Atceken Nazente, Asghari Dilmani Sara, Abdullah Ahmed Choukri, Sarıkaya Mutlu, Yigci Defne, Korkmaz Gozde, Tasoglu Savas
School of Biomedical Sciences and Engineering, Koç University, 34450 Istanbul, Turkey.
Koç University Translational Medicine Research Center (KUTTAM), Koç University, 34450 Istanbul, Turkey.
Biosensors (Basel). 2025 Jan 6;15(1):23. doi: 10.3390/bios15010023.
Human monkeypox (Mpox) is a zoonotic disease caused by the Monkeypox virus (MPXV). As of 14 August 2024, the World Health Organization (WHO) has declared it a global health emergency. For Mpox, this was the second public health emergency of global significance in the past two years. MPXV belongs to the family and is phylogenetically and epidemically divided into two clades: the Congo Basin (Clade-I) and the West African (Clade-II) clades. Clade-I has been associated with more severe disease progression and higher mortality compared to Clade-II, and thus the differentiation between clades can play an important role in predicting disease prognosis. The LAMP technique has the advantages of not requiring thermal cycling and achieving higher amplification in a shorter time compared to qPCR. Different types of LAMP assays were developed in this study to benefit from these advantages. We report the development of LAMP-1 and LAMP-2 assays using the LAMP method to detect MPXV Clade-I and Clade-II, respectively. The LAMP-1 assay includes both fluorescence and visible colorimetric readout tests developed with sensitivities of 10 and 10 copies, respectively. For the LAMP-2 assay, a probe-based test utilizing the Novel R-Duplex DARQ probe was developed, offering fluorescence detection at a sensitivity of 10 copies. As a result, we successfully developed three highly specific molecular diagnostic tests that distinctly differentiate between MPXV clades, delivering essential tools for the precise diagnosis and effective control of Mpox.
人类猴痘是由猴痘病毒（MPXV）引起的一种人畜共患病。截至2024年8月14日，世界卫生组织（WHO）已宣布其为全球卫生紧急事件。对于猴痘而言，这是过去两年内第二次具有全球意义的公共卫生紧急事件。MPXV属于痘病毒科，在系统发育和流行病学上分为两个进化枝：刚果盆地进化枝（进化枝I）和西非进化枝（进化枝II）。与进化枝II相比，进化枝I与更严重的疾病进展和更高的死亡率相关，因此进化枝之间的区分在预测疾病预后方面可发挥重要作用。环介导等温扩增（LAMP）技术具有无需热循环且与实时荧光定量聚合酶链反应（qPCR）相比能在更短时间内实现更高扩增的优点。本研究开发了不同类型的LAMP检测方法以利用这些优势。我们报告了分别使用LAMP方法检测MPXV进化枝I和进化枝II的LAMP-1和LAMP-2检测方法的开发情况。LAMP-1检测方法包括分别以10拷贝和10拷贝的灵敏度开发的荧光和可见比色读数测试。对于LAMP-2检测方法，开发了一种利用新型R-双链DARQ探针的基于探针的测试，其荧光检测灵敏度为10拷贝。结果，我们成功开发了三种高度特异性的分子诊断测试，可明确区分MPXV进化枝，为猴痘的精确诊断和有效控制提供了重要工具。
