviewed 20 synovial sarcomas, 18 benign neural tumors, 10 reparative lesions, 24 other benign and 27 additional malignant low grade spindle cell lesions. All specimens were coded as to the presence or absence of the following variables: high cellularity, tissue fragments, tissue culture appearance, epithelial fragments, vessel fragments, vascular arcades, fibrillar ground substance, myxoid background, microcystic areas, parallel arrangement of nuclei, naked nuclei, single cells, binucleate cells, multinucleate cells, long filamentous cells, short spindle cells, stellate cells, lipoblasts, nuclear pleomorphism, nuclei with pointed ends, comma/fishhook nuclei, cigar-shaped nuclei, ovoid/round nuclei, small nucleoli, large nucleoli, mitotic figures, intranuclear vacuoles and background histiocytes. A logistic regression analysis was performed to identify the variables predictive of malignant lesions, specifically synovial sarcomas, benign neural tumors and reparative lesions. RESULTS: Statistical analysis selected high cellularity, short spindle cells, small nucleoli and absence of tissue culture appearance as the main criteria for malignant neoplasms. Tissue fragments and high cellularity were selected as the primary criteria and absence of long filamentous cells and of myxoid background as the secondary criteria for synovial sarcomas. It selected fibrillar ground substance and absence of ovoid/round nuclei as the key criteria for benign neural tumors. The presence of a tissue culture appearance was the major criterion for reparative lesions. CONCLUSION: There are many previously described cytologic criteria, but we found that when subjected to statistical analysis, only a few features were significant in the evaluation of low grade spindle cell lesions.     

Combined Pap smear, cervicography and HPV DNA testing in the detection of cervical intraepithelial neoplasia and cancer

OBJECTIVE: The sensitivity of the Pap smear (PAP) continues to be the subject of debate. During the past several years, cervicography (CER) and HPV DNA testing have been suggested as optional tools in the screening of cervical cancer precursors. STUDY DESIGN: The performance characteristics of PAP, CER and HPV DNA testing (hybrid capture test [HCT]) in all potential combinations were evaluated in a series of 1,030 women (aged 16-70, median, 33), subjected to colposcopy (COLPO) as the reference tool. RESULTS: Of the 992 evaluable cases, 402/992 (41%) had positive COLPO (i.e., an abnormal transformation zone). Of them, 298 women underwent directed punch biopsy, while of the COLPO negative patients, 18/93 positive by at least one of the three tests had endocervical curettage. Of the 402 COLPO positive women, 146 (36%) remained negative on all tests, whereas 256 (64%) had at least one positive test. There were 84 cervical intraepithelial neoplasia (CIN) 2 and 3 lesions and 6 invasive carcinomas. Of the former, 10 were detected by PAP alone, 4 by CER alone and 3 by HCT alone. Three of the 6 carcinomas were HCT negative. The predictive value (PPV) of a positive test was 45% for PAP, 51% for CER and 48% for HCT. The combinations of PAP with CER (for PAP negative cases) and PAP with HCT were more sensitive for CIN 2 and 3 (95% and 94%, respectively) as compared with PAP alone but were associated with a significant decrease in specificity (44% and 46% vs. 57%, respectively). However, both combinations retained a PPV (43%) similar to that of PAP alone (45%). CONCLUSION: The potential combinations of PAP with CER and with HCT were more sensitive in detecting CIN 2 and 3 as compared with PAP alone and retained a PPV similar to that of PAP.     

Relationship of up-regulation of 67-kd laminin receptor to grade of cervical intraepithelial neoplasia and to high-risk HPV types and prognosis in cervical cancer

OBJECTIVE: To evaluate the 67-kd laminin receptor (67LR) in cervical cancer and its molecular links to oncogenic HPV types. STUDY DESIGN: As part of the HPV-PathogenlSS Study, a series of 150 squamous cell carcinomas (SCCs) and 152 carcinoma in situ (CIN) lesions were examined using immunohistochemical staining for LR67 and tested for HPV using polymerase chain reaction (PCR) with 3 primer sets (MY09/11, GP5+/GP6+, SPF). Followup data were available for all SCC patients, and 67 CIN lesions had been monitored with serial PCR for HPV clearance/persistence after cone treatment. RESULTS: 67LR expression increased in parallel with increasing grade of CIN (p = 0. 0001), with the most dramatic up-regulation upon the transition from CIN 2 to CIN 3 and further to SCC. This increased expression was associated with CIN 3/cancer at OR 17.04 (95% CI 7.28-39.87). The seemingly significant association of 67LR with high-risk HPV (HR-HPV) detection (OR 2.20, 95% CI 1.27-3.80) was due to confounding by the histologic grade (Mantel-Haenszel common OR = 1.118, 95% CI 0.576-2.168). Using performance indicators, 67LR expression was of little value as a marker of HR-HPV type, and it did not predict clearance/persistence of HR-HPV after treatment of CIN. Similarly, 67LR expression was not an independent prognostic factor in cervical cancer. CONCLUSION: In cervical carcinogenesis, both integrin- and nonintegrin-type LRs (67LR) probably have functions complementary to each other, mediating transient early and stable adhesions, respectively. Up-regulated 67LR expression is significantly associated with progression from CIN 2 to CIN 3 as a marker of cell proliferation. 67LR is probably orchestrated by mechanisms independent of HR-HPV oncoproteins, which seem to be more closely associated with integrin-type laminin receptors.     

Prediction of recurrent disease by cytology and HPV testing after treatment of cervical intraepithelial neoplasia

Objective: To assess the role of human papillomavirus (HPV) testing and cytology as predictors of residual/recurrent disease after treatment of high‐grade cervical intraepithelial lesions. Methods: One hundred and thirty‐eight women with cervical intraepithelial neoplasia (CIN) grade 2/3 lesion on biopsy were included in a prospective follow‐up study in Belgium and Nicaragua. All women were treated with loop electrosurgical excision procedure (LEEP) and follow‐up visits took place at 6 weeks, 6 months, 1 year and 2 years. During these visits, a Papanicolaou (Pap) smear test was taken, colposcopy was performed and specimens were collected for HPV testing. Cytology, high‐risk (HR) HPV presence, persistent HR HPV infection and combinations of these tests at different time points during follow‐up were correlated with histologically confirmed residual/recurrent disease. Results: Thirteen patients (9%) developed residual/recurrent disease during follow‐up. Abnormal cytology at 6 weeks after treatment was significantly correlated with residual/recurrent disease. Nine of thirty‐seven patients with abnormal cytology at 6 weeks had recurrent disease versus three of seventy with a normal cytology [odds ratio (OR): 7.2; 95% confidence interval (CI): 1.8–28.5; P = 0.003). Sensitivity of this test was 75.0%, specificity 70.5%. Combining abnormal cytology and the presence of HR HPV within the first 6 months after treatment gave the best correlation with residual/recurrent disease: of the 54 women with abnormal cytology and/or HR HPV presence within the first 6 months, 11 developed residual/recurrent disease (OR 10.2; 95% CI: 2.2–48.3). Sensitivity of this combination was 84.6% and specificity 65.0%. Conclusion: Cytology remains the cornerstone in the early follow‐up after LEEP for CIN lesions of the cervix. HPV testing can add value as it increases the sensitivity of cytology in concomitant testing within the first 6 months.