Molecular Pathology Laboratory, Department of Pathology, Hvidovre Hospital, Copenhagen University Hospital, Kettegård Allé 30, 2650, Hvidovre, Denmark.
Department of Laboratory Medicine, Karolinska Institutet, and Karolinska University Laboratory, Karolinska University Hospital, Forskningsgatan, F56 14186, Stockholm, Sweden.
BMC Cancer. 2020 May 6;20(1):396. doi: 10.1186/s12885-020-06888-0.
To ensure the highest quality of human papillomavirus (HPV) testing in primary cervical cancer screening, novel HPV assays must be evaluated in accordance with the international guidelines. Furthermore, HPV assay with genotyping capabilities are becoming increasingly important in triage of HPV positive women in primary HPV screening. Here we evaluate a full genotyping HPV assay intended for primary screening.
The CLART® HPV4S (CLART4S) assay is a newly developed full-genotyping assay detecting 14 oncogenic (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68) and two non-oncogenic HPV genotypes (6, 11). It was evaluated using SurePath and ThinPrep screening samples collected from the Danish and Swedish cervical cancer screening programs, respectively. For calculation of sensitivity, 81 SurePath and 80 ThinPrep samples with confirmed ≥CIN2 were assessed. For clinical specificity analysis, 1184 SurePath and 1169 ThinPrep samples from women with <CIN2 histology were assessed. Sensitivity and specificity of the CLART4S assay was compared to an established reference test; the MGP-PCR (Modified General Primers GP5+/6+ with genotyping using Luminex). Inter and intra laboratory reproducibility of the assay was assessed using 540 SurePath and 520 ThinPrep samples, respectively. The genotype concordance between CLART4S and MGP-PCR was also assessed.
In SurePath samples, the sensitivity of CLART4S was 0.90 (MGP-PCR =0.93) and the specificity was 0.91 (MGP-PCR = 0.91); In ThinPrep samples the sensitivity of CLART4S was 0.98 (MGP-PCR = 1.00) and specificity was 0.94 (MGP-PCR =0.87). The CLART4S was shown to be non-inferior to that of MGP-PCR for both sensitivity (p = 0.002; p = 0.01) and specificity (p = 0.01; p = 0.00) in SurePath and ThinPrep samples, respectively. Intra-laboratory reproducibility and inter-laboratory agreement was met for both media types. The individual genotype concordance between CLART4S and MGP-PCR was good agreement for almost all 14 HPV genotypes in both media types.
The CLART4S assay was proved non-inferior to the comparator assay MGP-PCR for both sensitivity and specificity using SurePath and ThinPrep cervical cancer screening samples from the Danish and Swedish screening programs, respectively. This is the first study to demonstrate clinical validation of a full-genotyping HPV assay conducted in parallel on both SurePath and ThinPrep collected samples.
为了确保人乳头瘤病毒（HPV）检测在宫颈癌筛查中的最高质量，新的 HPV 检测方法必须按照国际准则进行评估。此外，在 HPV 阳性妇女的初级 HPV 筛查中，具有基因分型能力的 HPV 检测方法在分流中变得越来越重要。在此，我们评估了一种用于初级筛查的全基因分型 HPV 检测方法。
CLART®HPV4S（CLART4S）检测是一种新开发的全基因分型检测方法，可检测 14 种致癌型（16、18、31、33、35、39、45、51、52、56、58、59、66、68）和两种非致癌型 HPV 基因型（6、11）。该检测方法使用丹麦和瑞典宫颈癌筛查计划分别采集的 SurePath 和 ThinPrep 筛查样本进行了评估。为了计算敏感性，对 81 例经 SurePath 和 80 例经 ThinPrep 检测证实≥CIN2 的样本进行了评估。为了进行临床特异性分析，对 1184 例来自患有<CIN2 组织学的女性的 SurePath 和 1169 例经 ThinPrep 检测的样本进行了评估。CLART4S 检测的敏感性和特异性与既定的参考检测方法（改良通用引物 GP5+/6+ 与使用 Luminex 的基因分型）进行了比较。使用分别来自 540 例 SurePath 和 520 例 ThinPrep 样本评估了检测方法的实验室内和实验室间重复性。还评估了 CLART4S 和 MGP-PCR 之间的基因型一致性。
在 SurePath 样本中，CLART4S 的敏感性为 0.90（MGP-PCR=0.93），特异性为 0.91（MGP-PCR=0.91）；在 ThinPrep 样本中，CLART4S 的敏感性为 0.98（MGP-PCR=1.00），特异性为 0.94（MGP-PCR=0.87）。CLART4S 对于敏感性（p=0.002;p=0.01）和特异性（p=0.01;p=0.00），分别与 MGP-PCR 相比，在 SurePath 和 ThinPrep 样本中均显示出非劣效性。两种介质的实验室内重复性和实验室间一致性均得到满足。在两种介质中，CLART4S 和 MGP-PCR 之间的个体基因型一致性对于几乎所有 14 种 HPV 基因型均具有良好的一致性。
CLART4S 检测方法在丹麦和瑞典筛查计划的 SurePath 和 ThinPrep 宫颈癌筛查样本中，分别与比较检测方法 MGP-PCR 相比，在敏感性和特异性方面均显示出非劣效性。这是第一项在 SurePath 和 ThinPrep 采集样本上同时进行全基因分型 HPV 检测的临床验证研究。
