Clinical Virology, Laboratory Medicine, University Hospital Basel, Basel, Switzerland.
Department of Biomedicine, Transplantation & Clinical Virology, University of Basel, Basel, Switzerland.
J Med Virol. 2023 Feb;95(2):e28583. doi: 10.1002/jmv.28583.
Human papillomavirus (HPV) infections are often asymptomatic, but some of the >200 HPV genotypes confer a high risk for precancerous cervical lesions and cervical cancer. Current clinical management of HPV infections relies on reliable nucleic acid testing detection and genotyping. We prospectively compared nucleic acid extraction without and with prior centrifugation enrichment for detecting and genotyping HPV in cervical swabs with atypical squamous or glandular cells. Consecutive swabs were analyzed from 45 patients with atypical squamous or glandular cells. Nucleic acids were extracted in parallel using three procedures, Abbott-M2000, Roche-MagNA-Pure-96 Large-Volume Kit without (Roche-MP-large) and with prior centrifugation (Roche-MP-large/spin) and tested using Seegene-Anyplex-II HPV28. In total, 54 HPV-genotypes were detected in 45 samples, 51 by Roche-MP-large/spin, 48 by Abbott-M2000 and 42 by Roche-MP-large. The overall concordance was 80% for detecting any HPV and 74% for specific HPV-genotypes. Roche-MP-large/spin and Abbott-M2000 showed the highest concordance for HPV detection (88.9%; kappa 0.78), and genotyping (88.5%). Two and more HPV-genotypes were detected in 15 samples, often with one HPV being more abundant. Dilution series confirmed the specific detection of multiple HPV-genotypes and their relative abundance. In 285 consecutive follow-up samples extracted by Roche-MP-large/spin, the top three detected genotypes were the high-risk HPV16, HPV53, HPV56 and the low-risk HPV42, HPV54 and HPV61. Rate and breadth of HPV detection in cervical swabs depends on extraction protocols being highest after centrifugation/enrichment. As multivalent HPV-vaccine coverage increases, detecting the evolving HPV virome depends on improved extraction and broader genotype coverage.
人乳头瘤病毒 (HPV) 感染通常无症状，但 200 多种 HPV 基因型中的一些会导致宫颈癌前病变和宫颈癌的高风险。目前 HPV 感染的临床管理依赖于可靠的核酸检测和基因分型。我们前瞻性地比较了未经离心富集和离心富集后提取核酸检测和 HPV 基因分型的方法，比较对象为宫颈拭子中具有非典型鳞状或腺细胞的患者。连续分析了 45 例具有非典型鳞状或腺细胞的患者的宫颈拭子。使用三种方法平行提取核酸，分别为 Abbott-M2000、Roche-MagNA-Pure-96 Large-Volume Kit 未经离心富集（Roche-MP-large）和离心富集（Roche-MP-large/spin），并使用 Seegene-Anyplex-II HPV28 进行检测。在总共 45 个样本中检测到 54 种 HPV 基因型，其中 51 种通过 Roche-MP-large/spin 检测到，48 种通过 Abbott-M2000 检测到，42 种通过 Roche-MP-large 检测到。检测任何 HPV 的总体一致性为 80%，特定 HPV 基因型的一致性为 74%。Roche-MP-large/spin 和 Abbott-M2000 对 HPV 检测（88.9%；kappa 值 0.78）和基因分型（88.5%）的一致性最高。15 个样本中检测到两种或两种以上 HPV 基因型，通常一种 HPV 更为丰富。稀释系列证实了多种 HPV 基因型的特异性检测及其相对丰度。在通过 Roche-MP-large/spin 提取的 285 个连续随访样本中，检测到的三种最常见的基因型是高危 HPV16、HPV53、HPV56 和低危 HPV42、HPV54 和 HPV61。宫颈拭子中 HPV 的检测率和广度取决于提取方案，离心富集后最高。随着多价 HPV 疫苗覆盖率的提高，检测不断进化的 HPV 病毒组取决于改进的提取和更广泛的基因型覆盖。
