Miller Lesley S, Rao Anuradha, Tunnell Karlyn, Wang Yun F, Parsons Richard, McLendon Kaleb, Bassit Leda, Patel Mahi, Bowers Heather B, Sabino Courtney, Ramachandra Thanuja, Saeed Farzan, Schinazi Raymond F, Lam Wilbur, Sullivan Julie A, Fluker Shelly-Ann
Department of Medicine, Emory University School of Medicine, Atlanta, Georgia, United States of America.
Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, United States of America.
PLoS One. 2025 Jul 22;20(7):e0324088. doi: 10.1371/journal.pone.0324088. eCollection 2025.
Improvements in HCV testing, including Point of Care (POC) HCV RNA tests, are necessary to eliminate HCV. With this goal in mind, we established methods to collect capillary whole blood (CWB) via fingerstick, ensured its stability in a microtainer, and determined limit of detection (LoD) for various HCV genotypes. Next, we conducted a prospective study where CWB samples were collected from a cohort of 109 adult subjects at a safety-net hospital and tested for HCV RNA using the Xpert® HCV test on the GeneXpert® Xpress system and the cobas® HCV platform. We consistently obtained 250 μl CWB which was stable for up to 5 hours in the microtainer. Laboratory LoD studies demonstrated that the Xpert® HCV test could detect most HCV genotypes to <100 IU/ml in CWB. In the prospective clinical study, 89 subjects (82%) with valid Xpert® and cobas® HCV comparator results were analyzed. Using Xpert®, 16 out of 89 (18%) subjects had detectable HCV RNA and 73 (82%) had undetectable HCV RNA. Using cobas, 17 out of 89 (19%) participants had detectable HCV RNA, and 72 (81%) were undetectable. One sample was detectable on cobas but not Xpert®, yielding a sensitivity of 94% and specificity of 100%. This study demonstrates the feasibility of HCV RNA testing at POC using CWB obtained by fingerstick and provides preliminary data on the accuracy of the Xpert® HCV test performed by untrained operators in a CLIA-waived setting.
改进丙型肝炎病毒（HCV）检测方法，包括即时检测（POC）HCV RNA检测，对于消除HCV至关重要。出于这一目标，我们建立了通过手指采血收集毛细血管全血（CWB）的方法，确保其在微量采血管中的稳定性，并确定了各种HCV基因型的检测限（LoD）。接下来，我们进行了一项前瞻性研究，从一家安全网医院的109名成年受试者队列中采集CWB样本，并使用GeneXpert® Xpress系统上的Xpert® HCV检测和cobas® HCV平台检测HCV RNA。我们始终能获得250 μl CWB，其在微量采血管中可稳定长达5小时。实验室检测限研究表明，Xpert® HCV检测能够在CWB中检测到大多数HCV基因型，检测下限<100 IU/ml。在前瞻性临床研究中，对89名具有有效Xpert®和cobas® HCV比较结果的受试者进行了分析。使用Xpert®检测时，89名受试者中有16名（18%）可检测到HCV RNA，73名（82%）检测不到。使用cobas检测时，89名参与者中有17名（19%）可检测到HCV RNA，72名（81%）检测不到。有一个样本在cobas上可检测到，但在Xpert®上检测不到，灵敏度为94%，特异性为100%。本研究证明了使用手指采血获得的CWB进行即时检测HCV RNA的可行性，并提供了在CLIA豁免环境中未经培训的操作人员进行Xpert® HCV检测准确性的初步数据。
