Zika virus (ZIKV) is an emerging arbovirus belonging to the genus flavivirus that comprises other important public health viruses, such as dengue (DENV) and yellow fever (YFV). In general, ZIKV infection is a self-limiting disease, however cases of Guillain-Barré syndrome and congenital brain abnormalities in newborn infants have been reported. Diagnosing ZIKV infection remains a challenge, as viral RNA detection is only applicable until a few days after the onset of symptoms. After that, serological tests must be applied, and, as expected, high cross-reactivity between ZIKV and other flavivirus serology is observed. Plaque reduction neutralization test (PRNT) is indicated to confirm positive samples for being more specific, however it is laborious intensive and time consuming, representing a major bottleneck for patient diagnosis. To overcome this limitation, we developed a high-throughput image-based fluorescent neutralization test for ZIKV infection by serological detection. Using 226 human specimens, we showed that the new test presented higher throughput than traditional PRNT, maintaining the correlation between results. Furthermore, when tested with dengue virus samples, it showed 50.53% less cross reactivity than MAC-ELISA. This fluorescent neutralization test could be used for clinical diagnosis confirmation of ZIKV infection, as well as for vaccine clinical trials and seroprevalence studies.

Zika病毒（ZIKV）是黄病毒属的一种新兴虫媒病毒，包含其他重要的公共卫生病毒，例如登革热（DENV）和黄热病（YFV）。总的来说，ZIKV感染是一种自限性疾病，但是已经报道了格林-巴利综合征和新生儿先天性脑畸形的病例。诊断ZIKV感染仍然是一个挑战，因为病毒RNA检测仅适用于症状发作后的几天。之后，必须进行血清学检测，并且如预期的那样，观察到ZIKV与其他黄病毒血清学之间的高交叉反应性。噬菌斑减少中和测试（PRNT）可以确认阳性样本的特异性，但它费力且费时，是患者诊断的主要瓶颈。为克服此限制，我们开发了基于血清学检测的ZIKV感染的基于高通量图像的荧光中和测试。我们使用226个人体标本表明，新测试显示出比传统PRNT高的通量，并保持了结果之间的相关性。此外，当用登革热病毒样品进行测试时，它的交叉反应性比MAC-ELISA低50.53％。该荧光中和试验可用于ZIKV感染的临床诊断确认，以及疫苗临床试验和血清阳性率研究。保持结果之间的相关性。此外，当用登革热病毒样品进行测试时，它的交叉反应性比MAC-ELISA低50.53％。该荧光中和试验可用于ZIKV感染的临床诊断确认，以及疫苗临床试验和血清阳性率研究。保持结果之间的相关性。此外，当用登革热病毒样品测试时，其交叉反应性比MAC-ELISA低50.53％。该荧光中和试验可用于ZIKV感染的临床诊断确认，以及疫苗临床试验和血清阳性率研究。