HPV16/18 genotyping for the triage of HPV positive women in primary cervical cancer screening in Chile

Background

We previously conducted a population-based screening trial of high-risk human papillomavirus (hrHPV) testing and conventional cytology, demonstrating higher sensitivity (92.7 % vs 22.1 % for CIN2+) but lower positive predictive value (10.5 % vs 23.9 %) of hrHPV testing. Here we report the performance of HPV16/18 genotyping to triage the hrHPV positive participants.

Methods

Women aged 25 years and older received hrHPV (Hybrid Capture 2) and Papanicolaou testing; positives by either test underwent colposcopy and directed biopsy, as did a sample of double-negatives. hrHPV positive women were reflex-tested with HPV16/18 genotyping (Digene HPV Genotyping PS Test).

Results

Among the 8,265 participants, 10.7 % were hrHPV positive, 1.7 % had ASCUS+ cytology, 1.2 % had CIN2+; 776 (88 %) hrHPV positive women had complete results, of whom 38.8 % were positive for HPV16 (24.0 %), HPV18 (9.7 %) or both (5.1 %). CIN2+ prevalence in HPV16/18 positive women (16.3 %, 95 % CI 12.3-20.9) was twice that of HPV16/18 negative women (8.0 %, 95 % CI 5.7-10.8). HPV16/18 genotyping identified 40.5 % of CIN2, 66.7 % of CIN3 and 75.0 % of cancers. Compared to hrHPV screening alone, HPV16/18 triage significantly reduced the referral rate (10.7 % vs 3.7 %) and the number of colposcopies required to detect one CIN2+ (9 vs 6). When HPV16/18 negative women with baseline ASCUS+ cytology were also colposcopied, an additional 14 % of CIN2+ was identified; referral increased slightly to 4.2 %.

Conclusions

HPV16/18 triage effectively stratified hrHPV positive women by their risk of high-grade lesions. HPV16/18 positive women must be referred immediately; referral could be deferred in HPV16/18 negative women given the slower progression of non-HPV16/18 lesions, however, they will require active follow-up.

     

子宫颈癌患者人乳头瘤病毒16、18检测

 目的　探讨人乳头瘤病毒（HPV）16、18型在子宫颈癌的发生、发展中的意义。方法　应用实时荧光定量聚合酶链反应技术对子宫颈原位癌13例，子宫颈癌Ⅰ期32例，子宫颈上皮内瘤样病变（CIN）Ⅰ~Ⅱ级12例，对照组54例（慢性子宫颈炎组37例、非研究疾病组17例）进行HPV16、HPV18型荧光基因定量检测，计算出HPV DNA的拷贝数。结果　研究组、对照组HPV16、HPV18的感染率差异有统计学意义（P＜0.05），在子宫颈癌发生的不同阶段，HPV16、HPV18的感染定量差异亦有统计学意义。子宫颈癌HPV含量与肿瘤直径大小、浸润间质深度、淋巴结阳性个数无相关性（r = 0.168， r = 0.280， r = 0.333，P＞0.05）；局部肿瘤的直径大小与浸润间质深度呈正相关（r = 0.473，P＜0.05）。结论　致癌性HPV的持续、高浓度存在是子宫颈癌发生、发展的主要因素之一。     

Health and economic impact of HPV 16 and 18 vaccination and cervical cancer screening in India

Cervical cancer is a leading cause of cancer death among women in low-income countries, with approximately 25% of cases worldwide occurring in India. We estimated the potential health and economic impact of different cervical cancer prevention strategies. After empirically calibrating a cervical cancer model to country-specific epidemiologic data, we projected cancer incidence, life expectancy, and lifetime costs (I$2005), and calculated incremental cost-effectiveness ratios (I$/YLS) for the following strategies: pre-adolescent vaccination of girls before age 12, screening of women over age 30, and combined vaccination and screening. Screening differed by test (cytology, visual inspection, HPV DNA testing), number of clinical visits (1, 2 or 3), frequency (1 x , 2 x , 3 x per lifetime), and age range (35-45). Vaccine efficacy, coverage, and costs were varied in sensitivity analyses. Assuming 70% coverage, mean reduction in lifetime cancer risk was 44% (range, 28-57%) with HPV 16,18 vaccination alone, and 21-33% with screening three times per lifetime. Combining vaccination and screening three times per lifetime provided a mean reduction of 56% (vaccination plus 3-visit conventional cytology) to 63% (vaccination plus 2-visit HPV DNA testing). At a cost per vaccinated girl of I$10 (per dose cost of $2), pre-adolescent vaccination followed by screening three times per lifetime using either VIA or HPV DNA testing, would be considered cost-effective using the country's per capita gross domestic product (I$3452) as a threshold. In India, if high coverage of pre-adolescent girls with a low-cost HPV vaccine that provides long-term protection is achievable, vaccination followed by screening three times per lifetime is expected to reduce cancer deaths by half, and be cost-effective.     

Health and economic impact of HPV 16/18 vaccination and cervical cancer screening in Eastern Africa

Eastern Africa has the world's highest cervical cancer incidence and mortality rates. We used epidemiologic data from Kenya, Mozambique, Tanzania, Uganda, and Zimbabwe to develop models of HPV-related infection and disease. For each country, we assessed HPV vaccination of girls before age 12 followed by screening with HPV DNA testing once, twice, or three times per lifetime (at ages 35, 40, 45). For women over age 30, we assessed only screening (with HPV DNA testing up to three times per lifetime or VIA at age 35). Assuming no waning immunity, mean reduction in lifetime cancer risk associated with vaccination ranged from 36 to 45%, and vaccination followed by screening once per lifetime at age 35 with HPV DNA testing ranged from 43 to 51%. For both younger and older women, the most effective screening strategy was HPV DNA testing three times per lifetime. Provided the cost per vaccinated girl was less than I$10 (I$2 per dose), vaccination had an incremental cost-effectiveness ratio [I$ (international dollars)/year of life saved (YLS)] less than the country-specific per capita GDP, a commonly cited heuristic for "very cost-effective" interventions. If the cost per vaccinated girl was between I$10 (I$2 per dose) and I$25 (I$5 per dose), vaccination followed by HPV DNA testing would save the most lives and would be considered good value for public health dollars. These results should be used to catalyze design and evaluation of HPV vaccine delivery and screening programs, and contribute to a dialogue on financing HPV vaccination in poor countries.     

Telomerase assay and HPV 16/18 typing as adjunct to conventional cytological cervical cancer screening.

OBJECTIVE: This study aims at exploring the potential use of telomerase activity assay and typing of human papillomaviruses (HPV) 16 and 18 in improving the identification of high-grade cervical intraepithelial neoplasia (CIN). METHODS: From 86 women with normal cervical smears and from 114 patients with abnormal cervical smears cervical scrapings were collected. The telomerase activity was assayed using the Telomerase Repeat Amplification Protocol, and HPV was detected using consensus primers and specific primers for HPV 16 and HPV 18. RESULTS: HPV 16 in cervical scrapes was significantly associated with high-grade squamous epithelial lesions on cytology and with high-grade CIN, i.e., CIN 2/3 on biopsy. The detection of HPV 18 or telomerase activity had no significant association with high-grade squamous intraepithelial lesions or high-grade CIN. CONCLUSION: The use of the telomerase activity assay in cervical scrapes, unlike HPV 16 typing, did not improve the detection of high-grade CIN.     

Clinical evaluation of human papillomavirus 16/18 oncoprotein test for cervical cancer screening and HPV positive women triage

The prognostic role of tumor‐infiltrating tryptase⁺ mast cells in human solid tumors remains controversial. Herein, we conducted a meta‐analysis including 28 published studies with 4224 patients identified from PubMed and EBSCO to assess the prognostic impact of tumor‐infiltrating tryptase⁺ mast cells in human solid tumors. We found that tryptase⁺ mast cell infiltration significantly decreased overall survival (OS) and disease‐free survival (DFS) in all types of solid tumors. In stratified analyses, tryptase⁺ mast cell infiltration was significantly associated with worse OS in non‐small cell lung cancer, hepatocellular carcinoma and 5‐year survival in colorectal cancer. And these cells were inversely associated with DFS in hepatocellular and colorectal cancer. In addition, high density of intratumoral tryptase⁺ mast cells significantly correlated with lymph node metastasis of solid tumor. In conclusion, Tryptase⁺ mast cell infiltration leads to an unfavorable clinical outcome in solid tumors, implicating that it is a valuable biomarker for prognostic prediction for human solid malignances and targeting it may have a potential for effective treatment.     

Older women testing positive for HPV16/18 on cervical screening and risk of high-grade cervical abnormality

In Australia's HPV-based cervical screening program, we previously showed that risk of histological high-grade abnormality at 1 year post screening decreased with age in women with oncogenic HPV. In this study, we followed 878 HPV16/18 positive women aged 55 years and over for up to 3 years post screening test, to determine the proportion with histological high-grade abnormality (HGA, incorporating high-grade squamous intraepithelial abnormality (HSIL), adenocarcinoma in situ (AIS), squamous cell carcinoma (SCC) and adenocarcinoma) and to correlate risk of HGA with liquid-based cytology result and with prior screening history. HGA was detected in 7.8% at 1 year and 10.0% at 3 years, with no significant difference (P = .136), despite the number of women with follow-up information significantly increasing from 82.9% to 91.0% (P < .0001). The proportion of HPV16/18 positive women with HGA at 3 years was highest in those with an HSIL cytology result (79.0%) and lowest in those with negative cytology (6.2%). Women with an adequate screening history had fewer HGA than such women with inadequate prior screening (6.6% vs 16.0%, P = .001) or with a history of an abnormality (6.6% vs 14.4%, P = .001). HPV16/18 infection in women over 55 years may have a different natural history from that in younger women, in whom HGA are more common after HPV16/18 detection. In HPV-based cervical screening programs, management algorithms for screen-detected abnormalities based on risk stratification should include factors such as age, screening history and index cytology result, so that women receive appropriate investigation and follow-up.     

新柏式薄层液基细胞学(TCT)联合HPV-DNA分型检测在宫颈癌筛查中的临床价值

目的:探讨新柏式薄层液基细胞学（TCT）联合HPV-DNA分型检测在宫颈癌筛查中的临床价值。方法:随机选取2011年4月-2014年4月于我院进行宫颈癌筛查的420例患者为研究对象。所有患者均进行TCT及HPV-DNA筛查,并同时进行宫颈组织活检进行最终确认。结果:TCT筛查结果显示:ASCUS（意义不明的不典型鳞状细胞）患者为175例,阳性比例为41.7%。病理组织筛查结果显示:CINⅠ为199例,阳性比例为47.4%。HPV-DNA共检出阳性患者180例,比例42.9%。临床症状越严重,HPV-DNA阳性率越高。TCT联合HPV-DNA筛查结果灵敏度较单独TCT和HPV-DNA筛查结果高,但特异性低,其阳性预测值和阴性预测值都较高,与病理结果筛查结果更为相似。结论:新柏式薄层液基细胞学(TCT)联合HPV-DNA分型检测在宫颈癌筛查中灵敏度高和准确性较高。     

Human papillomavirus type-distribution in cervical cancer in China: the importance of HPV 16 and 18

Prophylactic vaccination against HPV 16 and 18 has the potential for effective prevention of high-grade precancer (cervical intraepithelial neoplasia [CIN)] 2/3) and ICC caused by these viruses (globally 50 and 70%, respectively) when employed in women prior to starting sexual activity. To provide data for decisions on HPV vaccination in China, we determined HPV type-distribution in ICC and CIN 2/3 from women of different regions within China. A multicenter study was conducted by randomized sampling of paraffin blocks of 664 ICC (630 squamous cell carcinoma [SCC]; 34 adenocarcinoma [ADC]), 569 CIN 2/3 cases from seven regions of China. Histological diagnosis was confirmed in 1,233 cases by consensus review. HPV DNA was detected using the SPF10 LiPA25 version 1 assay. HPV prevalence was 97.6% in SCC, 85.3% in adenocarcinoma, and 98.9% in CIN 2/3. HPV 16 (76.7%) and HPV 18 (7.8%) were the most common, together accounting for 84.5% of SCC, followed by HPV 31 (3.2%), HPV 52 (2.2%), and HPV 58 (2.2%). HPV positivity in SCC did not differ notably by region. However, SCC cases from women ≤34 years had higher HPV 16 positivity than women over 50 years, among whom HPV 52, 58, and 39 were more common. HPV 16 and 18 were under-represented, whereas HPV 31, 52, and 58 were over-represented in CIN2/3 compared to SCC. The potential impact of vaccines against oncogenic HPV types 16 and 18 is estimated to be high (84.5%) against total SCC. These data are critical for China’s future evaluation of the cost-effectiveness of current cervical cancer vaccines and of HPV-based screening guidelines.     

HLA-DR 抗原及HPV16/18E6蛋白在宫颈癌的表达及其相互关系

目的:通过检测HLA-DR抗原及HPV16/18E6蛋白在宫颈癌组织中的表达情况,探讨由HLA-DR抗原介导的免疫应答在感染高危型HPV至发展为宫颈癌过程中的作用机制.方法:采用免疫组织化学SP法检测宫颈癌、CIN及正常宫颈组织中HLA-DR抗原及HPV16/18E6的表达情况.结果:HLA-DR抗原的阳性表达率在宫颈癌、CIN、正常宫颈组织中分别为 81.8%、73.3%、37.5%,而HPV16/18E6蛋白的阳性表达率则分别为75.8%、60.0%、37.5%,两者在三组中的表达均有显著差异 (P均﹤0.05).HLA-DR抗原及HPV16/18E6蛋白在宫颈癌组织中的阳性表达率随临床分期、分化程度变化及淋巴结是否转移而不同,但其差异无显著意义 (P均﹥0.05).结论:HLA-DR抗原递呈病毒抗原给T细胞引起的免疫应答可能在宫颈癌的发生发展过程中起着重要的作用.     

HPV分型研究在宫颈癌筛查中的意义

目的:探讨HPV分型流行病学特征及其与宫颈癌前病变的关系。方法:选择陕西省人民医院2014年1月-2014年12月在妇科门诊就诊、有性生活史并行宫颈液基细胞学（TCT）检查的患者10 885例,其中2 677例患者同时行宫颈感染人乳头瘤病毒（HPV）分型筛查。结果:TCT异常率（≥ASCUS）7.6%; HPV感染率 34.8%,其中高危型HPV占76.93%,低危型HPV占23.93%;混合感染（2种以上HPV亚型感染）占20.28%,高危型HPV感染主要型别为HPV16、HPV52、HPV58;低危型HPV感染主要型别为HPV6、HPV11、HPV43;未发现HPV26、73、83型阳性病例;≤29岁及≥50岁年龄段为HPV感染及TCT异常的高峰年龄段;HPV 感染率随着细胞学诊断级别及病理学诊断级别的升高而显著上升。结论:不同年龄段HPV 分型感染率及 TCT 异常率不同,HPV 感染率与宫颈病变严重程度呈显著正相关。     

Correlation between load of HPV 16 DNA in cervical cancer and HPV 16 DNA in lymph nodes

OBJECTIVE To determine the association between viral load of human papillomavirus 16 （HPV16） DNA in the primary focus of cervical carcinoma and HPV16 DNA in pelvic lymph nodes. METHODS The HPV16 DNA load was measured by fluorescent quantitation polymerase chain reaction （FQ-PCR） in 17 primary foci. HPV16 DNA was detected by polymerase chain reaction （PCR） using HPV16 type-specific primers in 296 pelvic lymph nodes which were from 17 cases of cervical cancer. RESULTS The viral load of HPV16 DNA showed statistically significant differences between tumors with a diameter of 〈 4 cm and ≥ 4 cm （P 〈 0.05）. Seven of 17 cervical cancer cases had HPV16 DNA positive lymph nodes, designated as the positive group, while the remaining 10 without positive lymph nodes was designated the negative group. The average load of HPV16 DNA showed no significant difference between the 2 groups （P 〉 0.05）. The load of HPV16 in the primary lesion was not associated with that in the lymph nodes. There were 38 HPV16 DNA positive nodes in the total 296 nodes. The rate of positivity of HPV16 DNA in lymph nodes showed statistically significant differences in consideration of maximum tumor diameter, tumor differentiation, histologic type, depth of myometial infiltration and the metastatic status of the nodes, respectively （P 〈 0.05）. CONCLUSION Viral load of HPV16 in the primary cancer focus correlated with the quantity of tumor cells in the primary focus but not with the existence of HPV DNA positive lymph nodes. Detection of HPV DNA may help to find the early metastases that cannot be evaluated histopathologically, but the prognostic value of HPV positive lymph nodes needs further examination.     

子宫颈癌患者可溶性尿激酶受体、鳞状细胞癌抗原、人乳头瘤病毒16、人乳头瘤病毒18联合检测的意义

目的 探讨可溶性尿激酶受体（suPAR）、鳞状细胞癌抗原（SCC-Ag）、人乳头瘤病毒（HPV） 16、HPV18联合检测在子宫颈癌患者病情监测及预后评估中的意义.方法 选择206例子宫颈癌患者及60名健康对照者血液和子宫颈分泌物标本,通过酶联免疫吸附试验（ELISA）检测血浆suPAR及SCC-Ag水平,用荧光定量反转录聚合酶链反应法检测子宫颈分泌物HPV16、HPV18的表达,并分析三者的相关性.结果 子宫颈浸润癌患者血浆suPAR、SCC-Ag水平均比健康对照组升高[（1.072 5±0.305 2）ng/ml比（0.501 7 ±0.179 3）ng/ml、（0.980 6±0.162 7）μg/ml比（0.261 4±0.006 3）μg/ml],差异有统计学意义（均P＜ 0.05）;子宫颈浸润癌患者及原位癌患者分泌物HPV16、HPV18阳性表达率分别为53.89％（90/167）和46.15％（18/39）,比健康对照组（6.67％,4/60）升高,差异有统计学意义（P＜0.05）.子宫颈浸润癌患者血浆suPAR水平与SCC-Ag水平呈正相关（r=0.564,P＜0.05）,子宫颈浸润癌患者子宫颈分泌物HPV16、HPV18表达阳性组和阴性组的血浆suPAR水平差异无统计学意义（P＞0.05）.结论 子宫颈浸润癌患者血浆suPAR水平与SCC-Ag水平存在正相关关系,尚不能认为子宫颈浸润癌患者血浆suPAR水平与感染HPV16、HPV18有关.     

早期宫颈癌前哨淋巴结中高危型HPV16/18 DNA表达的检测及临床意义

目的：探讨早期宫颈癌患者前哨淋巴结（SLN）中人乳头状瘤病毒（HPV）16/18 DNA表达检测对于微转移的临床意义。方法选取72例早期宫颈癌患者,予患者均行广泛性子宫切除加双侧盆腔淋巴结清扫术,术中采用染料法识别SLN的宫颈癌患者有46例,应用基因检测法（FQ-PCR）检测SLN中HPV16/18 DNA阳性表达情况,并分析其与各种临床病理因素的关系;对SLN病理阴性的33例患者进行长期随访,分析SLN中HPV16/18 DNA阳性与淋巴结转移的关系。结果46例宫颈癌患者SLN中HPV16/18 DNA阳性表达者共22例,其中13例淋巴结病理阳性患者中有10例阳性,而33例淋巴结病理阴性患者中仅12例阳性（P=0.013）;46例患者共检出前哨淋巴结102枚,均用FQ-PCR法检测HPV16/18 DNA,结果13例淋巴结病理阳性患者检出的37枚SLN中有29枚HPV16/18 DNA阳性,而33例淋巴结病理阴性患者检出的65枚SLN中仅有36枚阳性（P=0.033）;分析46例成功检出SLN的早期宫颈癌患者的临床资料,发现SLN中HPV16/18 DNA阳性表达仅与临床分期有关,具有统计学意义（P=0.034）;长期随访33例SLN病理阴性的患者,发现HPV16/18 DNA阳性的患者复发率高于HPV16/18 DNA阴性的患者,具有统计学意义（P=0.02）。结论检测宫颈癌SLN组织中HPV16/18 DNA表达可能是预测早期宫颈癌淋巴结微转移的可行方法。     

Absence of HPV 16 and 18 DNA in breast cancer.

The finding that human papillomavirus (HPV) genes can immortalise breast epithelial cells has led to suggestions that HPV could be involved in the pathogenesis of breast cancer. Using the polymerase chain reaction (PCR) we have been unable to demonstrate the presence of HPV DNA in a series of 80 breast carcinomas.