d'Avenia M, Dell'Anno F, Martinelli M, Santomauro L, Njoku R C, Arroyo Mühr L S, Iacobellis M, Cocuzza C E
School of Medicine and Surgery, University of Milano-Bicocca, Milan, Italy.
UOSVD Cytopathology and Screening Laboratory, ASL BARI, Bari, Italy.
J Med Virol. 2025 Jul;97(7):e70482. doi: 10.1002/jmv.70482.
In HPV-primary screening, sample quality significantly influences test accuracy. Unlike cytology-based screening, no consensus guidelines presently exist for sample quality assessment in HPV testing. This study aims to evaluate the impact of sample cellularity on HPV testing. A total of 37 592 liquid-based cytology (LBC) samples from women undergoing HPV-primary screening (aged 30-64, median 48; IQR: 40-56 years) were analyzed using Cobas 4800 HPV Test (Roche). Sample adequacy was assessed by the assay's β-globin internal control and by an independent quantitative cellularity assessment (OncoPredict HPV, Hiantis). HPV positivity rates (PR) were stratified according to β-globin Ct values. Among the analyzed samples, 50.0%, 47.1%, 2.3%, and 0.6% had β-globin Ct values of ≤ 28, > 28 to ≤ 32, > 32 to ≤ 34, and > 34, respectively. Overall HPV-PR was 7.7% (2891/37 592). PR reached 9.7% in samples with β-globin ≤ 28 Ct (1820/18 801), decreasing markedly to 1.4% for β-globin > 34 Ct (3/214), (p < 0.001). Quantitative analysis showed that Cobas 4800 β-globin Ct = 34 corresponds to approximately 1.5 × 10^3 nucleated cells/reaction. A subset of 195 HPV-negative samples with β-globin Ct ≥ 34 was evaluated by liquid based cytology (LBC): 19% had inadequate cellularity according to LBC guidelines, 8% were ≥ ASC-US and 73% NILMs. 65% of adequate LBC showed cellular atrophy. These findings emphasize the importance of assessing cellularity in HPV-screening to avoid potentially false-negative results due to inadequate samples. Future research should focus on establishing standardized cellularity thresholds to improve screening accuracy.
在人乳头瘤病毒（HPV）初筛中，样本质量会显著影响检测准确性。与基于细胞学的筛查不同，目前HPV检测中尚无关于样本质量评估的共识指南。本研究旨在评估样本细胞含量对HPV检测的影响。使用Cobas 4800 HPV检测法（罗氏公司）分析了37592例接受HPV初筛女性（年龄30 - 64岁，中位数48岁；四分位间距：40 - 56岁）的液基细胞学（LBC）样本。通过检测的β - 珠蛋白内对照和独立的定量细胞含量评估（OncoPredict HPV，Hiantis公司）来评估样本的充足性。HPV阳性率（PR）根据β - 珠蛋白Ct值进行分层。在分析的样本中，β - 珠蛋白Ct值≤28、>28至≤32、>32至≤34和>34的样本分别占50.0%、47.1%、2.3%和0.6%。总体HPV - PR为7.7%（2891/37592）。β - 珠蛋白Ct≤28的样本中PR达到9.7%（1820/18801），而β - 珠蛋白Ct>34的样本中PR显著降至1.4%（3/214），（p<0.001）。定量分析表明，Cobas 4800 β - 珠蛋白Ct = 34大约对应1.5×10³有核细胞/反应。对195例β - 珠蛋白Ct≥34的HPV阴性样本的一个子集进行了液基细胞学（LBC）评估：根据LBC指南，19%的样本细胞含量不足，8%为≥非典型鳞状细胞不能明确意义（ASC - US），73%为未见上皮内病变或恶性病变（NILM）。65%细胞含量充足的LBC样本显示细胞萎缩。这些发现强调了在HPV筛查中评估细胞含量的重要性，以避免因样本不足导致潜在的假阴性结果。未来的研究应专注于建立标准化的细胞含量阈值，以提高筛查准确性。
