Burroni Elena, Sani Cristina, Bisanzi Simonetta, Ocello Cristina
Laboratorio regionale HPV e biologia molecolare, Istituto per lo studio e la prevenzione oncologica (ISPO), Firenze.
Consulente statistico.
Epidemiol Prev. 2016 Mar-Apr;40(3-4):164-70. doi: 10.19191/EP16.3-4.AD03.077.
to evaluate the reproducibility of the High-Risk Hybrid Capture 2 (HC2-HR, Qiagen) test within the frame of cervical cancer screening with HPV, for samples with values very close to the cut-off (borderline), that is 0.80≤RLU/CO≤0.99 (RLU/CO: relative light units/cut-off) and to assess any possible presence of high grade lesions.
all borderline samples collected from January 2014 to August 2015 were repeated with HR-HC2 test within two weeks from the first test. For HPV-positive samples, cytology and colposcopy results (if present) were analysed.
national and international directions recommend the use of HPV testing as a primary test for cervical cancer screening for women aged 30/34-64 years. In Tuscany Region (Central Italy) the primary screening with HPV test was introduced in 2013 and the HPV test currently used, HR-HC2, has a positive cut-off≥1 RLU/CO.
proportion of HPV-positive tests at repetition, then stratified by type of material (ThinPrep, STM) and for the following subclasses of RLU/CO: 0.80-0.89 and 0.90-0.99.
253 samples were borderline at first test with HR-HC2 (about 0.4% of all samples tested). About 83% (209/253) of the samples was confirmed to be negative at the HPV test repetition: 88% for RLU/CO=0.80-0.89 and 76% for RLU/CO=0.90-0.99 (p-value=0.014). Median RLU/CO value of 44 HPV-positive samples at repetition is 1.4, with a maximum RLU/CO value of 16.7. In addition, 90% of HPV-positive samples at repetition has a RLU/CO values below or equal to 3.6. Even the used sampling system affects reproducibility: 26.4% (33/125) of the samples resulted positive at the repetition were in ThinPrep vs. 8.6% (11/128) of those in STM (p-value<0.001). The cytology result of HPV positive samples at repetition showed 84% (37/44) of normal samples and 7 low-grade lesions. No high-grade lesion was found in people who performed a colposcopy.
as expected, the reproducibility decreases for values closest to the cut-off (0.90-0.99), confirming the intrinsic limit to all analytical methods near to the cut-off. Moreover, even if the difference in reproducibility between the two systems of sampling is statistically significant, in samples that resulted positive at repetition high-grade lesions were not found at the time of HPV screening test.
在人乳头瘤病毒（HPV）宫颈癌筛查框架内，评估高危型杂交捕获2（HC2-HR，Qiagen公司）检测对于RLU/CO值非常接近临界值（即0.80≤RLU/CO≤0.99，RLU/CO：相对光单位/临界值）样本的重复性，并评估是否存在任何高级别病变。
收集2014年1月至2015年8月期间所有临界值样本，在首次检测后的两周内用HR-HC2检测进行重复检测。对于HPV阳性样本，分析其细胞学和阴道镜检查结果（如有）。
国家和国际指南建议将HPV检测作为30/34-64岁女性宫颈癌筛查的主要检测方法。在托斯卡纳地区（意大利中部），2013年引入了HPV检测作为主要筛查方法，目前使用的HPV检测方法HR-HC2的阳性临界值≥1 RLU/CO。
重复检测时HPV阳性检测的比例，然后按样本类型（薄层液基细胞学检测、SurePath液基细胞学检测）以及RLU/CO的以下亚类进行分层：0.80-0.89和0.90-0.99。
首次检测时，253个样本用HR-HC2检测为临界值（约占所有检测样本的0.4%）。约83%（209/253）的样本在HPV检测重复时被确认为阴性：RLU/CO=0.80-0.89时为88%，RLU/CO=0.90-0.99时为76%（p值=0.014）。重复检测时44个HPV阳性样本的RLU/CO中位数为1.4，最大RLU/CO值为16.7。此外，重复检测时90%的HPV阳性样本的RLU/CO值低于或等于3.6。所用的采样系统也会影响重复性：重复检测时呈阳性的样本中，薄层液基细胞学检测样本占26.4%（33/125），而SurePath液基细胞学检测样本占8.6%（11/128）（p值<0.001）。重复检测时HPV阳性样本的细胞学结果显示，84%（37/44）为正常样本，7个为低级别病变。进行阴道镜检查的人群中未发现高级别病变。
正如预期的那样，对于最接近临界值（0.90-0.99）的值，重复性降低，这证实了所有接近临界值的分析方法都存在内在局限性。此外，即使两种采样系统在重复性上的差异具有统计学意义，但在HPV筛查检测时，重复检测呈阳性的样本中未发现高级别病变。
