Desai Kanan T, Ajenifuja Kayode O, Adepiti Clement A, Inturrisi Federica, Dagnall Casey, Hoffman Amanda C, Egemen Didem, Gage Julia C, Wentzensen Nicolas, de Sanjose Silvia, Schiffman Mark
National Cancer Institute, Rockville, Maryland, USA.
Norwalk Hospital Internal Medicine Residency Program, Norwalk, Connecticut, USA.
J Clin Microbiol. 2025 Feb 19;63(2):e0163924. doi: 10.1128/jcm.01639-24. Epub 2024 Dec 31.
Human papillomavirus (HPV) genotype predicts cervical cancer risk, and genotyping could help guide the management of HPV positives as part of cervical screening. An isothermal amplification HPV extended genotyping test (ScreenFire HPV RS assay) can assay up to 96 samples/controls in 1 hour plus preparation time. A novel format with pre-aliquoted reagents and an anti-contamination component (Zebra BioDome) could simplify the HPV testing process and reduce the chances of post-amplification contamination. We validated the Zebra BioDome formulation prior to its clinical use. Residual provider-collected cervical samples ( = 450) from a population-based study in rural Nigeria were retested with ScreenFire, once using the standard assay version (liquid reagents combined onsite) and twice with Zebra BioDome. HPV results with adequate DNA ( = 427) were analyzed channel-by-channel and using the cervical cancer risk-based hierarchy of HPV type channels (HPV16, else 18/45, else 31/33/35/52/58, else 39/51/56/59/68, else high-risk HPV negative) to evaluate Zebra BioDome repeatability and accuracy against the standard version. Zebra BioDome reduced the number of pipetting steps to run the ScreenFire HPV assay. Following amplification, the BioDome material formed a sealant layer above the reaction components. Zebra BioDome had excellent repeatability and agreement with the standard version, both at the channel-specific analysis (positive percent agreement between 88.4% [HPV39/51/56/59/68] and 100% [HPV16]; negative percent agreement between 97.8% [HPV31/33/35/52/58] and 100% [HPV39/51/56/59/68]) and hierarchical analysis (overall agreement 97.2%). The assay version utilizing Zebra BioDome performed similarly to the previously validated standard version of the ScreenFire HPV assay and is now undergoing field evaluation. This solution has the potential to reduce assay preparation time and risk of contamination, providing a simpler, low-cost, near-point-of-care HPV testing and extended genotyping solution for cervical screening in lower-resource settings. The potential application of Zebra BioDome technology to other PCR assays should be considered.
This work validates a novel pre-packed formulation for the ScreenFire human papillomavirus (HPV) assay, which has the potential to simplify the HPV testing process and to reduce the chances of post-amplification contamination, providing a simpler, low-cost, near-point-of-care HPV testing, and extended genotyping solution for cervical screening in resource-limited settings as part of the ultimate public health goal to accelerate cervical cancer prevention. This technology can also have broad applications for other DNA amplification assays beyond HPV.
人乳头瘤病毒（HPV）基因型可预测宫颈癌风险，基因分型有助于指导作为宫颈筛查一部分的HPV阳性病例的管理。一种等温扩增HPV扩展基因分型检测（ScreenFire HPV RS检测法）在1小时加准备时间内可检测多达96个样本/对照。一种采用预分装试剂和防污染组件（Zebra BioDome）的新型形式可简化HPV检测过程，并减少扩增后污染的几率。我们在临床使用前对Zebra BioDome制剂进行了验证。来自尼日利亚农村一项基于人群研究的残留的由医疗机构收集的宫颈样本（n = 450），使用ScreenFire进行重新检测，一次使用标准检测版本（液体试剂在现场混合），两次使用Zebra BioDome。对具有足够DNA的HPV结果（n = 427）逐通道进行分析，并使用基于宫颈癌风险的HPV型别通道分级（HPV16，其他为18/45，其他为31/33/35/52/58，其他为39/51/56/59/68，其他为高危HPV阴性）来评估Zebra BioDome相对于标准版本的重复性和准确性。Zebra BioDome减少了运行ScreenFire HPV检测所需的移液步骤。扩增后，BioDome材料在反应成分上方形成密封层。Zebra BioDome在通道特异性分析（阳性百分比一致性在88.4%[HPV39/51/56/59/68]至100%[HPV16]之间；阴性百分比一致性在97.8%[HPV31/33/35/52/58]至100%[HPV39/51/56/59/68]之间）和分级分析（总体一致性为97.2%）方面与标准版本具有出色的重复性和一致性。使用Zebra BioDome的检测版本与之前验证的ScreenFire HPV检测标准版本表现相似，目前正在进行现场评估。该解决方案有可能减少检测准备时间和污染风险，为资源匮乏地区的宫颈筛查提供一种更简单、低成本、近床旁的HPV检测和扩展基因分型解决方案。应考虑Zebra BioDome技术在其他PCR检测中的潜在应用。
这项工作验证了一种用于ScreenFire人乳头瘤病毒（HPV）检测的新型预包装制剂，它有可能简化HPV检测过程并减少扩增后污染的几率，为资源有限地区的宫颈筛查提供一种更简单、低成本、近床旁的HPV检测和扩展基因分型解决方案，这是加速宫颈癌预防这一最终公共卫生目标的一部分。这项技术在HPV之外的其他DNA扩增检测中也可能有广泛应用。
