Karaaslan Elif, Chiang Cheng-Feng, Kurutaş Gülter Öncü, Barkay Orçun, Çetin Güler Nesibe Selma, Kalkan Merve Yazıcı, Karakoç Parlayan Hanife Nur, Akdoğan Özlem, Çelikbaş Aysel Kocagül, Aksoy Firdevs, Binay Umut Devrim, Baykam Nurcan, Yılmaz Gürdal, Sajadi Mohammad M, Pegan Scott D, Klena John D, Montgomery Joel M, Karakeçili Faruk, Kalkan Ahmet, Doymaz Mehmet Ziya, Spiropoulou Christina F, Bergeron Éric
Viral Special Pathogens Branch, Division of High-Consequence Pathogens and Pathology, Centers for Disease Control and Prevention, Atlanta, GA, USA; Division of Biomedical Sciences, University of California Riverside, Riverside, CA, USA.
Viral Special Pathogens Branch, Division of High-Consequence Pathogens and Pathology, Centers for Disease Control and Prevention, Atlanta, GA, USA.
EBioMedicine. 2025 Jul 23;118:105857. doi: 10.1016/j.ebiom.2025.105857.
Crimean-Congo hemorrhagic fever virus (CCHFV), a zoonotic agent in the Nairoviridae family (genus Orthonairovirus), is a high-priority pathogen. CCHFV infection causes Crimean-Congo hemorrhagic fever (CCHF), a human disease with case fatality rates of up to 40%. Serological surveillance of CCHFV in animals and humans is crucial for ecological studies and public health.
We developed CCHFV mix-and-read assays utilizing split-NanoLuc technology (NanoBiT) to detect anti-CCHFV antibodies against the nucleoprotein (NP) stalk region and the GP38 glycoprotein. These species- and isotype-agnostic assays provide results in ∼30 min. Using serum samples from RT-PCR-confirmed CCHF cases collected during and after hospitalization, we investigated anti-NP and anti-GP38 antibody development. The performance of the mix-and-read assays was compared to the NP-based IDScreen® CCHF commercial assay using human sera, and cross-reactivity potential was evaluated using a diverse panel of anti-orthonairovirus antisera raised in mice.
In human convalescent cases (n = 21), mix-and-read assay concordance between anti-GP38 and anti-NP antibody detection was 100%. Both mix-and-read assays and IDScreen® CCHF demonstrated identical sensitivity of 95.2% in convalescent patients. The specificity of the NP assay was 98.9%, and that of GP38 was 99.7%, both comparable to IDScreen® CCHF (specificity: 99.7%). Cross-reactivity against CCHF NP and GP38, regardless of assay type, was primarily observed in antisera raised against other orthonairoviruses within the Nairobi sheep disease genogroup.
The simplicity and robust performance of the CCHFV mix-and-read assays make them ideal tools for supporting serological surveillance in humans and animals. Furthermore, the inclusion of the GP38 antigen alongside NP enhances the precise identification of retrospective CCHF cases, further strengthening broad surveillance efforts.
CDC Emerging Infectious Disease Research Core Funds, funding for reagent, CDC personal, travel. Defence Threat Reduction Agency (HDTRA12210007): E.K. salary. Oak Ridge Institute for Science and Education (ORISE): E.K. salary and travel. National Institute of Allergy and Infectious Diseases (1R01AI180125-01A1): sample acquisition. Funding sources did not have a role in the writing or decision to submit the publication.
克里米亚-刚果出血热病毒（CCHFV）是内罗毕病毒科（正内罗毕病毒属）的一种人畜共患病原体，是一种高优先级病原体。CCHFV感染会引发克里米亚-刚果出血热（CCHF），这是一种人类疾病，病死率高达40%。对动物和人类进行CCHFV血清学监测对于生态学研究和公共卫生至关重要。
我们利用分裂纳米荧光素酶技术（NanoBiT）开发了CCHFV混合读取检测方法，以检测针对核蛋白（NP）茎区和GP38糖蛋白的抗CCHFV抗体。这些不依赖物种和亚型的检测方法在约30分钟内即可得出结果。我们使用住院期间及出院后收集的经逆转录聚合酶链反应（RT-PCR）确诊的CCHF病例的血清样本，研究抗NP和抗GP38抗体的产生情况。将混合读取检测方法的性能与基于NP的IDScreen® CCHF商业检测方法在人血清中的性能进行比较，并使用在小鼠中产生的多种抗正内罗毕病毒抗血清评估交叉反应潜力。
在人类康复病例（n = 21）中，抗GP38和抗NP抗体检测的混合读取检测方法一致性为100%。混合读取检测方法和IDScreen® CCHF在康复患者中的敏感性均为95.2%。NP检测方法的特异性为98.9%，GP38检测方法的特异性为99.7%，两者均与IDScreen® CCHF相当（特异性：99.7%）。无论检测类型如何，针对CCHF NP和GP38的交叉反应主要在针对内罗毕羊病基因组内其他正内罗毕病毒产生的抗血清中观察到。
CCHFV混合读取检测方法的简便性和强大性能使其成为支持人类和动物血清学监测的理想工具。此外，将GP38抗原与NP一起使用可提高对既往CCHF病例的精确识别，进一步加强广泛的监测工作。
美国疾病控制与预防中心新兴传染病研究核心基金，用于试剂、疾病控制与预防中心人员及差旅的资金。国防威胁降低局（HDTRA12210007）：E.K.的薪资。橡树岭科学与教育研究所（ORISE）：E.K.的薪资及差旅费用。美国国立过敏与传染病研究所（1R01AI180125 - 01A1）：样本采集。资金来源在本文撰写或提交发表的决策过程中未发挥作用。
