DDL Diagnostic Laboratory, Fonteynenburghlaan 7, 2275 CX Voorburg, The Netherlands.
J Clin Virol. 2009 Nov;46 Suppl 3:S21-6. doi: 10.1016/S1386-6532(09)70297-5.
Epidemiologic studies have classified 18 genotypes of the human papillomavirus (HPV) as (probably) high-risk (HR) based on their association with cervical cancer, i.e., HPV 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, and 82. Given the fact that certain HR HPV types confer an increased risk of cervical (pre)cancer, type-specific identification might aid clinical management of women tested positive for HR HPV. Therefore, the development of robust, high-throughput genotyping assays is important.
An analytical comparison of the digene HPV Genotyping LQ Test (digene LQ Test), capable of identifying 18 HR types using bead-based xMAP suspension array technology, with the established Reverse Line Blot (RLB) genotyping assay was carried out on amplimers generated with the clinically validated GP5+/6+-PCR method.
GP5+/6+ amplimers, generated from 434 digene High Risk HPV HC2 DNA Test (HC2)-positive and 95 HC2-negative cervical smears, were genotyped by both the digene LQ Test and the RLB genotyping assay.
The genotyping assays revealed high agreement for overall HR HPV detection (ú = 0.884) and type-specific identification of the 18 HR HPV types (overall ú = 0.958, individual ú range 0.795 to 1.000). The digene LQ Test demonstrated a very good inter-laboratory reproducibility (ú = 0.987). Among the HC2-positive women, the digene LQ Test revealed positivity for one or more HR HPV type(s) in 85.9%, and negativity was observed in 97.9% of the HC2-negative women.
The digene LQ Test demonstrated a high genotyping agreement with the established RLB genotyping assay on GP5+/6+ amplimers. This novel assay allows for high-throughput genotyping following HR HPV testing by HC2.
流行病学研究根据与宫颈癌的相关性，将 18 种人乳头瘤病毒（HPV）基因型分类为（可能）高危（HR）型，即 HPV16、18、26、31、33、35、39、45、51、52、53、56、58、59、66、68、73 和 82。鉴于某些 HR HPV 类型会增加宫颈癌（前）癌的风险，特定类型的鉴定可能有助于对 HR HPV 检测呈阳性的女性进行临床管理。因此，开发稳健、高通量的基因分型检测方法非常重要。
采用基于珠基 xMAP 悬浮阵列技术的 digene HPV 基因分型 LQ 检测（digene LQ 检测）对临床验证的 GP5+/6+-PCR 方法生成的扩增子进行分析比较，该方法能够鉴定 18 种 HR 类型，与已建立的反向线印迹（RLB）基因分型检测方法进行比较。
采用 GP5+/6+-PCR 方法对 434 例 digene High Risk HPV HC2 DNA Test（HC2）阳性和 95 例 HC2 阴性宫颈涂片进行扩增，用 digene LQ 检测和 RLB 基因分型检测对扩增产物进行基因分型。
基因分型检测结果显示，HR HPV 总体检测的一致性很高（u = 0.884），18 种 HR HPV 类型的特异性识别也很高（总体 u = 0.958，个体 u 值范围为 0.795 至 1.000）。digene LQ 检测具有很好的实验室间可重复性（u = 0.987）。在 HC2 阳性的女性中，digene LQ 检测显示一种或多种 HR HPV 型别阳性率为 85.9%，HC2 阴性的女性中均为阴性。
digene LQ 检测在 GP5+/6+扩增子上与已建立的 RLB 基因分型检测具有很高的基因分型一致性。该新型检测方法可在 HR HPV 检测后通过 HC2 进行高通量基因分型。
