Wesselmann Konrad M, Baronti Cécile, Nougairède Antoine, Thirion Laurence, de Lamballerie Xavier, Charrel Remi, Pezzi Laura
Unité des Virus Émergents, Aix-Marseille Univ, Università di Corsica, IRD 190, Inserm 1207, IRBA, Marseille, France.
Laboratoire Infections Virales Aiguës et Tropicales, AP-HM Hôpitaux Universitaires de Marseille, Marseille, France.
J Clin Microbiol. 2025 Jul 3:e0042025. doi: 10.1128/jcm.00420-25.
Mayaro virus (MAYV) is a mosquito-borne alphavirus that is widespread in the Amazon basin, where it co-circulates with the closely related chikungunya virus (CHIKV). Due to the limited surveillance and technical limitations of diagnostic assays (scarcity of commercial assays, serology cross-reactivity), the true burden of MAYV is uncertain. We designed a new RT-qPCR assay targeting the nsp1 gene for MAYV detection, which can be used in monoplex or duplex format. In the duplex format, the new MAYV assay is combined with a CHIKV assay and a second MAYV assay, both previously published. The lower limit of detection with a 95% positivity rate was determined to be <10 RNA copies/μL in monoplex and duplex formats for both MAYV and CHIKV. Monoplex and duplex assays proved to be linear within the tested range of approximately 10 to 10 RNA copies/μL and showed 100% specificity against a wide panel of arboviruses as well as several other pathogens in clinical samples. The testing of CHIKV-positive sera and MAYV-spiked plasma samples confirmed the suitability of the assays in a clinical setting. These assays offer a reliable tool for detection and differentiation of MAYV and CHIKV in endemic settings.IMPORTANCEMolecular diagnostic capabilities for detecting alphaviruses other than chikungunya virus (CHIKV) remain limited. Mayaro virus (MAYV), an emerging mosquito-borne alphavirus, co-circulates with CHIKV in the Americas, making clinical differentiation between the two viruses challenging. To address this, we developed a novel RT-qPCR assay specifically for the detection of MAYV, which can also be used in a duplex format to simultaneously detect and distinguish CHIKV. The assay was thoroughly evaluated in both monoplex and duplex formats and demonstrated high sensitivity and specificity. This new tool is particularly valuable for the detection of MAYV, especially in resource-limited settings, where its duplex format offers efficient and accurate differentiation of acute infections.
马亚罗病毒（MAYV）是一种由蚊子传播的甲病毒，在亚马逊流域广泛存在，在那里它与密切相关的基孔肯雅病毒（CHIKV）共同传播。由于监测有限以及诊断检测存在技术局限性（商业检测稀缺、血清学交叉反应），MAYV的真实负担尚不确定。我们设计了一种针对MAYV检测的靶向非结构蛋白1（nsp1）基因的新型逆转录定量聚合酶链反应（RT-qPCR）检测方法，该方法可采用单重或双重检测形式。在双重检测形式中，新的MAYV检测方法与一种CHIKV检测方法以及另一种先前已发表的MAYV检测方法相结合。对于MAYV和CHIKV，单重和双重检测形式下95%阳性率时的检测下限均确定为<10个RNA拷贝/微升。单重和双重检测方法在约10至10个RNA拷贝/微升的测试范围内被证明具有线性，并且对临床样本中的多种虫媒病毒以及其他几种病原体显示出100%的特异性。对CHIKV阳性血清和MAYV加标血浆样本的检测证实了这些检测方法在临床环境中的适用性。这些检测方法为在流行地区检测和区分MAYV和CHIKV提供了可靠工具。重要性检测除基孔肯雅病毒（CHIKV）之外的甲病毒的分子诊断能力仍然有限。马亚罗病毒（MAYV）是一种新出现的由蚊子传播的甲病毒，在美洲与CHIKV共同传播，这使得两种病毒的临床鉴别具有挑战性。为解决这一问题，我们开发了一种专门用于检测MAYV的新型RT-qPCR检测方法，该方法也可采用双重检测形式同时检测和区分CHIKV。该检测方法在单重和双重检测形式下均经过了全面评估，并显示出高灵敏度和特异性。这种新工具对于检测MAYV特别有价值，尤其是在资源有限的环境中，其双重检测形式能够有效且准确地区分急性感染。
