Gynaecology Clinic, Charité Universitätsmedizin, Corporate Member of Freie Universität Berlin, Humboldt-Universität Berlin and Berlin Institute of Health, Berlin, Germany.
AID/GenID Diagnostika, Strassberg, Germany.
Intervirology. 2019;62(3-4):124-133. doi: 10.1159/000502207. Epub 2019 Sep 5.
Cervical cancer is caused by a persistent infection of human papillomavirus (HPV). Therefore, tests which detect the carcinogenic virus can be used for cervical cancer screening.
This is the first evaluation of the HPV DNA Array (AID Diagnostika, Strassberg, Germany), an E1-based genotyping polymerase chain reaction (PCR) test for identification of 29 HPV types (6, 11, 16, 18, 26, 31, 33, 35, 39, 40, 42, 44, 45, 51, 52, 53, 54, 56, 58, 59, 66, 67, 68, 69, 70, 73, 82, 85, and 97).
Analytical performance of the assay was assessed with cervical cancer cell lines with known HPV status, and preselected clinical cervical scrapings genotyped by multiplexed genotyping (MPG) with a Luminex readout (validated in-house assay). Intra- and inter-laboratory reproducibility experiments were performed to ensure the reliability of the assay.
HPV DNA Array identified the intrinsic HPV genotype in all cervical cancer cell lines and demonstrated a high sensitivity for HPV16 probe (1 cell per PCR reaction), as well as HPV18 and 45 probes (100 cells per PCR reaction). When compared with MPG, HPV DNA Array showed a good agreement of 92.2% for HPV detection irrespective of type (κ = 0.601), and demonstrated high agreement for HPV16 (80.7%, κ = 0.836) and HPV18 (86.7%, κ = 0.925). Furthermore, high intra-/inter-laboratory reproducibility was observed (90.9-100%).
HPV DNA Array showed high sensitivity for correct HPV genotype detection in experimental and clinical samples with a good correlation to the reference test. Since HPV DNA Array is based on a simple multiplexed PCR followed by reverse hybridization in a 96-well format and automated visual readout by AID ELISpot reader, it is capable of high throughput in a time-effective manner. HPV DNA Array could be considered for extended HPV genotyping of cervical smears.
宫颈癌是由人乳头瘤病毒（HPV）持续感染引起的。因此，检测致癌病毒的检测方法可用于宫颈癌筛查。
本研究是对 HPV DNA 微阵列（AID Diagnostika，Strassberg，德国）的首次评估，这是一种基于 E1 的基因分型聚合酶链反应（PCR）检测方法，可鉴定 29 种 HPV 类型（6、11、16、18、26、31、33、35、39、40、42、44、45、51、52、53、54、56、58、59、66、67、68、69、70、73、82、85 和 97）。
采用已知 HPV 状态的宫颈癌细胞系评估该检测方法的分析性能，并采用多重基因分型（MPG）与 Luminex 读取（内部验证检测方法）对预选的临床宫颈刮片进行基因分型。进行了室内和实验室间重复性实验，以确保检测方法的可靠性。
HPV DNA 微阵列可鉴定所有宫颈癌细胞系中的 HPV 固有基因型，并对 HPV16 探针（每个 PCR 反应 1 个细胞）以及 HPV18 和 45 探针（每个 PCR 反应 100 个细胞）具有高灵敏度。与 MPG 相比，HPV DNA 微阵列对 HPV 检测具有良好的一致性（不论 HPV 类型），为 92.2%（κ=0.601），并且 HPV16（80.7%，κ=0.836）和 HPV18（86.7%，κ=0.925）的一致性也很高。此外，观察到高度的室内/实验室间重复性（90.9%-100%）。
HPV DNA 微阵列在实验和临床样本中对正确 HPV 基因型的检测具有较高的敏感性，与参考检测方法相关性良好。由于 HPV DNA 微阵列基于简单的多重 PCR，随后在 96 孔格式中进行反向杂交，并由 AID ELISpot 读取器进行自动目视读取，因此能够以有效的方式实现高通量。HPV DNA 微阵列可考虑用于宫颈涂片的 HPV 基因分型扩展。
